HIV Research for Prevention 2014 AIDS Vaccine, Microbicide and ARV-based Prevention Science

Transcription

1 HIV Research for Prevention 204 AIDS Vaccine, Microbicide and ARV-based Prevention Science Cape Town, South Africa 28 3 October 204 Cape Town, South Africa 28 3 October 204

2 HIV Research for Prevention 204: AIDS Vaccine, Microbicide and ARV-based Prevention Science PROGRAM-AT-A-GLANCE Tuesday, 28 October Wednesday, 29 October Thursday, 30 October Friday, 3 October Opening Plenary Session 0 08:5 0:30 State of the Art: Biomedical Prevention in 204 (PL0) Level 0, Auditorium 0 Plenary Session 02 08:30 0:00 Targeting Biomedical Preventions to Different At-risk Populations (PL02) Level 0, Auditorium 0 Plenary Session 03 08:30 0:00 Importance of Mucosa in Prevention Research (PL03) Level 0, Auditorium 0 Refreshment Break 0:30 :00, Level 0, Ballroom Foyer Poster Session 0 I Refreshment Break 0:00 :00, Level 0, Hall 2 Poster Session 02 I Refreshment Break 0:00 :00, Level 0, Hall 2 Oral Abstract Sessions :00 2:30 B Cell Immunogen Design (OA) Level 0, Auditorium 0 Microbicides: Male Partner Engagement and Sexual Behaviors (OA2) Level 0, Meeting Room.60 Animal Model Studies of Microbicides and Injectables (OA3) Level 0, Auditorium 02 Innate Immunity (OA4) Level 02, Meeting Room 2.40 Vaccine, Viral Latency and Cure (OA5) Level 02, Roof Terrace Room Oral Abstract Sessions :00 2:30 Vaccine Development: Emerging Insights (OA) Level 0, Auditorium 0 Towards Broadly Neutralizing Antibody Induction (OA2) Level 0, Meeting Room.60 ARV Exposure and Efficacy in the Genital Tract (OA3) Level 0, Auditorium 02 Host Factors: Injury, Acquisition and Infection (OA4) Level 02, Meeting Room 2.40 PrEP and Microbicide Adherence in Women (OA5) Level 02, Roof Terrace Room Oral Abstract Sessions :00 2:30 Viral Transmission Studies (OA2) Level 0, Auditorium 0 Cell and Tissue Models of ARVs for Prevention (OA22) Level 0, Meeting Room.60 Pregnancy Intentions, Safe Conception and PMTCT (OA23) Level 0, Auditorium 02 Mucosal Responses (OA24) Level 02, Meeting Room 2.40 Adjuvants and Immunogens (OA25) Level 02, Roof Terrace Room Delegate and Networking Lunch 2:30 3:30, Level 0, Ballroom E+W Delegate and Networking Lunch 2:30 3:30, Level 0, Hall 2 Delegate and Networking Lunch 2:30 3:30, Level 0, Hall 2 Oral Abstract Sessions 3:30 5:00 B Cell Repertoires for Protection (OA6) Level 0, Auditorium 0 Risk and Prevention for Men who Have Sex with Men (OA7) Level 0, Meeting Room.60 Correlates of Protection and Exposure (OA8) Level 0, Auditorium 02 Engaging, Recruiting and Retaining Trial Participants (OA9) Level 02, Meeting Room 2.40 Bacterial Vaginosis and HSV-2: Impact on Genital Immunity (OA0) Level 02, Roof Terrace Room Oral Abstract Sessions 3:30 5:00 Novel Vaccine Concepts (OA6) Level 0, Auditorium 0 Mucosal Target and Effector Cells (OA7) Level 0, Meeting Room.60 Evaluation of Novel Biomedical Interventions (OA8) Level 0, Auditorium 02 Good Participatory Practices in HIV Prevention (OA9) Level 02, Meeting Room 2.40 Reproductive Hormones and HIV Risk (OA20) Level 02, Roof Terrace Room Oral Abstract Sessions 3:30 5:00 Microbicides and Multipurpose Prevention Technologies (OA26) Level 0, Auditorium 0 PrEP: Self-testing, Safety and Modeling (OA27) Level 0, Meeting Room.60 Treating and Preventing: the Role of ARVs (OA28) Level 0, Auditorium 02 T Cell Immunity (OA29) Level 02, Meeting Room 2.40 Antibody Functions and Protection (OA30) Level 02, Roof Terrace Room Non-Abstract Driven Sessions 08:30 0:00 Multipurpose Technologies (RT5) Level 0, Meeting Room.60 Challenges of Biomedical HIV Prevention Trials (SY0) Level 0, Auditorium 02 Emerging Areas in Immunity (SY) Level 02, Meeting Room 2.40 Differential Use of Antibodies in Prevention (SY2) Level 02, Roof Terrace Room Refreshment Break 0:00 0:30, Auditorium 0 Foyer Closing Plenary Session 04 0:30 2:30 Mind the Gap: Bridging from Trial Success to Access (PL04) Level 0, Auditorium 0 Afternoon Satellites 3:30 7:30 For Satellite Sessions on Monday, 27 October and Friday 3 October, visit satellite-sessions to view the program. Refreshment Break 5:00 5:30, Level 0, Ballroom Foyer Refreshment Break 5:00 5:30, Level 0, Hall 2 Refreshment Break 5:00 5:30, Level 0, Hall 2 Non-Abstract Driven Sessions 5:30 7:00 Overcoming Barriers to Broadly Neutralizing Antibody Induction (SY) Level 0, Meeting Room.60 Advances in Animal Models (SY2) Level 0, Auditorium 02 Treatment as Prevention: The Promise and the Perils (SY3) Level 02, Meeting Room 2.40 Building Combination Prevention Trials (SY4) Level 02, Roof Terrace Room Welcome Reception 7:00 9:00 Level 0, Ballroom E+W Please visit hivr4p.org/program/posters to view poster presentations by theme. Cape Town, South Africa Non-Abstract Driven Sessions 5:30 7:00 The Role of Structure Based Design in Vaccine Development and Immunoprophylaxis (SY5) Level 0, Auditorium 0 Harnessing Antibody Effector Functions (SY6) Level 0, Meeting Room.60 Novel Formulations and Sustained Delivery of Antiretrovirals (SY7) Level 0, Auditorium 02 Safer Sex in 204: Has the Paradigm Shifted? (RT) Level 02, Meeting Room 2.40 Prevention of Mother-to-Child Transmission Revisited (RT2) Level 02, Roof Terrace Room Poster Session 0 and Reception 7:00 8:30 Level 0, Hall 2 Oral Poster Discussions, 7:5 8:05 Community Engagement and Advocacy (PD) Room A Correlates of Protection in Highly Exposed Seronegative People (PD2) Room B Preclinical and Clinical Vaccine Trials (PD3) Room C Non-Abstract Driven Sessions 5:30 7:00 Diffusion of Innovation: Accelerating Along the Research to Rollout Continuum (RT3) Level 0, Meeting Room.60 Research Where and With Whom it Matters (RT4) Level 0, Auditorium 02 Mucosal Barriers to Infection (SY8) Level 02, Meeting Room 2.40 Sustaining Durability of Responses (SY9) Level 02, Roof Terrace Room Poster Session 02 and Reception 7:00 8:30 Level 0, Hall 2 Oral Poster Discussions, 7:5 8:05 Behavioral and Social Sciences (PD4) Room A Glycans and Antibody Effector Functions (PD5) Room B Policy, Advocacy and Modeling (PD6) Room C Key PL Plenary Session OA Oral Abstract Session SY Symposium RT Roundtable PD Poster Discussion

3 ABSTRACT BOOK 28 3 October 204 Cape Town, South Africa Abstracts will be published as an online supplement at at the conclusion of the conference on Friday, 3 October.

4 HIV Research for Prevention 204 HIV R4P

5 Contents Program at a Glance...Inside Front Cover Conference Committees Award and Scholarship Recipients Presentations Plenary Sessions Non-Abstract Driven Sessions Oral Abstract Sessions: Tuesday, 28 October Oral Abstract Sessions: Wednesday, 29 October Oral Abstract Sessions: Thursday, 30 October Poster Discussions: Wednesday, 29 October Poster Discussions: Thursday, 30 October Overview of Poster Sessions Posters: Wednesday, 29 October Posters: Thursday, 30 October Author Index Notes Certificate of Attendance Partner Recognition... Inside Back Cover Late Breaker Abstract... LB Level... L New Investigator Awardee.. NIA Oral Abstract.... OA Plenary Session.... PL Abbreviations Poster.... P Poster Discussion.... PD Roundtable.... RT Symposium Session.... SY CHICAGO Satellites begin on Monday, 7 October Tuesday, 8 October Friday, 2 October 5

6 Conference Committees CONFERENCE COMMITTEES CONFERENCE CHAIRS Sharon Hillier University of Pittsburgh, United States Eric Hunter Emory University, Atlanta, United States Anatoli Kamali Medical Research Council/UVRI Uganda Research Unit on AIDS, Entebbe, Uganda Helen Rees Wits Reproductive Health and HIV Institute (Wits RHI), Johannesburg, South Africa Robin Shattock Imperial College, London, United Kingdom PROGRAM ORGANIZING COMMITTEE MEMBERS Alash le Abimiku Institute of Human Virology, University of Maryland School of Medicine, United States Susan Barnett Novartis, United States Dan Barouch Beth Israel Deaconess Medical Center, Ragon Institute of MGH MIT and Harvard, United States Stephen Becker The Bill & Melinda Gates Foundation, United States Gina Brown Office of AIDS Research, National Institutes of Health, United States Susan Buchbinder San Francisco Department of Public Health, United States Elizabeth Bukusi Kenya Medical Research Institute, Kenya Dennis Burton The Scripps Research Institute, United States Nomita Chandhiok Indian Council of Medical Research, India Z. Mike Chirenje University of Zimbabwe - University of California Collaborative Research Programme, Zimbabwe Myron Cohen University of North Carolina, United States David Cooper Kirby Institute, University of New South Wales, Australia Carl Dieffenbach National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Patricia Fast International AIDS Vaccine Initiative, United States Glenda Gray Perinatal HIV Research Unit and South African Medical Research Council, South Africa Scott Hammer Columbia University, United States Catherine Hankins Amsterdam Institute for Global Health and Development, Netherlands Angela Kashuba University of North Carolina, United States Milly Katana John Snow, Inc., Uganda Roger Le Grand Commissariat à l Énergie Atomique, France Bonnie Mathieson Office of AIDS Research, National Institutes of Health, United States Andrew McMichael Oxford University, United Kingdom Lynn Morris National Institute for Communicable Diseases, South Africa Thumbi Ndung u University of KwaZulu-Natal, South Africa Julie Overbaugh Fred Hutchinson Cancer Research Center, United States Jim Pickett International Rectal Microbicide Advocates, United States Punnee Pitisuttithum Vaccine Trial Centre, Mahidol University, Thailand Gita Ramjee HIV Prevention Research Unit, South African Medical Research Council, South Africa Merlin Robb US Military HIV Research Program, United States Nina Russell The Bill & Melinda Gates Foundation, United States Barbara Shacklett University of California at Davis, United States Yiming Shao National Center for AIDS/STD Control and Prevention (NCAIDS), Department of the Research on Virology and Immunology, and Chinese Center fo Disease Control and Prevention, China Bill Snow Global HIV Vaccine Enterprise, United States Morenike Ukpong Obafemi Awolowo University; HIV Vaccine and Microbicide Advocacy Society, Nigeria Mitchell Warren AVAC: Global Advocacy for HIV Prevention, United States Carolyn Williamson University of Cape Town, South Africa Ntando Yola Desmond Tutu HIV Foundation, South Africa 6 HIV Research for Prevention 204 HIV R4P

7 Conference Committees ABSTRACT REVIEW COMMITTEE In addition to the Program Organizing Committee, HIV R4P would like to thank the following individuals for reviewing abstracts. Todd Allen Ragon Institute of MGH, MIT and Harvard, United States Galit Alter Ragon Institute of MGH, MIT and Harvard, United States Marcus Altfeld Heinrich Pette Institute, Germany Rama Amara The Yerkes National Primate Research Center, Emory University, United States Omu Anzala KAVI Institute of Clinical Research, University of Nairobi, Kenya Jared Baeten University of Washington, United States Prince Bahati International AIDS Vaccine Initiative, Kenya Christian Brander IrsiCaixa Institute for AIDS Research, HIVACAT, Spain Robert Chen Centers for Disease Control and Prevention, United States Josephine Cox International AIDS Vaccine Initiative, United States Cynthia Derdeyn Emory University, United States Charlene Dezzutti University of Pittsburgh, United States Gustavo Doncel CONRAD/Eastern Virginia Medical School, United States Daniel Douek National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Keith Fowke University of Manitoba, Canada Nicole Frahm Fred Hutchinson Cancer Research Center, United States David Friend CONRAD, United States Henry Gabelnick Independent Consultant, United States J. Victor Garcia University of North Carolina at Chapel Hill, United States Paul Goepfert University of Alabama at Birmingham, United States Barney Graham Vaccine Research Center, National Insittute of Allergy and Infectious Dieases, National Institutes of Health, United States Tomas Hanke University of Oxford, United Kingdom Barton Haynes Duke Human Vaccine Institute, Duke University, United States Craig Hendrix Johns Hopkins University, United States Walid Heneine Centers for Disease Control and Prevention, United States Betsy Herold Albert Einstein College of Medicine, United States Saidi Kapiga London School of Hygiene and Tropical Medicine, Mwanza Intervention Trials Unit, United Kingdom Stephen Kent University of Melbourne, Australia Jerome Kim US Military HIV Research Program, United States Wayne Koff International AIDS Vaccine Initiative, United States Charles Lacey University of York, United Kingdom Yves Levy Vaccine Research Institute, France George Lewis Institute of Human Virology, University of Maryland School of Medicine, United States Amapola Manrique Global HIV Vaccine Enterprise, United States Jeanne Marrazzo University of Washington, United States John Mascola National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Benoit Masse CHU Sainte Justine and the Montreal Children s Hospital, Canada Kenneth Mayer Fenway Health, United States Barbara Mensch Population Council, United States Nelson Michael US Military HIV Research Program, United States Penny Moore National Institute for Communicable Diseases, South Africa Jeremy Nuttall International Partnership for Microbicides, United States Jean Patterson Office of AIDS Research, National Institutes of Health, United States Dorothy Patton University of Washington, United States Damian Purcell University of Melbourne, Australia Harriet Robinson Geovax Inc., United States Lisa Rohan Department of Pharmaceutical Sciences School of Pharmacy, University of Pittsburgh, Magee-Women s Research Institute, United States Morgane Rolland US Military HIV Research Program, Henry M. Jackson Foundation, United States Joseph Romano NWJ Group LLC, United States Jeffrey Safrit Elizabeth Glaser Pediatric AIDS Foundation, United States Eric Sandström Karolinska Institutet, Sweden William Schief The Scripps Research Institute, International AIDS Vaccine Initiative, Ragon Institute of MGH, MIT and Harvard, United States Alexandra Schuetz Armed Forces Research Institute for Medical Science, United States Olivier Schwartz Institut Pasteur, France Guido Silvestri Emory University, United States Jonathan Stadler Wits Reproductive Health and HIV Institute, South Africa Alan Stone MEDSA Ltd., United Kingdom CONFERENCE COMMITTEES 7

8 Conference Committees CONFERENCE COMMITTEES ABSTRACT REVIEW COMMITTEE (continued) Jim Tartaglia Sanofi Pasteur, United States Caroline Tiemessen National Institute for Communicable Diseases, South Africa Georgia Tomaras Duke Human Vaccine Institute, Duke University, United States Alexandra Trkola University of Zurich, Germany Jim Turpin National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States SECRETARIAT Mark Aurigemma Communications Consultant, United States Jennifer Brunet Global HIV Vaccine Enterprise, United States Karlyanna Kopra Conference Solutions, United States Amapola Manrique Global HIV Vaccine Enterprise, United States Lut Van Damme The Bill & Melinda Gates Foundation, United States Ronald Veazey Tulane National Primate Research Center, United States Steven Wakefield HIV Vaccine Trials Network, United States Anna-Lise Williamson University of Cape Town, South Africa Allen Zhiwei Wu Nanjing University, China Kristin Morrell Global HIV Vaccine Enterprise, United States Kate Porter Conference Solutions, United States Nicole Santamaria Conference Solutions, United States 8 HIV Research for Prevention 204 HIV R4P

9 Scholars NEW INVESTIGATOR AWARDEES HIV R4P 204 is proud to introduce this year s New Investigator Awardees. Siriwat Akapirat Armed Forces Research Institute of Medical Sciences, Department of Retrovirology, Thailand Jinal Bhiman National Institute for Communicable Diseases and the University of the Witwatersrand, South Africa Elizabeth Byrne Harvard University, United States Minlu Hu Magee-Womens Research Institute / University of Pittsburgh, United States Luca Schifanella National Cancer Institute, National Institutes of Health, United States CONFERENCE SCHOLARS Scholarships were made possible by generous donations from our partners and from registration funds. Andrew Abaasa Medical Research Council / Uganda Virus Research Institute, Uganda Nathlee Abbai South African Medical Research Council, HIV Prevention Research Unit, South Africa Olatunji Adetokunboh Stellenbosch University, South Africa Wbeimar Aguilar Grupo Inmunovirología, Facultad de Medicina, Universidad de Antioquia, Colombia Nurelign Ahmed Projet San Francisco, Rwanda-Zambia HIV Research Group, Rwanda Seema Ajbani National Institute for Research in Reproductive Health, India Carolyne Akello Makerere University - Johns Hopkins University Research Collaboration, Uganda Aderemi Oyekola Alagbe Namibia Ministry of Health and Social Services, Namibia Muriel Aldunate Burnet Institute for Medical Research / Monash University, Australia Mallika Alexander National AIDS Research Institute, India Kabamba Alexandre Council for Scientific and Industrial Research, South Africa Mambo Amisi Modeste Humanitarian Action for Health and Community Development, Democratic Republic of the Congo Obiajulu Amuamuziam New HIV Vaccine and Microbicide Advocacy Society, Nigeria Winnie Apidi University of Manitoba, Canada Eva Archer Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Gershim Asiki Medical Research Council / Uganda Virus Research Institute, Uganda Abdul Azeem National AIDS Control Program, Pakistan Veenu Bala Central Drug Research Institute, India Alejandro Balazs Ragon Institute of MGH, MIT and Harvard, United States Cameron Ball University of Washington, United States Shaun Barnabas University of Cape Town, South Africa Deborah Baron Wits Reproductive Health and HIV Institute, South Africa Tahir Bashir Dar National Institute for Research in Reproductive Health, India Cheryl Baxter Centre for AIDS Programme of Research in South Africa, South Africa Mara Biasin University of Milan, Italy Patrick Bitangumutwenzi Young Women s Knowledge and Leadership Institute, Burundi Saikat Boliar Translational Health Science and Technology Institute, India Jacqui Brener Peter Medewar Building for Pathogen Research, University of Oxford, United Kingdom Charles Brown Infectious Diseases Institute, AVAC, Uganda William Brown, III HIV Center for Clinical and Behavioral Studies, Columbia University, United States Omkar Chaudhary All India Institute of Medical Sciences, India Hannah Cheeseman Imperial College London, United Kingdom Yan Li Chen National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, China Christopher Chianese Fenway Health, United States Clever Chilende Treatment Advocacy and Literacy Campaign, Zambia Jonah Chinga Gay and Lesbian Coalition of Kenya, Kenya Bhavna Chohan Kenya Medical Research Institute, Kenya Amy Chung Ragon Institute of MGH, MIT and Harvard, United States Daniel Claiborne Emory University, United States Núria Climent AIDS Research Group, CELLEX-IDIBAPS-HIVACAT, Hospital Clinic i Provincial de Barcelona, Spain Sarah Cohen Wits Reproductive Health and HIV Institute, South Africa Smritee Dabee Institute of Infectious Disease and Molecular Medicine, University of Cape Town, South Africa Christine Dahlke Universitätsklinikum Hamburg-Eppendorf, Germany AWARD / SCHOLARS 9

10 Scholars SCHOLARS CONFERENCE SCHOLARS (continued) José das Neves Instituto de Engenharia Biomédica, Porto, Portugal, Portugal Martin Deymier Emory University, United States Kathleen Doherty Vanderbilt University, Ragon Institute of MGH, MIT and Harvard, United States Clare Dott Wits Reproductive Health and HIV Institute, South Africa Victor Yimin Du University of Alabama at Birmingham, United States Zoe Duby University of Cape Town, Desmond Tutu HIV Foundation, South Africa Juliane Etima Makerere University - Johns Hopkins University Research Collaboration, Uganda Abbey Evans Imperial College London, United Kingdom Yu Feng International AIDS Vaccine Initiative, United States Susan Fetherston University of Ulster, United Kingdom Jacqueline Flynn Burnet Institute, Australia Pascaline Fonteh University of Pretoria, South Africa Alemju Fontu Association Camerouniase Pour le Marketing Social, Cameroon Anna Forbes Independent Consultant, United States Joseph Francica National Institutes of Health, United States Ereshia Gabier South African National Bioinformatics Institute / University of the Western Cape, South Africa Tanuja Gengiah Centre for AIDS Programme of Research in South Africa, South Africa Yanina Ghiglione Instituto de Investigaciones Biomédicas en Retrovirus y Sida, Argentina Anna Gibbs Karolinska Institutet, Sweden Wesley Grimm Northwestern University, United States Anneke Grobler Centre for AIDS Programme of Research in South Africa, South Africa Tiffany Grooms-Williams University of Louisville, United States Ntombesizwe Nombasa Gxuluwe World AIDS Campaign, South Africa Antje Heit Fred Hutchinson Cancer Research Center, United States Tiffany Hensley-McBain University of Washington, United States Carolina Herrera Imperial College London, United Kingdom Heather Hong Centre for HIV and STIs, National Institute for Communicable Diseases of the National Health Laboratory Service, and the Faculty of Health Sciences, University of the Witwatersrand, South Africa Chiedu Ifekandu Population Council, Nigeria Rosine Ingabire Projet San Francisco, Rwanda-Zambia HIV Research Group, Rwanda Desmond Iriaye Population Council, Nigeria Lakshmi Jagesur South African Medical Research Council, South Africa Mariel Jais Milken Institute School of Public Health, The George Washington University, United States Kara Jensen University of North Carolina and Center for AIDS Research, Chapel Hill, United States Sanne Skov Jensen Statens Serum Institut, Denmark Vineet Joag University of Toronto, Canada Tsungai Ivai Jongwe University of Cape Town, South Africa Kadryn Kadasia Boston University, United States Betty Kamira Makerere University - Johns Hopkins University Research Collaboration, Uganda Brian Kanyemba Desmond Tutu HIV Foundation, South Africa William Kilembe Rwanda Zambia HIV Research Group, Zambia Linda Kimaru Rwanda Zambia HIV Research Group, United States Janine Kimpel Innsbruck Medical University, Austria Deborah King Imperial College London, United Kingdom Rose Kitawi Centre for Research in Therapeutic Sciences, Kenya Nichole Klatt University of Washington, United States Henrik Kløverpris KwaZulu-Natal Research Institute for Tuberculosis and HIV, South Africa Rewa Kohli National AIDS Research Institute, India Catherine Kegakilwe Koofhethile HIV Pathogenesis Programme, South Africa Jennifer Kotlewski Rwanda Zambia HIV Research Group, Zambia Jean-Mari Kriek Institute of Infectious Disease and Molecular Medicine, University of Cape Town, South Africa Sylvia Kusemererwa Medical Research Council / Uganda Virus Research Institute, Uganda Zachary Kwena Kenya Medical Research Institute, Kenya Rachel Kyeyune Infectious Diseases Institute, Uganda Jordan Kyongo Institute of Tropical Medicine - Antwerp, Kenya Faatima Laher HIV Pathogenesis Programme, South Africa Elise Landais International AIDS Vaccine Initiative, United States 0 HIV Research for Prevention 204 HIV R4P

11 Scholars CONFERENCE SCHOLARS (continued) Robert Langat KAVI-Institute of Clinical Research,University of Nairobi, Kenya Ria Lassauniere National Institute for Communicable Diseases, South Africa Marc-André LeBlanc Self-employed, Canada Boon Kiat Lee AIDS Institute, The University of Hong Kong, Hong Kong Duduzile Lembethe MatCH Research University of Witwatersrand, South Africa Maria Lemos Fred Hutchinson Cancer Research Center, United States Mildie Leuvennink International Partnership for Microbicides, South Africa Clement Levin INSERM U35 CIMI, Paris, France Haiying Li Beth Israel Deaconess Medical Center, Harvard Medical School, United States Lenine Liebenberg Centre for AIDS Programme of Research in South Africa, South Africa Namal P.M. Liyanage National Cancer Institute, National Institutes of Health, United States Murray Logan University of Cape Town, South Africa Evelyn Lumngwena University of Cape Town, South Africa Alison Mahan Ragon Institute of MGH, MIT and Harvard, United States Mookho Malahleha Setshaba Research Centre, South Africa Thandi Maluka SHIPP, South Africa Jai Marathe Boston University, United States Elena Martinelli Population Council, United States Enrique Martin-Gayo Ragon Institute of MGH, MIT and Harvard, United States Yasuhiro Maruta Matsushita Project Laboratory, Center for AIDS Research, Kumamoto University, Kumamoto, Japan Rose Masilo Madibeng Centre for Research, South Africa Flavia Matovu Kiweewa Makerere University - Johns Hopkins University Research Collaboration, Uganda Elizabeth Mbabazi Atuhurra Medical Research Council, Uganda Andrew McGuire Seattle Biomed, United States Eric Mcheka National Association for People Living with HIV/AIDS in Malawi, Malawi Lyle McKinnon Centre for AIDS Programme of Research in South Africa, South Africa Meron Mengistu Institute of Human Virology, United States Sergey Menis The Scripps Research Institute, United States Melanie Merbah U.S. Military HIV Research Program, Henry M. Jackson Foundation, United States Clint Mercuur South African National Bioinformatics Institute, South Africa Nyaradzo Mgodi University of Zimbabwe - University of California Collaborative Research Programme, Zimbabwe Maureen Milanga AIDS Law Project, Kenya Lori Miller London School of Hygiene & Tropical Medicine, United Kingdom Brenda Gati Mirembe Makerere University - Johns Hopkins University Research Collaboration, Uganda Nonhlanhla Mkhize National Institute for Communicable Diseases, South Africa Kathryn Mngadi Centre for AIDS Programme of Research in South Africa, South Africa Daniela Monaco Emory University, United States Jayajothi Moodley South African Medical Research Council, HIV Prevention Research Unit, South Africa Jeeva Moodley South African Medical Research Council, HIV Prevention Research Unit, South Africa Nishila Moodley Perinatal HIV Research Unit, South Africa Neetha Morar South African Medical Research Council, HIV Prevention Research Unit, South Africa Sara Morón-López IrsiCaixa AIDS Research Institute - HIVACAT, Spain Beatriz Mothe IrsiCaixa AIDS Research Institute - HIVACAT, Spain Thandeka Moyo University of Cape Town, South Africa Juliet Mpendo Uganda Virus Research Institute, International AIDS Vaccine Institute, Uganda Sandra Mudhune Wits Reproductive Health and HIV Institute, South Africa Peter Mugo Kenya Medical Research Institute, Wellcome Trust, Kenya Kenneth Mugwanya University of Washington, United States Andrew Mujugira University of Washington, Uganda Wendy Murillo Universidad Nacional Autonoma de Honduras, Honduras Eva Muro Kilimanjaro Clinical Research Institute, United Republic of Tanzania Petina Musara University of Zimbabwe - University of California San Francisco Collaborative Research Programme, Zimbabwe Leonard Mutisya The East African Sexual Health and Right Initiative, Kenya Shem Mutuiri Institute of Primate Research, Kenya Rosemary Muwawu Makerere University - Johns Hopkins University Research Collaboration, Uganda SCHOLARS

12 Scholars SCHOLARS CONFERENCE SCHOLARS (continued) Simon Mwangi Bar Hostess Empowerment and Support Program, Kenya Lawrence Mwihaki Partners in Prevention Thika - University of Washington Site, Kenya Philip Mwimanzi Simon Fraser University, Canada Pumeza Mzizi Perinatal HIV Research Unit, Wits Health Consortium, South Africa Sarita Naidoo South African Medical Research Council, HIV Prevention Research Unit, South Africa Lillian Naigaga Mutengu International AIDS Vaccine Initiative, Kenya Teopista Nakyanzi Makerere University - Johns Hopkins University Research Collaboration, Uganda Sophie Clare Nanziri Makerere University - John Hopkins University Research Collaboration, Uganda Tashini Nayager South African Medical Research Council, HIV Prevention Unit, South Africa Pepukai Ndadziyira University of Zimbabwe - University of California San Francisco Collaborative Research Programme, Zimbabwe Zaza Ndhlovu University of KwaZulu Natal, Nelson Mandela School of Medicine, South Africa Bongiwe Ndlovu HIV Pathogenesis Programme, South Africa Evanson Ndung u Kenya Medical Research Institute, Kenya Duduzile Ndwandwe South African Medical Research Council, HIV Prevention Research Unit, South Africa Robert Newells AVAC PxROAR, United States Kenneth Ngure Jomo Kenyatta University of Agriculture and Technology - College of Health Sciences, Kenya Nadesh Ngechae Nji Chantal Biya International Reference Center for Research on the Prevention and Management of HIV/AIDS, Cameroon Stella Njuguna Kenya Medical Research Institute, Kenya Chanda Nsofwa Family Health Trust, Zambia Fanelesibonge Ntombela Centre for AIDS Programme of Research in South Africa, South Africa Julien Nyombayire Projet San Francisco, Rwanda-Zambia HIV Research Group, Rwanda Ayodeji Oginni Population Council, Nigeria Sandra Okala Imperial College London, United Kingdom Dismas Oketch Moi University School of Medicine, Kenya Kennedy Olango Men Against AIDS Youth Group, Kenya Catherine Oldenburg Harvard School of Public Health, United States Alex Olvera IrsiCaixa AIDS Research Institute - HIVACAT, Spain Shatha Omar University of Cape Town, South Africa Everlyne Ombati Kenya Medical Research Institute, Kenya Gloria Omosa-Manyonyi University of Nairobi, Kenya Simon Ondiek HIV/AIDS Research & Advocacy Program, Kenya Sylvia Onyango Medical Research Council / Uganda Virus Research Institute, Uganda Chiara Orlandi Institute of Human Virology - University of Maryland, United States Sophia Osawe Institute of Human Virology, Nigeria George Victor Owino International AIDS Vaccine Initiative, Kenya Patrick Owiti Kenya Medical Research Institute / Family AIDS Care and Education Services project, Kenya Mickey Patel Geisel School of Medicine at Dartmouth, United States Arendevi Pather South African Medical Research Council, HIV Prevention Research Unit, South Africa Shilpa Patil Translational Health Science and Technology Institute, India Sinazo Pato Wits Reproductive Health and HIV Institute, South Africa Laura Pattacini Fred Hutchinson Cancer Research Center, United States San Patten San Patten and Associates, Inc., Canada Binghao Peng University of Alabama at Birmingham, United States Catia Perciani University of Toronto, Canada Capucine Phelip Institut de Biologie et Chimie des Protéines, LBTI, UMR CNRS/Université de Lyon, France Jessica Phillip HIV Prevention Research Unit, Medical Research Council, South Africa Anabela Picton Centre for HIV and STIs, National Institute for Communicable Diseases of the National Health Laboratory Service, and the Faculty of Health Sciences, University of the Witwatersrand, South Africa Anna Piddubna Sumy State University, Ukraine Diantha Pillay University of KwaZulu-Natal, Centre for AIDS Programme of Research in South Africa, South Africa Anastasia Pokrovskaya Federal Budget Institution of Science Central Scientific Research Institute of Epidemiology of Rospotrebnadzor, Russian Federal AIDS Centre, Russian Federation Gloria Preza University of Southern California, United States Jessica Prince Emory University, United States Zakiya Qualls Rutgers Biomedical and Health Sciences, United States 2 HIV Research for Prevention 204 HIV R4P

13 Scholars CONFERENCE SCHOLARS (continued) Pranitha Ramchuran Wits Reproductive Health and HIV Institute, South Africa Srinika Ranasinghe Ragon Institute of MGH, MIT and Harvard, United States Mary-Jane Ratlhagana International Training and Education for Health, South Africa Krishnaveni Reddy Wits Reproductive Health and HIV Institute (Wits RHI), South Africa KVR Reddy National Institute for Research in Reproductive Health, India Simone Richardson Centre for HIV and STIs, National Institute for Communicable Diseases of the National Health Laboratory Service, and the Faculty of Health Sciences, University of the Witwatersrand, South Africa Laura Richert Spuhler University of Washington, United States Meika Richmond University of Manitoba, Canada Marta Rodriguez Garcia Geisel School of Medicine at Dartmouth, United States Maria Julia Ruiz Instituto de Investigaciones Biomédicas en Retrovirus y Sida, Universidad de Buenos Aires/CONICET, Argentina Wiriya Rutvisuttinunt AFRIMS, Thailand Richard Rwanyonga Medical Research Council / Uganda Virus Research Institute, Uganda Darpun Sachdev Bridge HIV, San Francisco Department of Public Health and Center for AIDS Prevention Studies, University of California San Francisco, United States Anwesha Sanyal University of Pittsburgh Graduate School of Public Health, United States Irma Saulle University of Milan, Italy Cathrine Scheepers Center for HIV and STIs, National Institute for Communicable Diseases and the University of the Witwatersrand, South Africa Torben Schiffner University of Oxford, United Kingdom Veronika Schmid Institute of Medical Microbiology and Hygiene, University of Regensburg, Germany Jeffrey Schneider Northwestern University, United States Yanille Scott University of Pittsburgh Graduate School of Public Health, United States Adekemi Sekoni University of Lagos, Nigeria Kwame Shanaube ZAMBART, Zambia Remmy Shawa Sonke Gender Justice, South Africa Mahesh Sherkar Shri Vivekanand Nursing Home Trust s College of Pharmacy, Rahuri, Ahmednagar, Maharashtra, India, India Stuart Sievers California Institute of Technology, United States Paola Silveira Federal University of Rio de Janeiro, Brazil Chitra Singh Centre for AIDS Programme of Research in South Africa, South Africa Satya Prakash Singh Indian Institute of Technology - Madras, India Udaya Pratap Singh Sam Higginbottom Institute of Agriculture, Technology & Sciences, India Magdalena Sips Ragon Institute of MGH, MIT and Harvard, United States Tsakani Sithole AURUM Institute, South Africa Aida Sivro University of Manitoba, Canada Kennedy Smart Institute of Human Virology, Nigeria Kieron Smith University of Aberdeen, United Kingdom Melissa Smith University of Manitoba, Canada Stacey Smith Emory University, Yerkes National Primate Research Center, United States Olivia Snyder University of North Carolina, Chapel Hill, United States Devin Sok The Scripps Research Institute, United States Livingstone Ssali The AIDS Support Organisation, TASO, Uganda Derek Stein University of Manitoba, Canada Renee Street South African Medical Research Council, HIV Prevention Research Unit, South Africa Bin Su INSERM U09, FMTS, Université de Strasbourg, France Ibrahim Suleiman Population Council, Nigeria Zehua Sun AIDS Institute, The University of Hong Kong, Hong Kong Nsubuga Supercharger Moses Joint Clinical Research Centre, Uganda Niwat Suwanphatthana Thai NGO Coalition on AIDS, Thailand Jessica Terlikowski AIDS Foundation of Chicago, United States Sydney Tetteh Hushie Global Youth Coalition on HIV and AIDS, Ghana Aime Marcel Tongo Passo International Centre for Genetic Engineering and Biotechnology, University of Cape Town, South Africa Hung Trinh U.S. Military HIV Research Program, United States Damien Tully Ragon Institute of MGH, MIT and Harvard, United States Morenike Ukpong Institute of Public Health, Obafemi Awolowo University; New HIV Vaccine and Microbicide Advocacy Society, Nigeria DaShawn Usher Project ACHIEVE, United States Neliëtte Van Niekerk International Partnership for Microbicides, South Africa Thomas Vazquez I3 Laboratory - INSERM U959 - GHPS Paris, France SCHOLARS 3

14 Scholars CONFERENCE SCHOLARS (continued) Bellington Vwalika Zambia Emory HIV Research Project, Zambia Dana Watnick Albert Einstein College of Medicine, United States Huamian Wei State Key Laboratory for Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, China Dawn Renee Weinman University of Pittsburgh, United States Tendayi Westerhof Women s Health HIV and AIDS Southern Africa, Zimbabwe Lee Adam Wheeler Harvard Medical School, United States Constantinos Kurt Wibmer Centre for HIV and STIs, National Institute for Communicable Diseases of the National Health Laboratory Service, and the Faculty of Health Sciences, University of the Witwatersrand, South Africa Wendy Winnall The University of Melbourne, Australia Xilin Wu The University of Hong Kong, Hong Kong Wildeman Zapata Universidad de Antioquia, Colombia Susanne Ziegler Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Germany Michael Zulu Doris Duke Medical Research Institute, HIV Pathogenesis Programme, University of KwaZulu-Natal, South Africa SCHOLARS 4 HIV Research for Prevention 204 HIV R4P

15 Opening Plenary: State of the Art Biomedical Prevention in 204 Tuesday, 28 October PL0.0 Prospects for an Antibody-based HIV Vaccine Lynn Morris,2 National Institute for Communicable Diseases of the NHLS, Centre for HIV & STI s: HIV Virology, Johannesburg, South Africa, 2 University of the Witwatersrand, Johannesburg, South Africa PL0.02 Advances in Antiretroviral-Based Prevention Research Jared Baeten University of Washington, Seattle, WA, United States Intensive efforts to develop a preventive HIV vaccine have been significantly bolstered by recent discoveries of broad and potent neutralizing antibodies in some HIV infected humans. Many of these antibodies have unusual genetic features that present a significant challenge for conventional vaccine approaches. Nevertheless, longitudinal studies are revealing how such lineages evolve and enabling the identification of germline B cells that an HIV vaccine would need to stimulate. Parallel viral genetic analysis further demonstrates how viral evolution shapes these responses. Collectively such studies in HIV- infected humans who develop broadly cross-neutralizing antibodies are providing important clues for HIV vaccine design. Antiretroviral-based HIV- prevention strategies including antiretroviral treatment (ART) to reduce the infectiousness of HIV- infected persons and oral, topical, and injectable antiretroviral preexposure prophylaxis (PrEP) for uninfected persons to prevent HIV- acquisition are groundbreaking new approaches for decreasing HIV- spread. The past three years have seen substantial advances in knowledge regarding ART and PrEP for HIV- prevention, including definitive demonstration in randomized trials that both ART and PrEP reduce HIV- risk and the development of normative guidance for prescribing these HIV- prevention strategies. Ongoing research into longer-acting PrEP agents could add new prevention options. There are numerous parallels in the opportunities ahead for ART and PrEP, including evaluating successful approaches to deliver these interventions to realize maximum population benefits, developing messaging that speaks to potential ART and PrEP users, understanding how sufficiently high adherence can be sustained to achieve high effectiveness, and integrating these strategies into health systems. Achieving success in antiretroviral-based prevention will demand the full force and breadth of prevention research and advocacy. PL0.03 Comprehensive HIV Prevention: Synergy Between Vaccine and Non-Vaccine Modalities Anthony S. Fauci National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States PLENARY SESSIONS 5

16 Wednesday, 29 October Plenary 02: Targeting Biomedical Preventions to Different At-Risk Populations PLENARY SESSIONS PL02.0 Tailoring Biomedical Preventive Interventions for Key Populations: Towards Safety, Efficacy, Effectiveness Chris Beyrer Johns Hopkins Bloomberg School of Public Health, Epidemiology, Baltimore, MD, United States PL02.02 Working with Special Populations within HIV Prevention Intervention Programs and Trials Bridget G. Haire,2 University of New South Wales, School of Public Health and Community Medicine, Sydney, Australia, 2 University of Sydney, Centre for Values, Ethics and the Law in Medicine, Sydney, Australia It is well accepted that effective HIV prevention must target the high risk populations in particular local epidemics, and that working collaboratively with these affected populations to design and deliver prevention interventions is an ethical requirement that can maximise uptake and acceptability. In 204, we have a greater than ever range of interventions to prevent HIV, but access, sustainability and adherence remain major issues, and there is no established optimal formula for implementing combination HIV prevention approaches in the specific population groups who need them most. In addition, some of the social and political barriers to effective implementation within key populations have become more entrenched. This poses particular problems both for ongoing research into new interventions and for programmatic implementation of proven interventions. This plenary will look at the evidence about the use of HIV prevention in particular populations, and consider how HIV prevention research and implementation can work within a broader human rights framework that takes into account social and political barriers that affect vulnerability, sustainability and adherence to prevention interventions. PL02.03 Tailoring Interventions to Different Populations Pontiano Kaleebu Basic Sciences, MRC/UVRI Uganda Research Unit on AIDS, Entebbe, Uganda UNAIDS estimates there were 6,300 new infections a day in 202, with 95% of these in low- and middle-income countries. Advances in biomedical HIV prevention are giving us hope that the HIV epidemic can be brought under control. Since Biomedical, Behavioural and Structural interventions intersect, it is important to understand and engage the different priority populations and communities in order to tailor interventions for maximum benefit. Social behavioural, health systems, community participation, epidemic stage, and policy are some factors that will determine the benefits for priority populations of the advances made in biomedical HIV prevention. For example, in most generalized epidemics effective prevention approaches are required for women who are at a disproportionate risk of HIV acquisition for biological as well as social reasons. Mobilising and meaningfully engaging key populations, such as people who inject drugs (PWID), men who have sex with men (MSM), sex workers (SW), and the fisherfolk (FF) in some countries will take different forms in local contexts for these often hard to reach populations. For example, in our studies in Uganda among SW, interventions are provided through special clinics, staffed by skilled health workers with knowledge and understanding of the sex work milieu, offer friendly services and involvement of peers in mobilisation and follow up. For both SW and FF, localising clinics and services nearer to their places of work is essential to their involvement. For FF this includes providing services that are open hours when they are not working. Examples of other tailored programmes engaging populations such as MSM and adolescents will be discussed. Investing resources and focusing national HIV prevention campaigns on well-defined strategies for and with populations that are key to the epidemic and key to the response will increase the potential for biomedical interventions to slow the HIV epidemic, reducing new infections. 6 HIV Research for Prevention 204 HIV R4P

17 Plenary 03: Importance of Mucosa in Prevention Research Thursday, 30 October PL03.0 Utilizing NHP Models to Understand Mucosal HIV Transmission and Dissemination Jake D. Estes Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, AIDS & Cancer Virus Program, Frederick, MD, United States PL03.02 Genital Inflammation and HIV Risk in Prevention Research Jo-Ann Passmore,2,3 University of Cape Town, Institute of Infectious Disease and Molecular Medicine, Cape Town, South Africa, 2 National Health Laboratory Service, Cape Town, South Africa, 3 CAPRISA, Durban, South Africa Over 80% of sexual HIV- transmissions originate from a single viral variant, but the underlying basis for this transmission bottleneck remains to be elucidated. Nonhuman primate models of mucosal virus transmission allow opportunities to gain insight into the basis of this mucosal bottleneck. This talk will focus on studies that utilize NHP models to understand i) the host innate antiviral responses during the earliest time points after mucosal SIV challenge and ii) the lymphatic drainage pathways of viral dissemination in order to better elucidate the earliest events of virus mucosal transmission and potential intervention strategies. PL03.03 Mucosal Immune Assays in HIV Vaccine Clinical Trials Omu A. Anzala KAVI - Institute of Clinical Research (KAVI-ICR), University of Nairobi, College of Health Sciences, Nairobi, Kenya Background: Mucosal immune responses and mucosal sampling from genitourinary (GU) and gastrointestinal (GI) tracts are at an increased focus for HIV vaccine research and development as well as other HIV prevention and treatment strategies that are targeted at mucosal surfaces. This is in realization that mucosal surface forms the major route of HIV acquisition and transmission across the world. Collection of mucosal samples during the conduct of clinical trials is associated with significant operational challenges, expenses, as well as some risk and discomfort to study participants. It is therefore critical that appropriate measures are taken into account including, (clinical, behavioral, and demographic characteristics) from study participants so that factors that may influence mucosal immunology and thus the interpretation of assay data are efficiently captured in parallel with mucosal specimens during the conduct of clinical trials. KAVI-Institute of Clinical Research at the University of Nairobi, and others have studied the acceptability and tolerability of repeated mucosal sampling in clinical trial participants as well as other participants at low risk of HIV infection. To this end the samples obtained were also used to establish and standardize the immune assays for adaptation in evaluation of mucosal immune responses in clinical trials. Discussion: Repeated mucosal sampling is achievable both in HIVinfected and in healthy adult HIV uninfected clinical trials participants. The sampling methods that have been studied include saliva, oral fluids, semen, cervical, vaginal, rectal, and gut. Participants consented to most specimen collection methods with the exception of rectal sampling. Samples obtained are of good quality for process, analysis and can be standardized for both T/B cell as well antibody assays for adaptation for use in evaluating mucosal immune responses in HIV vaccine clinical trials in addition to peripheral samples. PLENARY SESSIONS 7

18 Friday, 3 October Closing Plenary : Mind the Gap: Bridging from Trial Success to Access PL04.0 Translating Controlled Trial Results to the Cities & Villages of Africa: Past, Present, & Future Douglas N. Shaffer, Robert W. Eisinger, Deborah L. Birx Office of the U.S. Global AIDS Coordinator, U.S. Department of State, Washington, DC, United States PL04.02 Scaling-up HIV Prevention Science from the Laboratory to the Village Alex G. Coutinho Infectious Diseases Institute, Director, Kampala, Uganda HIV prevention science requires a critical mass scale-up to have a population impact. The talk will share experiences of scale-up with medical male circumcision and PMTCT and seek to learn lessons that can be utilized for emerging HIV prevention science. PL04.03 Antiretrovirals for Prevention Glenda Elisabeth Gray Department of Paediatrics, at the Perinatal HIV Research Unit, University of the Witwatersrand, Johannesburg, South Africa PLENARY SESSIONS 8 HIV Research for Prevention 204 HIV R4P

19 Tuesday, 28 October Symposium 0: Overcoming Barriers to Broadly Neutralizing Antibody Induction SY0.0 Prevention Strategies Based on HIV- Neutralizing Antibodies John Mascola National Institutes of Health, Vaccine Research Center, NIAID, Bethesda, MD, United States SY0.03 HIV Envelope Interactions with the Progenitor BCRs of Narrow and Broadly Neutralizing Antibodies Leonidas Stamatatos Seattle Biomedical Research Institute, Seattle, WA, United States Effective vaccines often generate protective antibody responses that are similar to those produced during natural infection. During HIV- infection, most individuals generate neutralizing antibodies that arise too late to be protective. However, during the course of infection, some individuals develop extraordinarily potent and broadly reactive neutralizing monoclonal antibodies (bnabs) that have been recently isolated and studied in detail. Such antibodies, when isolated and used for immunoprophylaxis in animal models, can completely block HIV infection. Thus, bnabs provided a template for the kind of immune response we aim to elicit via immunization. The analysis of their structural mode of recognition and how these antibodies develop to attain effective functional neutralization; i.e. genetic pathways of affinity maturation, can impact HIV vaccine design and prevention efforts in several ways including ) structure-based vaccine design, 2) understanding antibody evolution to optimize immunization strategies and 3) direct immunoprophylaxis by passive antibody or gene-based vectors. Highlights from each of these areas will be discussed. SY0.02 Broadly Neutralizing Antibody Responses in HIV- Infection: Insights from a Large Cohort and from Individuals Pascal Poignard IAVI Neutralizing Antibody Center at The Scripps Research Institute, La Jolla, CA, United States SY0.04 HIV- Mimicry of Host Antigens: Hiding in Plain Sight to Evade Immunity Garnett Kelsoe Duke University, Immunology and Human Vaccine Center, Durham, NC, United States Although controversial, it has been proposed that antibodies (Ab) that neutralize multiple HIV- clades (bnabs) are characteristically poly- or reactive to host antigens (autoreactive). We have identified a number of conserved mammalian autoantigens that are avidly bound by different bnabs, and have shown in mice that relaxation of immune tolerance significantly elevates Ab responses to HIV- neutralizing epitopes. The generation of knockin mice that express human bnab variable regions commonly has shown that the cross-recognition of host antigens is sufficient to block B-cell development or maturation by known tolerance mechanisms. Knockin strains that express non-neutralizing Ab variable regions support normal B-cell development. There is, however, no broad agreement as to whether HIV- mimicry of host determinants represents a crucial limitation of protective immunity to HIV- or is an ancillary property of Abs generated over the course of a chronic infection. I shall demonstrate that poly- and autoreactivity are characteristic of HIV- bnabs but not high affinity, strain-specific or non-neutralizing HIV- Abs generated in chronically infected individuals. As a class, bnabs are significantly more polyreactive and autoreactive than other HIV- Abs even though they are generated under similar conditions of chronic infection and inflammation. Interestingly, four bnabs specific for the HIV- CD4 binding site (CD4bs) (VRC0, VRC02, CH06, and CH03) bind a common human autoantigen, ubiquitin ligase E3A (UBE3A). The UBE3A protein competitively inhibits gp20 binding to VRC0 and, remarkably, avidity for UBE3A among these four bnabs was correlated with neutralization breadth. The absence of poly- or autoreactivity suggests that non-neutralizing HIV- Abs may dominate responses because they not be subject to control by immunological tolerance. The rarity and delayed development of CD4bs bnab in infected individuals may be the consequence of viral mimicry of human UBE3A. NON-ABSTRACT DRIVEN 9

20 Tuesday, 28 October Symposium 02: Advances in Animal Models SY02.0 Non-Human Primate Models for New Prevention Strategies of HIV Transmission Roger Le Grand CEA, IDMIT, Fontenay-aux-Roses, France SY02.03 Prevention Studies in Nonhuman Primate Models Jeffrey Lifson National Institutes of Health, National Cancer Institute, Bethesda, MD, United States NON-ABSTRACT DRIVEN Nonhuman primate (NHP) studies significantly contribute to the understanding of HIV sexual transmission mechanisms and to the development of approaches aimed at preventing infection. In this presentation we will review studies in NHP for assessing microbicides and vaccines efficacy. Future directions for preventive strategies will be discussed. SY02.02 How Mice to Monkeys Inform Human B Cell Repertoire Responses Gunilla B. Karlsson Hedestam Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology, Stockholm, Sweden Continued efforts to evaluate HIV- envelope glycoprotein (Env)-based vaccines in vivo are needed to identify immunogens and immunization regimens that elicit responses that protect against HIV- acquisition. HIV- is a highly neutralization-resistant virus due to conformational and glycan shielding of conserved Ab determinants making the development of an effective vaccine extremely challenging. Understanding current obstacles to elicit broadly neutralizing Abs that target poorly accessible epitope on the native Env spike requires the use of both small animal models and non-human primates (NHPs). Among small animals, mice are most useful for basic mechanistic studies due to the ease by which they can be manipulated, the availability of numerous knockout strains, shorter experimental cycles and lower costs. On the other hand, NHPs are more similar to humans, both in terms of the genetics of antibody gene segments and the characteristics of immune cell subsets. NHPs are also more amenable to detailed studies of vaccine-induced responses due to the larger sample volumes that can be obtained and the possibility to perform SHIV challenge studies, but they are costly. Importantly, the choice of animal model should be determined based on the question to be addressed to ensure maximal information output while yet adhering to the 3R principle. Over the past several years, we have established unique methodology to characterize B cell responses in both mice and non-human primates including the isolation of monoclonal Abs from single-cell sorted memory B cells. These studies confirm that an in-depth understanding of the principles that govern B cell activation, differentiation and affinity maturation is needed to accelerate the development of a vaccine capable of stimulating broadly neutralizing antibody responses against HIV-. Such studies will require several types of animal models including both mice and monkeys. SY02.04 The Role of Humanized Mice in Advancing HIV Vaccine and Eradication Studies Todd M. Allen Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States Efforts to determine how human immune responses to HIV can be induced or augmented would be greatly facilitated by the availability of a suitable small animal model. There have been a number of recent advances in the development of small animal models for HIV, including the recent generation of humanized BLT (bone marrow, liver, thymus) mice where implanted human fetal CD34 + hematopoietic stem cells (HSCs) become educated within transplanted autologous human thymic tissues. This model has dramatically improved the ability of humanized mice to support productive HIV replication and CD4+ T cell depletion, as well as recapitulate human cellular and humoral responses to HIV. Here we discuss recent data illustrating the ability of this model to support HIV-specific and vaccine-induced immunity to HIV. We will also discuss applications of this model for elucidating critical aspects of HIV transmission, dissemination, and HIV-specific immunity as well as the opportunities and limitations of this model to support HIV vaccine and eradication efforts. 20 HIV Research for Prevention 204 HIV R4P

21 Symposium 03: Treatment as Prevention: The Promise and the Perils Tuesday, 28 October SY03.0 No Time to Lose - The Importance of Early HIV Infection Kimberly Powers The University of North Carolina at Chapel Hill, Department of Epidemiology, Chapel Hill, NC, United States Detection of early HIV infection is important for individual and public health. This talk will review opportunities and challenges related to the detection of early infection, as well as the implications of ongoing transmission during this critical window. SY03.02 A Tale of Four Treatment-as-Prevention Trials in Africa Frank Tanser,2, Collins Iwuji,3 University of KwaZulu-Natal, Africa Centre for Health and Population Studies, Mtubatuba, South Africa, 2 University of KwaZulu-Natal, School of Nursing and Public Health, Durban, South Africa, 3 University College London, London, United Kingdom SY03.03 Treatment as Prevention - How Can We Predict Success? Deenan Pillay,2, Till Barnighausen, Tulio De Oliveira, Kobus Herbst, Frank Tanser Africa Centre for Health and Population Studies, University of KwaZulu-Natal, Mtubatuba, South Africa, 2 University College London, Division of Infection and Immunity, London, United Kingdom With major trials now ongoing to explore the impact of a treatment as prevention approach to HIV, it is critical that we consider the factors which will predict and determine success, as defined by a reduction in HIV incidence. We will consider three specific areas. Firstly, a prerequisite for TasP impact is the linkage to and retention in care for those diagnosed with HIV. The ability to deliver such linkage is critical to the success of TasP. Secondly, we should expect an increase in absolute numbers of individuals harbouring drug resistant viruses, commensurate with an increasing coverage of antiretroviral treatment. On the one hand, this may become a major impediment to maintaining a reduction in incidence. By contrast, this could merely be a surrogate of treatment coverage, with the benefits of therapy far outweighing the negative impact of resistance mutants. Thirdly, within the now established era of pathogen genomics, we have the capability to monitor the dynamics of spread of HIV through a sampling frame for virus gene sequences. We speculate that such approaches can predict a reduction of incidence in the context of TasP, although this remains to be formally demonstrated. SY03.04 TasP Promises But What Is The Reality? Valerie C Delpech, Alison E Brown Public Health England, HIV & STI, London, United Kingdom The clinical evidence of TasP is compeling and modelling studies are inspirational but what is the reality? Can TasP reduce population level incidence? In this presentation we assess the impact of antiretroviral treatment on HIV incidence, prevalence and undiagnosed infections using examples from the UK, South Africa and other countries. The importance of good HIV surveillance data including testing data, HIV diagnoses by key groups, link to and retention in care, ART uptake and viral load data are critical in monitoring our successes. NON-ABSTRACT DRIVEN 2

22 Tuesday, 28 October Symposium 04: Building Combination Prevention Trials SY04.0 Building Combination Prevention: Trial Designs Helen Weiss London School of Hygiene & Tropical Medicine, MRC Tropical Epidemiology Group, London, United Kingdom Combination prevention trials are based on synergies of multiple prevention and treatment components, including interventions that aim to change structures and environments. Such interventions are by nature community-based rather than individually-based, and thus lend themselves to community-randomized trials rather than individualrandomized trials.the impact of the intervention is then also assessed at population-level. The gold standard method for such evaluations is the parallel-arm cluster randomised controlled trial (RCT), which is logistically challenging, time-consuming, and expensive to implement. We will discuss challenges of the cluster RCT, including the choice of control arm in light of evolving standards of care for HIV treatment and prevention, minimizing contamination between clusters, and methods of randomisation to minimise imbalance between arms.we will also discuss alternative designs and show examples of trials which have evaluated the impact of community-level interventions including stepped-wedge designs and adaptive designs. Methodological challenges in evaluating combination prevention interventions will also be discussed, including time-series models for surveillance data, hierarchical models to allow for clustering in the design, and the use of causal inference methods which are particularly useful for non-randomized designs SY04.02 Combination Prevention: From Trials to Implementation Sinead Delany-Moretlwe University of the Witwatersrand, Wits Reproductive Health & HIV Institute, Johannesburg, South Africa SY04.03 The Role of Mathematical Modeling to Design, Conduct, Interpret and Replicate HIV Prevention Trials Marie-Claude Boily Imperial College London, Infectious Diseases Epidemiology, London, United Kingdom Current prevention trials often aim to test combinations of different prevention and treatment components introduced simultaneously or sequentially in different risk populations. Choosing the best combination package and deciding in which population it should be tested can be challenging. Designing and interpreting the results of costly and large community-based randomized trials (C-RCT) to evaluate complex HIV prevention strategies can be equally challenging. Mathematical models are widely used to predict the population-level impact of interventions and inform policy decisions. In this talk, we will discuss why and how mathematical models are also increasingly and innovatively being used at the different stages of the clinical trial process: to inform product development, design the intervention package, to help design and conduct clinical trials, and to interpret and help generalize results. In so doing, we will also discuss how mathematical models can be used to address some of the challenges encountered in community-based randomized trials. SY04.04 Scalable Adherence Interventions Terrence F. Blaschke,2 Bill and Melinda Gates Foundation, Discovery and Translational Sciences, Seattle, WA, United States, 2 Stanford University School of Medicine, Internal Medicine, Stanford, CA, United States NON-ABSTRACT DRIVEN Combination HIV prevention is the application of several evidencebased interventions including biomedical, behavioural and structural interventions to maximise population-level reductions in HIV incidence in a specific setting. While strong evidence exists now for several single approaches, evidence for how best to combine these different interventions for maximal impact on HIV in different populations and settings is scarce. The success of combination HIV prevention rests on the identification of populations or places with highest rates of infection, adaptation of interventions that are cost-effective and likely to have highest impact on HIV in that population, and delivery of that combination of interventions with sufficient scale, uptake and sustained use. In this talk, we discuss how using a public health approach can guide the pragmatic, localised application of evidence-based combination approaches for particular populations. We highlight how advances in fields as diverse as HIV phylogenetics and molecular epidemiology, market research and implementation science may refine our understanding of local transmission dynamics and enhance our ability to deliver feasible and acceptable interventions successfully and with maximum impact. In addition to monitoring HIV incidence trends, the importance of monitoring the potential synergistic or antagonistic effects of these interventions in particular populations, and the need for responsive programming in order to minimise unintended consequences, will also be addressed. Recognizing that suboptimal adherence is the major cause of lack of effectiveness of ARVs for PrEP, scalable interventions to improve adherence are needed. Many interventions have been proposed, but to date none have been scalable and sustainable (Medication Adherence Interventions, Evidence Report No AHRQ Publication No.2- E00-EF, 202). A limitation of most studies is the absence of reliable measures of adherence before and after an intervention. Moreover, the duration of benefit in those studies showing some increase in adherence dissipates over several months. What is needed is a scalable approach to identifying suboptimal adherence, monitoring those more likely to continue or become poorly adherent due to known predisposing factors, then focusing interventions on that cohort of patients. Due to the correspondence of drug exposure to protection from infection, sparse sampling of dosing information is insufficient, and detailed dosing information itself, shared with the patient and the provider, can significantly improve adherence. Modern technology allows detailed dosing histories to be obtained unobtrusively and collected centrally at a point in time when interventions can be applied. Combined with approaches such as Managed Problem Solving (Gross et al., JAMA Intern Med. 203; 73: ) and Lifetime HIV Antiretroviral Therapy Adherence Intervention: Timing Is Everything (Bangsberg and Haberer, JAMA Intern Med. 203; 73:306-7), progress towards scaling interventions to large high-risk populations is now within reach. 22 HIV Research for Prevention 204 HIV R4P

23 Wednesday, 29 October Symposium 05: The Role of Structure Based Design in Vaccine Development and Immunoprophylaxis SY05.0 Structure-Based Design of Improved Antibodies for HIV- Treatment or Prevention Louise Scharf, Stuart A. Sievers, Anthony P. West, Jr., Ron Diskin,2, Michel C. Nussenzweig 3, Pamela J. Bjorkman California Institute of Technology, Division of Biology and Biological Engineering, Pasadena, CA, United States, 2 (present address) Weizmann Institute of Science, Department of Structural Biology, Rehovot, Israel, 3 Rockefeller University, Laboratory of Molecular Immunology, New York, NY, United States Over 30 years after the emergence of HIV-, there is no effective vaccine, and AIDS remains an important threat to global public health. Following infection by HIV-, the host immune response is unable to clear the virus due to a variety of factors, including rapid viral mutation and the establishment of latent reservoirs. The only target of neutralizing antibodies is the trimeric envelope spike complex, but HIV- can usually evade anti-spike antibodies due to rapid mutation of its two spike glycoproteins. We are using structure-based protein design methods to engineer antibodies that can resist some of the common routes of HIV- mutation, with the hope that the designed antibodies could be used in passive immunotherapy methods for HIV- treatment and/or prevention. SY05.02 The Stucture of the Glycan Shield of HIV Max Crispin University of Oxford, Biochemistry, Oxford, United Kingdom SY05.03 Structure-Based Stabilization of the Prefusion Closed HIV- Env Trimer Peter Dak Pin Kwong National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States The HIV- envelope spike (Env) assumes diverse conformations to facilitate dual roles in viral entry and immune evasion. An antigenically specific immunogen which only binds to broadly neutralizing antibodies and not to non-neutralizing or poorly neutralizing antibodies might lead to increased elicitation of effective HIV--neutralizing antibodies. Here, we report the mature unliganded crystal structure at 3.7 Å resolution, for one of the most antigenically specific Env immunogens developed to date, the SOSIP.664 variant of the BG505 strain of HIV-. The unliganded structure closely resembled recently determined antibody-bound structures of BG505 SOSIP.664 and appeared to be in the prefusion closed state. Because SOSIP.664 retains the ability to transition to an open CD4-bound state, which is recognized by poorly neutralizing antibodies like the CD4- induced antibody 7b as well as antibodies against the immunodominant V3 region like D, we used structure-based design to identify mutants with reduced binding to 7b and D. Select variants with introduced disulfide bonds or proline mutations no longer bound CD4-induced or V3-directed antibodies, even in the presence of CD4. Unexpectedly, these variants did retain the ability to bind to CD4, with electron microscopy reconstructions indicating them to remain in a closed state, even when bound by CD4. Structure-based design can thus be used to improve the antigenic specificity of HIV- Env, with conformationally fixed variants allowing for detailed mechanistic dissection of ligand-induced transitions. SY05.04 Reductionist Vaccine Design to Induce Broadly Neutralizing Antibodies against HIV William Schief,2,3 Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA, United States, 2IAVI Neutralizing Antibody Center at The Scripps Research Institute, La Jolla, CA, United States, 3Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States One of the major thrusts of our work on HIV vaccine development has been to define immunogens and regimens to induce VRC0-class broadly neutralizing antibodies. In this work we have defined a reductionist approach to vaccine design that could be applied to other epitopes on HIV and other pathogens. We will review this line of research, and weigh opportunities and challenges to induce broadly neutralizing antibodies against the VRC0 epitope and other conserved HIV epitopes, in light of recently published structures of the HIV trimeric spike. NON-ABSTRACT DRIVEN 23

24 Wednesday, 29 October Symposium 06: Harnessing Antibody Effector Functions SY06.0 Evaluating HIV- Vaccine Induced Antibody Effector Function to Advance Vaccine Candidates Georgia Tomaras Duke University, Duke Human Vaccine Institute, Departments of Surgery, Molecular Genetics and Microbiology, and Immunology, Durham, NC, United States SY06.03 Human Non-neutralizing HIV- Envelope Monoclonal Antibodies Limit the Number of Founder Viruses during SHIV Mucosal Infection in Rhesus Macaques Sampa Santra Beth Israel Deaconess Medical Center, Harvard Medical School, Center for Virology and Vaccine Research, Boston, MA, United States This presentation will discuss how to utilize the diverse array of humoral measurements to evaluate and inform decisions for advancing HIV- vaccine candidates. Antigen specific antibody responses, some of which are Fc-mediated, correlate with decreased risk of HIV- infection. Vaccine induced polyclonal antibody responses and antibody effector function differ profoundly across vaccine regimens. Knowledge of protective and potentially detrimental immune responses can be garnered by systematically examining the correlations between such responses and risk of HIV- infection after exposure. Currently, the HIV- vaccine field has an array of humoral immune correlates of risk obtained from both human efficacy studies, as well as from nonhuman primate studies. Analyses that delineate the interplay among immune responses (innate, cellular, and humoral), host genetics (e.g., FcR expression, HLA), and HIV- virus diversity (i.e., within the populations/ geographic regions enrolled in vaccine efficacy studies) are likely to be transformative for the next phase of HIV- vaccine design and evaluation. New data for understanding and evaluating vaccine induced antibody breadth and effector function will be presented. SY06.02 A Systems Serology Approach to Vaccination Margie E. Ackerman, Chris Bailey-Kellogg 2, Galit Alter 3 Dartmouth College, Thayer School of Engineering, Hanover, NH, United States, 2 Dartmouth College, Computer Science, Hanover, NH, United States, 3 Massachusetts General Hospital, Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, United States SY06.04 Combating Cell-Cell Transmission of HIV with Antibodies - Slaying the Trojan Horse? Stephen Kent University of Melbourne, Department of Microbiology and Immunology, Peter Doherty Institute, Melbourne, Australia Developing vaccines with broad neutralizing function is a holy grail of HIV research, but there is a potential problem with this approach. HIV is readily isolated from cells within the semen of HIV-infected men, and this cell-associated virus (dubbed Trojan horse leukocytes ) is more infectious than free virions within semen. Data emerging from clinical studies in humans and studies conducted in the non-human primate model of HIV infection provide evidence for a role of cell-associated virus in HIV transmission. The extent to which neutralizing antibodies or MHC-restricted CTLs can stop cell-cell virus transmission in unclear. ADCC antibodies can however readily recognize foreign infected cells and may therefore assist in preventing cell-cell transmission of HIV. We hypothesize ADCC against infected cells may be one mechanism for the partial efficacy of the RV44 trial. NON-ABSTRACT DRIVEN In vivo, antibody responses to vaccination or natural infection are highly polyclonal, with multiple somatic variants directed to multiple epitopes on multiple antigens. This diversity of variable domain recognition characteristics is further complemented by diversity in the constant domain s ability to interact with innate immune receptors such as FcgR. Antibody-based protection is thus derived from the sum of specificities and activities of this polyclonal humoral milieu. We demonstrate a highthroughput, high-content platform for the biophysical interrogation of the innate immune recruiting capacity of polyclonal, antigen-specific antibodies capable of parsing the complex humoral milieu into components that can be associated with relevant clinical, genetic, or functional characteristics. Thousands of measurements of the humoral immune response can be determined, providing a comprehensive landscape of antibody activity and better understanding of the characteristics and development of both protective and pathological humoral immunity that may be critical to understanding vaccine-mediated protection. When applied to chronically HIV infected or vaccinated subjects, dramatically different antibody profiles are observed, and these profiles can be used to develop computational models of antibody activity that provide insight into mechanisms of vaccination. 24 HIV Research for Prevention 204 HIV R4P

25 Wednesday, 29 October Symposium 07: Novel Formulations and Sustained Delivery of Antiretrovirals SY07.0 Injectable and Implantable Antiretroviral Strategies for HIV Prevention Ian McGowan University of Pittsburgh School of Medicine, Department of Medicine, Pittsburgh, PA, United States SY07.03 Novel On-Demand Vaginal Microbicide Formulations: Tablets and Films Jill Schwartz CONRAD Eastern Virginia Medical School, Arlington, VA, United States A fundamental challenge associated with the use of antiretroviral preexposure prophylaxis (PrEP) is that individuals must be willing to selfadminister such products daily or in a peri-coital fashion for the strategy to be effective. Multiple Phase 2B/3 prevention trials have demonstrated that adherence to these regimens can be sub-optimal and consequently PrEP interventions have had highly variable effectiveness depending on the population studied and their degree of product adherence. In trials where PrEP adherence has been high, the intervention has been highly effective. Long acting injectable or implantable products are currently used for contraception and this strategy is being evaluated for delivery of antiretroviral drugs. This approach has the potential to provide antiretroviral PrEP for up to 3 months with a single injection and to circumvent adherence problems associated with daily or per-coital administration of oral/topical PrEP agents. TMC278 LA (rilpivirine) and GSK 744 (cabotegravir) have both completed Phase evaluation and are moving into later stage development. The purpose of this talk is to review these two products as well as the potential for the development of long acting single antiretroviral and combination antiretroviral/ contraceptive implantable products for HIV prevention. SY07.02 Intravaginal Rings for HIV Prevention in Women Thesla Palanee-Phillips Wits Reproductive Health and HIV Institute, Johannesburg, South Africa Of the more than 35 million people living with HIV, half are women. Most women acquire HIV through heterosexual intercourse. In fact, women are twice as likely as men to acquire HIV during vaginal sex, due in part to biological factors that make them more susceptible. Young women are especially vulnerable. Efforts to promote abstinence, monogamy and the use of male condoms have not been enough to stop the epidemic, nor are these approaches practical in many settings. In southern Africa, young women are up to five times more likely to become infected with HIV than young men. Microbicides are products applied inside the vagina or rectum to protect against HIV though sex. Although microbicides are not yet available for widespread use, researchers are making significant strides in the development and clinical evaluation of both vaginal and rectal microbicide products. Microbicides that incorporate antiretroviral (ARV) drugs are showing particular promise. A vaginal microbicide could potentially give women the means to protect themselves against HIV. Vaginal microbicides are being designed in many forms, including gels, films and rings, which release an active ingredient gradually over time. Two products are currently in Phase III trials. Vaginal rings are products designed to allow for the slow delivery of a drug or multiple drugs to cells inside the vagina over a period of weeks or months. One is testing a vaginal gel containing the ARV tenofovir used before and after sex, while two trials are evaluating a vaginal ring containing dapivirine that women use for a month at a time. I will focus on the use of intravaginal rings in HIV prevention research. SY07.04 Nanotechnology for Drug Delivery Kim A. Woodrow University of Washington, Bioengineering, Seattle, WA, United States The delivery of drug combinations is a paradigm for treatment of HIV/ AIDS, cance and drug resistant bacterial infections. My laboratory is interested in the application of engineered nanomaterials to control the spatial and temporal delivery of a combination of agents (small molecules, biologics, and conjugates). Strategies to combine chemically incompatible agents may facilitate the discovery of unique drug-drug activities, particularly unexplored combination drug synergy. In this presentation, I will summarize our efforts to develop polymeric delivery systems for the combination delivery of antiretroviral (ARV) drugs for HIV prevention and treatment. ARV drug combinations have the potential to enhance the efficacy of current prevention strategies by overcoming low user adherence, and harnessing drug combinations with synergistic activity and breadth of coverage against the global diversity of HIV variants. We have developed polymeric particulate and fiber carrier systems for delivering ARV drug combinations. The flexibility to design the nanoarchitecture of these polymeric carriers, combined with the versatility of drugs that can be encapsulated for controlled release, motivate the use of these systems for topical, injectable or oral delivery of combination agents in the fight against HIV. NON-ABSTRACT DRIVEN 25

26 Wednesday, 29 October Roundtable 0: Safer Sex In 204: Has the Paradigm Shifted? RT0.0 What Is Safer Sex in 204? Understanding the Biology of HIV Prevention James A. McIntyre,2 Anova Health Institute, Johannesburg, South Africa, 2 University of Cape Town, School of Public Health & Family Medicine, Cape Town, South Africa RT0.02 Communicating a New Safer Sex Paradigm: The Science of Communication Lebogang Ramafoko Soul City, South Africa RT0.03 Missing the Future is not an Option Jim Pickett AIDS Foundation of Chicago, Prevention Advocacy/IRMA, Chicago, IL, United States It s not an opinion, it s a fact. The paradigm of what safer sex means has changed. Because we now have successful, scientifically proven biomedical prevention strategies, safer sex = condoms has evolved to safer sex with a drop-down menu. In the United States, not everyone is comfortable with this expansion of prevention choices. Speaking to an assembly in Frankfurt, Germany in 963, John F. Kennedy said, Change is the law of life. And those who look only to the past or the present are certain to miss the future. So yes, change is the law. It s good that the law is also to buckle up, because the evolution of safer sex has been a bumpy ride so far, and continues to be. What is this all about? Can we ride it out? What needs to be done to smooth our way now and better prepare for the new biomedical tools we will certainly have? Staying stuck in the past and mired in the present isn t going to get us anywhere close to zero. RT0.04 Policy and Program: Perspectives on Safer Sex Nduku Kilonzo, LUCT, Kenya NON-ABSTRACT DRIVEN 26 HIV Research for Prevention 204 HIV R4P

27 Wednesday, 29 October Roundtable 02: Prevention of Mother-to-Child Transmission Revisited RT02.0 The Role of PMTCT and Very Early Treatment in the Elimination of Pediatric HIV Infection Sarah W Read NIAID, Division of AIDS, Rockville, MD, United States RT02.03 Prevention of Mother-to-Child Transmission Revisited Louise Kuhn Columbia University, New York, NY, United States RT02.02 The Case for Early Antiretroviral Treatment towards HIV Cure in Perinatal Infection Deborah Persaud Johns Hopkins University School of Medicine, Baltimore, MD, United States The reservoir for HIV in resting memory CD4+ T cells precludes cure with current antiretroviral drugs. Substantially limiting the size of the latent reservoir is the first step towards drug free remission and cure. In perinatal HIV infection, early antiretroviral treatment significantly alters the size of latent viral reservoirs. The unique aspects of the neonatal immune system coupled with the potential to initiate very early treatment therefore provide a unique opportunity to assess very early treatment and drug-free remission in neonates. This is exemplified in the case of the Mississippi Child who experienced 28 months of drug free remission following 8-months of antiretroviral treatment starting at 30 hours of life. The rationale for clinical trials aimed at achieving drug free remission and cure in perinatal HIV infection will be discussed. In light of recent data on the challenges of curing HIV infected babies, this session will review current approaches in PMTCT and how these may need to be modified. Conceptual distinctions between prevention and treatment in the maternal-infant context will be made and the special issues related to each discussed. RT02.04 PMTCT: Mind the Gaps! Lee Fairlie Paediatrics, Wits RHI, Johannesburg, South Africa Although huge gains have been made in PMTCT with significant reductions in the numbers of HIV-infected infants, there are a few areas that still need attention to improve care for women and their infants to ensure that current gains continue. This discussion will include where the current programmatic and research gaps are and highlight where there may be failures, particularly related to high-risk populations. NON-ABSTRACT DRIVEN 27

28 Thursday, 30 October Roundtable 03: Diffusion of Innovation: Accelerating Along the Research to Rollout Continuum RT03.0 A Case Study of Circumcision Agnes Binagwaho Ministry of Health, Rwanda RT03.04 Diffusion of Innovation: A South African Perspective Yogan Pillay Department of Health, South Africa RT03.02 Male Circumcision and Pre-Exposure Prophylaxis (PreP) Roll Out in Kenya: A Tale of Two Innovations Peter Cherutich Ministry of Health, Nairobi, Kenya RT03.05 From Research Result to Public Health Impact: What Have we Learned and How Can We Do it Better and Faster Mitchell J. Warren AVAC, New York, NY, United States RT03.03 HIV Prevention: Moving from Evidence to Demonstration Projects to Policy and Programs The presentation will focus on historical experiences along the researchto-rollout continuum in HIV prevention and related fields to outline specific lessons that could be applied going forward with upcoming HIV prevention research results. These lessons will be used to explore what should be done in future product introduction, who should be doing it, and when different aspects of rollout should take place. Connie Celum University of Washington, Department of Global Health & Medicine, Seattle, WA, United States NON-ABSTRACT DRIVEN Recent oral PrEP trials demonstrate that biomedical prevention interventions have strong behavioral components in terms of uptake and adherence. It is important to understand user perspectives about antiretroviral-based prevention which impact product use; relevant disciplines for this research are user-centered design and mental models. When products are demonstrated efficacious, diffusion of innovation research and social marketing can be used to design demand creation and delivery models. Provider perspectives need to be understood and factored in designing models for delivery of new technologies, training needs, and policies. There are important opportunities to learn from implementation of novel HIV prevention technologies as they are shown to be efficacious. Demonstration projects are important to evaluate communication strategies about efficacy and decision tools to help potential users decide about product use, acceptable and feasible strategies to support adherence, and cost-effectiveness of delivery. These demonstration projects should be conducted in parallel with clinical development of sustained release and less user-dependent formulations, so that a mix of HIV prevention methods can be offered to persons at risk. 28 HIV Research for Prevention 204 HIV R4P

29 Roundtable 04: Research Where and With Whom it Matters Thursday, 30 October RT04.0 MSM: Long Road to Trial Participation in Africa Eduard J. Sanders,2,3 KEMRI-Wellcome Trust Research Programme, Kilifi, Kenya, 2 University of Oxford, Headington, United Kingdom, 3 Amsterdam Institute for Global Health and Development, Amsterdam, Netherlands RT04.03 HIV Prevention Trials in Developing Countries: Challenges and Opportunities Fareed Abdullah South African National AIDS Council, South Africa RT04.02 We are not Guinea Pigs: HIV Clinical Research for Smarties Udom Likhitwonnawut AVAC, Chiang Mai, Thailand RT04.04 Engaging Adolescents and Young Adults in HIV Prevention Trials Sybil Hosek Stroger Hospital of Cook County, Psychiatry, Chicago, IL, United States HIV-related stigma makes HIV research on marginalized populations in Thailand challenging, particularly in studies involving populations with a long-standing history of persecution and ill-treatment such as drug users and sex workers. Despite the low HIV prevalence in the general Thai population, HIV prevalence and incidence among injecting drug users and MSM are consistently higher than other groups. For this reason, these groups have become favorite subjects of various HIV prevention studies. Involving criminalized, persecuted populations in clinical trials can lead to disputes and opposition from various stakeholders. In Thailand, there are some studies involving these marginalized populations that did not invest sufficient resources and time working with other stakeholders. As the result, the studies encountered opposition from stakeholders. The opposition often led to spending more time and resources to enroll trial participants, discrediting the results of the study, and a feeling of distrust between the stakeholders. Respect and adherence to ethical research principles can help researchers to overcome the resistance and disputes caused by the study. How and where a study is conducted and staffed can significantly affect informed consent process and the ability of potential trial participants to make free decisions. The principle of beneficence and do no harm can guide researchers on how to provide the best HIV prevention methods to trial participants. Applying other HIV ethical guidelines such as Good Participatory Practice (GPP) can also strengthen community engagement by involving community members in HIV trials and help create meaningful community participation in HIV research, as well as reducing stigma, and increasing knowledge and understanding about HIV clinical research. Lastly trial sponsors and funders have a responsibility in site selection to ensure that the sites have the capacity and capability for meaningful stakeholder engagement. RT04.05 African Couples Testing Matters: Most Transmissions Are in Marriage, Most Pregnancies Are Not Immaculate Conceptions, and Women Aren t Just Incubators Susan Allen, William Kilembe, Mubiana Inambao, Amanda Tichacek, Eric Hunter 2, Rwanda Zambia HIV Research Group Rwanda Zambia HIV Research Group-Emory University, Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA, United States, 2 Emory University, Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA, United States More than 20 million Africans have been tested for HIV. Despite clear evidence that the majority of transmissions in Africa occur within stable heterosexual couples often during pregnancy from husbands not known to be HIV positive fewer than 5% of couples have been tested and counseled together. In this context, and because two people are required for HIV transmission, we argue that all prevention efforts and therapy should start with Couples Voluntary HIV Counseling and Testing (CVCT). CVCT is effective and affordable. CVCT is endorsed by the WHO and training materials are available from the CDC in several languages. The US government has recently funded large-scale trials of treatmentas-prevention (TasP), yet compared to CVCT which prevents one HIV infection for less than $500, TasP would cost $7000 per year. Most African countries have per capita incomes of less than $500 and annual per capita health expenditures less than $00. In this setting, CVCT should be a priority to achieve cost effective HIV prevention. Moreover, in spite of CVCT effectiveness and affordability, some population TasP trials deliberately withhold CVCT from African couples. This behavior is of questionable ethics. CVCT should be provided to all couples as standard of care. The impact and cost-effectiveness of CVCT and TasP should be assessed separately. NON-ABSTRACT DRIVEN 29

30 Thursday, 30 October Symposium 08: Mucosal Barriers to Infection SY08.0 Upper Female Genital Tract and Defining Safety Ruth M. Greenblatt University of California San Francisco, Clinical Pharmacy, Medicine, Epidemiology and Biostatistics, San Francisco, CA, United States SY08.03 Mechanisms of HIV- Restriction and Dissemination Mediated by Intestinal Mucosal Myeloid Cells Gabriella Scarlatti, Mariangela Cavarelli San Raffaele Scientific Institute, Milan, Italy Multiple tissue sites of susceptibility to HIV infection are present in the female genital tract. The specific portal of entry of virus may depend on a range of circumstances including concurrent STIs, trauma, hormonal exposures, male factors and the influence of topically applied substances. The upper FGT, including the endocervix and endometrium, are potential sites of vulnerability and should be considered in the evaluation of prevention modalities. Additionally, many products, including commonly used topical vehicles and hygiene aids, underwent regulatory evaluation prior to the current era of sophisticated mucosal immunologic research. These products and substances may need reappraisal using modern methods, since adverse effects may occur. SY08.02 Behavioral and Biological Factors Affecting HIV Acquisition in Women Douglas Kwon Ragon Institute of MGH, MIT and Harvard, Massachusetts General Hospital and Harvard Medical School, Cambridge, MA, United States The infection of specific cellular niches by HIV- adopting mechanisms of restriction of replication are a challenge to develop effective prophylactic and therapeutic interventions. Specifically, the mucosal surfaces are the predominant sites involved in the first steps of viral uptake and entry, and CD4+ T cell depletion. There is increasing evidence that dendritic cells (DC) and macrophages residing in the lamina propria of the mucosa may be the first cellular targets mediating transmission. We have recently provided the proof of principle of the active involvement of intestinal lamina propria resident CDc+ DCs in the uptake of HIV. Indeed, gut DCs migrate towards and extend processes between the tight junctions of columnar epithelium through a CCR5-dependent mechanism, take up R5 but not X4 virus, and then transfer infection to T cells. These data raise the question on which DCs subsets are mediating infection and, in turn may affect the fate of the virus and the immuneresponse. SY08.04 Rectal Microbicide Development: An Update Ross D. Cranston University of Pittsburgh, Pittsburgh, PA, United States NON-ABSTRACT DRIVEN 30 HIV Research for Prevention 204 HIV R4P

31 Symposium 09: Sustaining Durability of Responses Thursday, 30 October SY09.0 Clinical Translation of HIV Protein-Expressing Cytomegalovirus Vectors Louis Picker Oregon Health & Science University, Vaccine and Gene Therapy Institute, Beaverton, OR, United States SY09.03 Adjuvants and Durability of Vaccine-Induced Immunity Marguerite Koutsoukos GlaxoSmithKline, Vaccines Discovery & Development, Rixensart, Belgium SY09.02 Antibody Persistence and T Cell Balance: Two Key Factors Confronting HIV Vaccine Development George K. Lewis, Tony L. Devico, Robert C. Gallo Institute of Human Virology, University of Maryland School of Medicine, Division of Basic Science and Vaccine Research, Baltimore, MD, United States The quest for a prophylactic AIDS vaccine is ongoing but it is now clear that the successful vaccine must elicit protective antibody responses. Accordingly, intense efforts are underway to identify immunogens that elicit these responses. Regardless of the mechanism of antibodymediated protection, be it neutralization, Fc-mediated effector function, or both, antibody persistence and appropriate T cell help are significant problems confronting the development of a successful AIDS vaccine. Evidence will be presented illustrating the poor persistence of antibody responses to Env, the envelope glycoprotein of HIV-, and the related problem of CD4+ T cell responses that compromise vaccine efficacy by creating excess cellular targets of HIV- infection. Finally, solutions to both problems will be proposed that are applicable to all Env-based AIDS vaccines regardless of the mechanism of antibody-mediated protection. SY09.04 Integrase Defective Lentiviral Vectors for Induction of Persistent and Functional Immune Responses Andrea Cara Istituto Superiore di Sanità, Rome, Italy The development of an HIV- vaccine that elicits durable and broadlyreactive functional antibodies remains challenging but the goal of a vaccine strategy. Integrase-defective lentiviral vectors (IDLV) represent a new and promising delivery system for immunization purposes, endowed with peculiar characteristics, setting them apart from the parental integration-competent lentiviral vectors. Current data suggest that IDLV are able to induce long-lasting and protective immune responses in mice after a single immunization. We recently started a study with the aim to demonstrate in monkeys that immunization with IDLV delivering HIV Envelope protein induces sustained and functional anti-env antibodies (Abs) and T cell responses. The status of this work will be discussed along with new immunization strategies that derive from this work. NON-ABSTRACT DRIVEN 3

32 Friday, 3 October Roundtable 05: Multipurpose Technologies RT05.0 MPTs: What Are they, and Why Are they Needed? Judy M. Manning, Cara J. Chrisman US Agency for International Development, Office of Population & Reproductive Health, Washington, DC, United States RT05.03 Regulatory Considerations with Developing Combination Products Charu Mullick Food and Drug Administration, Silver Spring, MD, United States One of the fastest growing fields in women s sexual and reproductive health (SRH) is multipurpose prevention technologies (MPTs) that simultaneously prevent unintended pregnancy, HIV, and other sexually transmitted infections (STIs). MPTs have the potential to: ) meet women s multiple SRH prevention needs in a single product; 2) achieve efficiencies in cost of delivery of prevention products and services; and 3) leverage existing delivery channels to achieve higher levels of SRH prevention product uptake and demand. This presentation will provide an overview of the public health rationale for MPTs, and the creation of the MPT Initiative, summarize recent efforts to harmonize and prioritize the product development pipeline, and describe high priority MPT products currently in late stage development. RT05.04 Study Designs for Evaluating the Effectiveness of Multipurpose Technologies Elizabeth R Brown,2 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 University of Washington, Biostatistics, Seattle, WA, United States RT05.02 State of the Art: Multipurpose Prevention Technology Science and Product Development Joseph Romano NWJ Group, LLC, Wayne, PA, United States RT05.05 Multipurpose Prevention Technology Elizabeth Bukusi KEMRI, Nairobi, Kenya NON-ABSTRACT DRIVEN The development of multipurpose prevention technologies (MPT) products involves complex and diverse scientific disciplines. Product design and development, clinical evaluation and regulatory approval all present challenges to the field and addressing these challenges will require diverse expertise and innovation. Although there is active development of early stage MPT products, there are also efforts to develop next generation technologies that can address the technical and market-based challenges confronted by current MPT products. This presentation will discuss the status of current MPT development efforts, the development and regulatory challenges these products face, and the ongoing research and development that will improve and enable future MPT products. Multipurpose Prevention Technologies (MPTs) are products under development that are capable of simultaneously addressing women s sexual and reproductive health (SRH) including unwanted pregnancies, STDs such as HIV and other reproductive tract infections. Women s SRH needs are interrelated. Unplanned pregnancies account for nearly half of all pregnancies worldwide and lead to almost 00,000 maternal deaths per year as a result of unsafe abortions and complications of pregnancy and delivery. Women have inadequate access to efficacious contraception especially in areas with high prevalence of HIV. Transmission of HIV and other STIs such as HSV-2 and HPV are highly prevalent among women, especially in some specific locations and in particular pockets of the developing world. The available MPTs, male and female condoms, may not be a realistic option for many women: condom use is inconsistent and in addition issues of power and trust make it difficult for women to negotiate its use. The female condom has not been widely accessible nor affordable. Safe, acceptable and affordable technologies that address these simultaneous RH needs in a more holistic way are urgently needed. MPTs address these shortcomings and build on years of contraceptive and HIV research to develop more comprehensive and effective prevention methods. MPTs additionally address several health priorities identified in the MDGs including: improving maternal health (MDG 5) through access to technologies and interventions that meet the complex range of women s reproductive health needs; and combating HIV/AIDS and other diseases (MDG 6) through technologies that target STIs and inadvertently improving child health (MDG 4) by improving their mothers health. Potential MPTs, both coitally-dependent and long acting are under development. The presentation will update participants on the status of current and future research of technologies and products that are focused on reproductive needs of women 32 HIV Research for Prevention 204 HIV R4P

33 Symposium 0: Challenges of Biomedical HIV Prevention Trials Friday, 3 October SY0.0 HIV Prevention Trials: Their Successes and Failures Jeanne Marrazzo University of Washington, Seattle, WA, United States In the last several years, the HIV prevention field has experienced unprecedented success with new approaches to biomedical prevention, most notably with pre-exposure prophylaxis (PrEP) and medical male circumcision. Tenofovir and emtricitabine, logical candidates for PrEP because of their potent inhibition of the HIV- reverse transcriptase, generally good safety and tolerability profiles in HIV-infected persons, high barrier to HIV- resistance, and favorable pharmacokinetics, have been approved in combination for PrEP and were recently recommended by the WHO for persons at risk for HIV infection, especially men who have sex with men (MSM). Despite these advances, however, the prevention field has been buffeted by several disappointing findings and realizations, mainly the complex interactions between adherence to study product, perception of HIV risk, the social stigma attached to HIV and antiretroviral medication, and the social fabric in which clinical trials are conducted. Both VOICE and Fem PrEP trials successfully enrolled young African women at high risk for HIV, yet adherence was low in both. Moreover, in VOICE, participants with the highest risk profiles were the least likely to adhere to the study drugs. Thus, more accurate adherence measures are critical to estimate healthy participants product use during HIV- prevention trials. Products that require minimal daily adherence, including sustained antiretroviral delivery from vaginal rings or injections, may be more suitable for some women. Alternatively, successful adoption of daily PrEP may require different social and operational strategies to support daily use. Indeed, the high rates of retention we observed suggest that participants valued the study and its contribution to their health. Understanding perceived motivation for and value of study participation and investigational products, and leveraging this when conducting biomedical HIV- prevention research, is critical. SY0.02 Modifiers of PrEP Efficacy: Co-Infections, Sex, Hormones and Microbiome Betsy Herold Albert Einstein College of Medicine, Bronx, NY, United States SY0.03 Enrolling Adolescents in HIV Vaccine Trials: Will We Be Ready? Ann E. Strode, Catherine M Slack 2 University of KwaZulu-Natal, School of Law and HIV/AIDS Vaccines Ethics Group, Pietermaritzburg, South Africa, 2 School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Applied Health Sciences, HIV/AIDS Vaccines Ethics Group, Pietermaritzburg, South Africa Adolescents in sub-saharan Africa are at high risk of HIV infection and the testing of new HIV prevention technologies, such as candidate HIV vaccines, are urgently required as part of a combination prevention approach for this sub-group. Should an experimental HIV vaccine show sufficient efficacy in adults, it is critical that stakeholders are fully prepared for the enrollment of adolescents in efficacy studies. Much has been written on the public health need to enroll adolescents into HIV prevention studies to obtain relevant data that could be used to license new prevention products specifically for this group. Less work has been done to explore children s rights to benefit from scientific progress and whether this includes a right, if any, to participate in such studies. This paper assumes it is both ethically and legally justifiable, in certain circumstances, to include adolescents in HIV prevention trials. However, their enrollment into such trials will be complex because consent, privacy, illegal sexual activity and mandatory reporting obligations (among others) will need to be addressed. This paper, drawing on work funded in part by the NIH (award number RO A094586) sets out key norms for reflecting on these complexities. It asks how adequately prepared countries will be to host adolescent trials, reviews progress towards adolescent enrollment, and discusses a framework that could be used to assess preparedness in South Africa, and even other Southern African countries. It concludes that children, like adults, have a right to benefit from scientific progress. Their age should not be a reason to undermine their rights to access new HIV prevention technologies. However, to fully realise such rights, concerted and sustained attention must be given to promoting their welfare as trial participants. SY0.04 Why Context Matters in Understanding the Challenges to Clinical Trials Jonathan Stadler Wits Reproductive Health & HIV Institute, Johannesburg, South Africa Recently, the uneven results of microbicide trials have drawn attention to the political economy and social and cultural context in which trials are conducted and the significance this has for enrollment, retention, and adherence to study products. Drawing on data from two microbicide trials, my talk presents an anthropological analysis of two recent microbicide / PrEP trials and argues that a holistic perspective of the trial as a social phenomenon may enhance our understanding of the challenges to biomedical trials. The presentation also suggests ways of incorporating social, scientific and ethnographic methods into clinical trials in the endeavor to promote nuanced and thoughtful analysis. NON-ABSTRACT DRIVEN 33

34 Friday, 3 October Symposium : Emerging Areas in Immunity SY.0 Systems Vaccinology: Enabling Rational Vaccine Design with Systems Biology Bali Pulendran Emory Vaccine Center, Atlanta, GA, United States Despite their great success, we understand little about how effective vaccines stimulate protective immune responses. Two recent developments promise to yield such understanding: the appreciation of the crucial role of the innate immune system in sensing microorganisms and tuning immune responses, and advances in systems biology. In this presentation, I will discuss how these developments are yielding insights into the mechanism of some of the most successful vaccines ever developed. Furthermore, such developments promise to address a major challenge in vaccinology: that the efficacy of a vaccine can only be ascertained retrospectively, upon infection. The identification of molecular signatures induced rapidly after vaccination, which correlate with and predict the later development of protective immune responses, would represent a strategy to prospectively determine vaccine efficacy. Such a strategy would be particularly useful when evaluating the efficacy or immunogenicity of untested vaccines, or in identifying individuals with sub-optimal responses amongst high risk populations, such as infants or the elderly. We have used a systems biology approach to identify early gene signatures that correlate with, and predict the later immune responses in humans vaccinated with the live attenuated yellow fever vaccine YF-7D, the seasonal and pandemic influenza vaccines, meningococcal vaccines, pneumococcal vaccines, shingles vaccines and the RTS,S malaria vaccine. I will review these studies, and discuss their broader implications for vaccinology, and highlight some of the critical insights that are emerging. SY.02 Resident Memory CD8 T Cells: Quantity, Location, and Function Kathryn Fraser, Jason Schenkel, Ashley Haase, David Masopust University of Minnesota, Minneapolis, MN, United States SY.03 Universal, MHC-E Restricted CD8+ T Cells Participate in RhCMV Vaccine Vector-Induced Protection Against SIV Helen Wu, Scott G. Hansen, Katherine B. Hammond, Collette M. Hughes, Jason Reed, Abigail B. Ventura, Reesab Pathak, Alfred W. Legasse 2, Benjamin J. Burwitz, Michael K. Axthelm, Klaus Früh, Louis J. Picker,2, Jonah B. Sacha,2 Oregon Health & Science University, Vaccine & Gene Therapy Institute, Beaverton, OR, United States, 2 Oregon Health & Science University, Oregon National Primate Research Center, Beaverton, OR, United States Background: Vaccination with rhesus cytomegalovirus (RhCMV) expressing SIV proteins results in unparalleled protection from pathogenic SIV replication. The RhCMV-induced, SIV-specific CD8 T cells are characterized by wide breadth, high promiscuity (responses present in all vaccinees), and non-canonical epitope targeting. We hypothesized that the breadth and promiscuity of the RhCMV-induced CD8 T cell response result from unique patterns of Major Histocompatibility Complex (MHC) restriction. Methods: RhCMV/gag-vaccinated rhesus macaques (RMs) were MHCtyped by deep sequencing and single MHC-I transferents produced by transfection of the allele of interest into MHC-I-null cell lines. MHC-I transferents were pulsed with a single 5-mer peptide and used as antigen-presenting cells in ICS with PBMC. T cell recognition of SIVinfected targets was assessed by ICS. Results: RhCMV-induced, MHC-I-restricted CD8 T cell responses are restricted by non-classical MHC-E molecules, including universal supertope responses elicited in every vaccinee. MHC-E-restricted CD8 T cells are able to recognize both autologous and heterologous SIVinfected CD4 T cells in vitro. MHC-E is upregulated on the surface of SIV-infected CD4 T cells, in contrast to decreased levels of classical MHC-I molecules. Conclusions: RhCMV elicits high frequency MHC-E-restricted responses that explain the wide breadth, promiscuity, and novel epitope targeting of the induced CD8 T cell response. As these MHC-E-restricted CD8 T cells can recognize naturally processed antigen on SIV-infected cells, they may confer protection from SIV replication that is observed in ~50% of RhCMV-vaccinated RMs. The MHC-E-restricted CD8 T cell response may be particularly effective due to increased levels of MHC-E on the surface of infected cells. RhCMV-induced, MHC-E-restricted CD8 T cell responses represent a novel immune response against retroviruses. NON-ABSTRACT DRIVEN Memory CD8 T cell quantity, location, and function relate to protective efficacy. This presentation will summarize two series of investigations, ) the characterization of resident memory CD8 T cell distribution, differentiation, and function in mice, and 2) the use of heterologous prime-boost-boost vaccination strategies to establish preternaturally robust, polyfunctional, and broadly distributed SIV-specific memory CD8 T cells in rhesus macaques. New functions for resident mucosal memory CD8 T cells will be described; as local sensors of previously encountered antigens that broadcast innate-like alarm signals that activate local humoral, cell-mediated, and innate immunity and confer an antiviral state that can confer near sterilizing immunity against a viral challenge within the mouse female reproductive tract. Additional data will be presented that support the hypothesis that establishing sufficiently abundant and functional memory CD8 T cells within barrier tissues may achieve rapid protective responses at the site of pathogen exposure in the rhesus macaque model of stringent vaginal SIV challenge. 34 HIV Research for Prevention 204 HIV R4P

35 Symposium : Emerging Areas in Immunity Friday, 3 October SY.04 Cooperativity of HIV-specific Cytolytic CD4 T-cells and CD8 T-cells in Control of HIV Viremia Susan Johnson,2, Michael A. Eller,2, Bruce T. Schultz,2, Richard Lu,2, Alexander F. Oster,2, Damien Z. Soghoian 3, Agnes-Laurence Chenine,2, Galit Alter 3, Ulf Dittmer 4, Mary A. Marovich,2, Merlin Robb,2, Nelson L. Michael,2, Diane L. Bolton,2, Hendrik Streeck,2 US Military HIV Research Program, Silver Spring, MD, United States, 2 Henry M. Jackson Foundation for the Advancement of Military Medicine, Silver Spring, MD, United States, 3 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States, 4 Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany Background: HIV-specific CD4 and CD8 T cell responses have been shown to be critical in the control of HIV replication, though it is unknown whether HIV-specific CD4 and CD8 T cells can act in concert. We therefore defined the functional and transcriptional profile of HIVspecific cytolytic CD4 T cells and addressed the cooperativity between both cellular lineages. Methods: We used multicolor flow cytometry and Biomark Fluidigm technology to compare transcriptional, phenotypic and functional profiles of HIV-specific cytolytic CD4, CD4 Th and CD8 T cells. Additionally, we assessed the ability of these cells to inhibit viral replication alone or together with HIV-specific CD8 T cells and how this interaction is complicated by HIV infection. Results: HIV-specific CD4 cytolytic T cells were characterized by a distinct functional and phenotypic profile in comparison to HIV-specific Th CD4 T cells. The expression of CD57, KLRG-, GrzB and Perforin was significantly (p< 0.05) higher on HIV-specific cytolytic CD4 T cells than Th cells, with T-box transcription factors eomesodermin and tbet expressed on HIV-specific cytolytic CD4 T cells. Overall HIV-specific cytolytic CD4 T cells showed transcriptional and phenotypic similarities to HIV-specific CD8 T cells. Interestingly, cytolytic CD4 T cells were able to inhibit HIV replication by an average of 40%, compared to an average of 60% for CD8 T cells at day 3, but became infected by day 7. CD4 and CD8 T cells were also able to control viral replication synergistically, suggesting a model where cytolytic CD4 T cells are involved in the containment of viral replication, but dependent on the overall control of HIV replication by HIV-specific CD8 T cells. Conclusions: Our data suggests that HIV-specific CD4 cytolytic T cells play a role in HIV infection, acting in concert with CD8 T cells to aid in viral control. Overall, our results provide new insights on CD4 cytolytic T cells functional properties and their involvement in HIV control, valuable for vaccine development. NON-ABSTRACT DRIVEN 35

36 Friday, 3 October Symposium 2: Differential Use of Antibodies in Prevention SY2.0 Update on Clinical Development of VRC0 and Second Generation Neutralizing CD4 Binding Site-Specific Monoclonal Antibodies Barney S. Graham Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States SY2.03 HIV Cell-To-Cell Spread and Host Countermeasures Olivier Schwartz Institut Pasteur, Paris, France VRC0 is an HIV- CD4 binding site-specific monoclonal antibody (mab) with potent and broad neutralizing activity. Phase I investigation was initiated in uninfected persons for potential preventive (VRC 602) uses and in HIV-infected subjects for potential therapeutic (VRC 60) uses. The studies evaluate route and dose and the objectives include safety, tolerability and pharmacokinetics (PK). VRC0 is administered on day 0 and at week 4 intravenously (IV) over a dose range of to 40 mg/kg or subcutaneously (SC) at 5 mg/kg. As of 7/5/4, 40 subjects have received 74 doses. VRC0 was well tolerated by both routes with no serious adverse events or doselimiting toxicity. At 20 mg/kg (n=5) or 40 mg/kg (n=4), the mean (± SD) maximum serum concentrations were, respectively, 870±97 and 6±258 mg/l after the first IV dose. In healthy adults at 20 mg/kg (n=3) and 40 mg/kg (n=4), mean 28 day trough serum concentrations were approximately 32.7±9. and 60.6±20. mg/l, after the first IV dose and 40.6±0.7 and 02.6±48.8 mg/l after the second, respectively. The clearance in healthy adults is estimated to be 466±8 ml per day after IV administration over the 5 to 40 mg/kg (n=) dose range. The elimination half-life was similar in healthy and HIV-infected adults with an overall mean value of 4.6±4.8 days. Future development goals include additional studies of VRC0 evaluating dose, route and schedule, and testing efficacy to prevent infection in high-risk infants and adults. The therapeutic potential of VRC0 to diminish viremia during acute infection, prevent viral rebound after treatment interruption, and reduce the viral reservoir during chronic infection will also be evaluated. Second generation CD4 binding sitespecific mabs with higher potency and breadth and extended half-life are being explored for use as single agents or in combination with other mabs and treatment options. SY2.02 Engineering Bispecific Antibodies for HIV Prevention NON-ABSTRACT DRIVEN David Ho, Neal Padte Aaron Diamond AIDS Research Center, New York, NY, United States 36 HIV Research for Prevention 204 HIV R4P

37 Oral Abstract Session 0: B Cell Immunogen Design Tuesday, 28 October OA0.0 Structure-based Design of Trimeric VV2 Antigens Jason Gorman, Yongping Yang, Aliaksandr Druz, Ulrich Baxa 2, Peter D. Kwong NIH/NIAID/VRC, Structural Biology Section, Bethesda, MD, United States, 2 NIH/NCI, Cancer Research Technology Program, Frederick, MD, United States Background: Broadly neutralizing antibodies (bnabs) targeted against VV2 of HIV- Env are among the most prevalent elicited by natural infection and require less somatic mutation than those targeting other sites, making antibodies of this class attractive targets of elicitation for an HIV- vaccine. The quaternary specificity exhibited by VV2 bnabs necessitates the development immunogens that effectively mimic the trimeric cap of the functional native spike. Methods: Recent structural work has defined the trimeric orientation of the VV2 domains at the apex of the viral spike, the VV2 cap. This VV2 trimeric supersite was transplanted onto heterologous trimeric scaffolds and screened for binding to quaternary specific antibodies. Results: Here we present a trimeric VV2 cap which recapitulates the quaternary alignment of the three VV2 domains present in the native spike, as assessed by binding to quaternary specific VV2- directed bnabs. High throughput screening of over a hundred constructs revealed a single trimerization domain capable of presenting an appropriate mimic of the VV2 cap. Antibody binding was observed to multiple strains transplanted onto the trimeric scaffold while gp20s of the same strain did not bind. The transplanted VV2 construct was able to bind VV2-bNAbs from multiple donors and importantly this trimeric construct bound only to a single fab as observed with VV2-directed antibodies on the HIV- Spike. Conclusions: Recent structural studies of HIV- trimers have provided data to allow rational design of quaternary supersite transplants onto scaffolds. High throughput screening with VV2-directed antibodies identified a construct which presents the VV2 cap in an antigenically similar manner to that observed with the BG505.SOSIP. The scaffolded protein provides a simplified, well behaved alternative for use as a probe and a candidate immunogen to focus the immune response on this specific epitope. OA0.02 A Native Linked Soluble Trimer of the HIV- Spike Displaying Antigenic and Structural Mimetic Properties Shailendra K. Sharma, Natalia de Val 2, Shridhar Bale 3, Javier Guenaga, Andrew B. Ward 2, Richard T. Wyatt IAVI Neutralizing Antibody Center at The Scripps Research Institute, Department of Immunology and Microbial Science, La Jolla, CA, United States, 2 The Scripps Research Institute, Department of Integrative Structural and Computational Biology, La Jolla, CA, United States, 3 The Scripps Research Institute, Department of Immunology and Microbial Science, La Jolla, CA, United States Background: Soluble mimetics of the HIV Env spike are of interest for structure/function studies and immunogenic analysis. The BG505 SOSIP trimers are the new structural gold standard, displaying an excellent antigenic profile, molecular homogeneity and stability, allowing 5-6 Å structural resolution. These trimers require furin co-expression, contain unnatural cysteines and are purified by 2G2-affinity chromatography. Many uncleaved gp40 trimers that maintain gp20-gp4 covalent linkage by mutation of the cleavage site are suboptimal in their structural organization, resulting in recognition by non-neutralizing mabs. Methods: Here, we describe covalent peptide linkage of gp20 to gp4 to create uncleaved but well-ordered trimers. The design rationale was to provide flexibility at the cleavage site to allow native rearrangement of gp20 and gp4 trimeric subunits mimicking the endogenous furin cleavage event. We began this process by using a flexible linker using one to three repeats of G 4 S linking JRFL gp20 to gp4. Env deletion at residue 664 increased expression and an E68K mutation was added to restore PG9/6 recognition. The native flexiblelinked (NFL) trimers were expressed in 293F cells and screened by a panel of trimer-preferring bnabs. Results: Little trimer-associated recognition was observed, but introduction of a I-to-P change in gp4 resulted in increased recognition by trimer-preferring bnabs, especially with two repeats of the flexible linker. These trimers were purified by lectin-affinity and size-exclusion chromatography, followed by negative selection, to isolate well-ordered JRFL NFL trimers as confirmed by negative stain EM. Octet binding analysis revealed recognition by most bnabs but low recognition by non-broad mabs that was confirmed by EM 3D reconstructions of very well-ordered trimers. Conclusions: Uncleaved JRFL gp40-nfl2p trimers mimic cleaved trimers in their antigenic properties and represent an exciting new soluble spike design potentially applicable to other Envs. ORAL ABSTRACT SESSIONS 37

38 Oral Abstract Sessions Oral Abstract Session 0: B Cell Immunogen Design ORAL ABSTRACT SESSIONS OA0.03 A Recombinant HIV Envelope Trimer Selects for Quaternary Dependent Antibodies Targeting the Trimer Apex Devin Sok, Marit J. van Gils 2, Matthias Pauthner, Karen L. Saye- Francisco, Jessica Hsueh, Bryan Briney, Jean-Philippe Julien, Peter S. Lee, Yuanzi Hua, John P. Moore 3, Ian A. Wilson, Rogier W. Sanders 2, Dennis R. Burton 4 Scripps Research Institute, La Jolla, CA, United States, 2 University of Amsterdam, Academic Medical Center, Amsterdam, Netherlands, 3 Weill Medical College of Cornell University, Department of Microbiology and Immunology, New York, CA, United States, 4 Scripps Research Institute, Department of Immunology and Microbial Science, La Jolla, CA, United States Background: Broadly neutralizing antibodies (bnabs) targeting the trimer apex of HIV envelope (Env) are favored candidates for vaccine design and immunotherapy because of their high neutralization breadth and potency. Methods to isolate bnabs against this site, however, have been limited due to the quaternary nature of the epitope region. Methods: We report the use of a recombinant HIV envelope trimer, BG505 SOSIP.664 gp40, as an affinity reagent to isolate quaternarydependent bnabs from the peripheral blood of a chronically infected donor. We chose to isolate antibodies from donor 84 from IAVI Protocol G, from whom the antibodies PGT4-45 were isolated. Results: We describe novel somatic variants of the PGT4-45 bnab family with a range of neutralization breadth and potency. Interestingly, the new variant are highly divergent from the PGT4-45 somatic variants being only 50-60% similar in amino acid sequence identity, but deriving from the same gene family and sharing similar long CDRH3s. Strikingly, variants with limited neutralization breadth were isolated that share similar overall antibody features as the broadly neutralizing variants, suggesting that differing evolutionary pathways can occur within a single donor to yield differing neutralization profiles. Conclusions: Overall, our results report for the first time the ability of a soluble protein bait to isolate quaternary-dependent antibodies. Thus, BG505 SOSIP.664 gp40 offers a useful tool for the isolation of trimerapex glycan-dependent antibodies and reveal a mosaic of antibody responses against the trimer apex within a clonal family. OA0.04 An Efficiently Cleaved HIV- Subtype C Env that Is Selectively Recognized by Neutralizing Antibodies: A Platform for Immunogen Design Saikat Boliar, Supratik Das, Manish Bansal, Brihaspati Narayan Shukla, Shilpa Patil, Tripti Shrivastava, Sandeep Goswami, C Richter King 2, Jayanta Bhattacharya, Bimal K. Chakrabarti Translational Health Science and Technology Institute, THSTI-IAVI HIV Vaccine Design Program, Gurgaon, India, 2 IAVI, IAVI Vaccine Design, New York, NY, United States Background: The HIV vaccine field continues to study the elicitation of broadly neutralizing antibodies (bnabs) targeting the Envelope glycoprotein (Env). A cleaved native HIV- Env, which selectively binds to bnabs, would be an ideal immunogen. This approach, however, has eluded researchers because of the difficulty in maintaining the structural integrity of cleaved Envs. Soluble BG SOSIP trimeric Envs (clade A) display a nearly native antigenic profile, but require mutated cleavagesite, cysteine-linkages and co-expression of furin. Priming with plasmid DNA expressing cleaved, membrane bound native Env followed by boost with soluble Env could be a feasible solution. However, JRFL (clade B) Env is the only known cleaved Env when expressed on the cell surface. Thus it is important to screen other Envs that show efficient cleavage. Methods: A total of 38 Indian subtype C env clones were screened for cleavage. Comprehensive FACS-based binding and biochemical assays were used to confirm cleavage and characterize the Env. Results: We have identified 4-2.J4Env, which is efficiently cleaved and displays similar antigenic profile to JRFL on the cell surface. A comparative analysis of both Envs show that JRFL and 4-2.J4 Envs bind to a cleavage dependent bnab, PGT5 and the cleavage-defective forms do not bind to PGT5. However, when the cytoplasmic tail (CT) is truncated, 4-2.J4delCT Env binds to both neutralizing and nonneutralizing antibodies suggesting that the cytoplasmic domain of 4-2. J4 Env plays an important role in maintaining its native conformation. Codon-optimization of the envs enhances expression but, unlike JRFL, 4-2.J4 Env demonstrates similar properties to its non-codon optimized counterpart indicating that it retains its native conformation. Conclusions: Here, we report the identification of 4-2.J4Env, an efficiently cleaved subtype C Env. As almost half of the global HIV- infections are mediated by clade C virus, this Env would advance the design and development of HIV- vaccine. 38 HIV Research for Prevention 204 HIV R4P

39 Oral Abstract Session 0: B Cell Immunogen Design Tuesday, 28 October OA0.05 Minimizing Undesirable Epitope Immunodominance on HIV- Env Immunogens through Rational Immunogen Modification Andrew T. McGuire, Anita M. Dreyer, Sara Carbonetti, Jolene A. Glenn, Johannes F. Scheid 2, Hugo Mouquet 2, Leo Stamatatos Seattle Biomed, Seattle, WA, United States, 2 The Rockefeller University, New York, NY, United States Background: Immunization with recombinant HIV- Env elicits antibodies targeting epitopes on variable regions of Env that display narrow breadth of neutralization (nnabs), but fails to elicit antibodies targeting conserved epitopes that display broad neutralizing activity (bnabs). This is thought to be in part due to the immunodominance of the variable regions of Env, as well as the inability of recombinant Env to activate naive B cells that give rise to bnabs. We recently engineered an Env capable of activating B cells expressing germline-reverted (gl) BCRs of VRC0-class bnabs and herein compare the ability of this Env to stimulate B cells expressing glvrc0-class BCRs and B cells expressing glnnab BCRs Methods: We developed novel assays to monitor the concurrent activation of B cells expressing glvrc0 class BCRs and glnnab BCRs in response to various Envs, and assays to assess Env uptake by these B cell lines. Results: Unlike B cells expressing glvrc0 class BCRs, several Envs bound to, activated, and were internalized by B cells expressing glnnab BCRs targeting both the CD4-BS and V3, indicating that nnab eptiopes are more readily presented on Env than bnab epitopes. A direct comparison of the magnitude of B cell activation responses, showed that an Env specifically engineered to engage glvrc0 class B cells, preferentially activates B cells expressing glnnab BCRs, and that the latter internalize this Env more efficiently than glvrc0 class B cells. Importantly, we demonstrate that the removal of V V2 and V3 from Env improves B cell activation and antigen uptake through glvrc0 class BCRs relative to CD4-BS and V3 directed nnab BCRs. Conclusions: Our results provide experimental evidence that explains why immunization with recombinant Env results in the production of nnabs instead of VRC0 class bnabs. However, we demonstrate that it is possible to engineer an Env-based immunogen that eliminates or reduces the immunodominance of nnab epitopes and favors the maturation process of VRC0 class bnabs. OA0.06 Uncleaved Soluble gp40 Constructs with Retained Native-like Trimer Conformation and Antigenicity Ivelin Georgiev, M. Gordon Joyce, Yongping Yang, Marie Pancera, Jason Gorman, Priyamvada Acharya, Guillaume Stewart-Jones, Aliaksandr Druz, Cheng Cheng, John R. Mascola, Peter D. Kwong National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States Background: Soluble gp40 constructs that mimic native-like HIV- Env trimers are prime immunogen candidates. gp40 cleavage into gp20 and gp4, however, is thought to be essential for mimicry of native-like conformation and antigenicity, as evidenced by binding to quaternary-specific broadly neutralizing antibodies (qbnabs) but not to non-neutralizing antibodies (NnAbs). Methods: We employed structure-based design strategies to generate modified uncleaved soluble gp40 trimers. The designed constructs were characterized using a high-throughput ELISA assay against a panel of qbnabs and NnAbs, as well as bio-layer interferometry, gel filtration and negative-stain EM. Results: The designed uncleaved gp40 constructs were initially tested in the context of the clade A HIV- strain BG505 and showed good binding to qbnabs (including highly quaternary-specific antibodies such as PGT45 and VRC26.09) but not to NnAbs (such as F05). The constructs exhibited a significant trimer fraction by gel filtration and native-like conformation by negative-stain EM. The antigenicity and conformational properties of these constructs resembled those of the current state-of-the-art soluble gp40, BG505.SOSIP, which in contrast requires full cleavage. Conclusions: Uncleaved soluble gp40 constructs that mimic nativelike trimers have the advantage of not requiring cleavage, rendering them useful candidates as protein- as well as DNA-based immunogens. Uncleaved analogs in diverse HIV- strains are currently being evaluated and immunogenicity studies are ongoing. ORAL ABSTRACT SESSIONS 39

40 Oral Abstract Sessions Oral Abstract Session 02: Microbicides: Male Partner Engagement and Sexual Behaviors ORAL ABSTRACT SESSIONS OA02.0 Engaging Male Partners in Women s Microbicide Use: Evidence from Clinical Trials and Implications for Future Research and Microbicide Introduction Michele Lanham, Rose Wilcher 2, Elizabeth T. Montgomery 3, Robert Pool 4, Sidney Schuler 5, Rachel Lenzi 5, Barbara Friedland 6, Betty Njoroge 7, Elizabeth Bukusi 7, Robyn Dayton 2 FHI 360, Social and Behavioral Health Sciences, Durham, NC, United States, 2 FHI 360, Research Utilization, Durham, NC, United States, 3 RTI International, Women s Global Health Imperative, San Francisco, CA, United States, 4 University of Amsterdam, Centre for Social Science and Global Health, Amsterdam, Netherlands, 5 FHI 360, Social and Behavioral Health Sciences, Washington, DC, United States, 6 Population Council, HIV and AIDS Program, New York, NY, United States, 7 Kenya Medical Research Institute, Research Care and Training Program, Centre for Microbiology Research, Nairobi, Kenya Background: Constructively engaging male partners in womencentered health programs such as family planning and PMTCT has resulted in both improved health and relationship outcomes. Concerted efforts to engage men in women s microbicide use for HIV prevention could make it easier for women to access and use microbicides, if an effective product is identified. Methods: We conducted primary and secondary analyses of male engagement data from six qualitative studies implemented in conjunction with microbicide trials in South Africa, Kenya, and Tanzania. The analyses included 535 interviews and 07 focus groups with trial participants, male partners, and community members. We synthesized the findings across the studies and developed recommendations for future research and microbicide introduction. Results: The majority of women in steady partnerships wanted their partner s agreement to use microbicides. Women whose male partners were resistant to microbicide use used a number of strategies to obtain their approval. Among men who were aware of their partner s microbicide use, involvement ranged from opposition to agreement/ non-interference to active support. Both men and women expressed a desire for men to have access to information about microbicides. Some women and men said that it would be helpful if male partners could talk with a health provider about microbicides; however, men were hesitant to go to the clinic during the trials because of their work schedules, fear of HIV testing, and stigma. Conclusions: We recommend counselling women on whether and how to involve their partners, providing couples counselling on microbicides, and targeting men with community education and mass media to increase their awareness and acceptance of microbicides. These activities should be tested in microbicide trials, open-label studies, and demonstration projects to identify effective male engagement approaches to include in eventual microbicide introduction. Efforts to engage men must take care not to diminish women s agency. OA02.02 Strategies to Improve Male Involvement and Partner Support in the ASPIRE Trial: The Hillbrow Experience Krishnaveni Reddy, Pranitha Ramchuran, Nombulelo Maseko, Amukelani Vuma, Lizzy Gama, Sylvia Sibeko, Nosipho Duba, Helen Rees, Thesla Palanee Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: In Southern Africa, men play a key role socially and economically in womens lives and are often more dominant in relationships. In terms of sexual health, males are less likely to exhibit health seeking behavior or access HIV/STI testing and treatment as frequently as women. This has major implications sexually and socially for women involved in reproductive health research and hence proactive male partner involvement and support in this area is crucial Methods: At Wits RHI, strategies have been implemented to enhance male involvement in the ASPIRE trial (an HIV prevention study investigating the use of intravaginal Dapivirine ring). These strategies include couple counseling, offering of facilitated trial participation disclosure sessions, hosting of male health education weekend workshops and partner invitation to study retention events. These activities, aimed at increasing reproductive/sexual health knowledge, includes information on contraception, STIs and HIV as well as ASPIRE purpose and objectives, research misconceptions and importance of partner support. Community Advisory Board members are invited to these events to facilitate discussion as needed. Results: Male partner attendance has been minimal due to work related reasons, however male partner involvement and sharing of sexual health information where possible is still encouraged at these events. Participant feedback reveals that about 50% of participants have disclosed study participation to partners with a favorable response in most cases. Partners appear supportive in terms of reminding participants of visits and product adherence with some partners being more open to HIV/STI testing and treatment. Conclusions: Male partner involvement in ASPIRE appears to increase inter-partner communication and sexual health awareness with potential to impact on study retention, product adherence and ultimately study outcomes. Therefore at Wits RHI, male involvement strategies undergo continuous evaluation and revision to optimize this aspect. 40 HIV Research for Prevention 204 HIV R4P

41 Tuesday, 28 October Oral Abstract Session 02: Microbicides: Male Partner Engagement and Sexual Behaviors OA02.03 Gel Use Disclosure in an Open-label % Tenofovir Gel Trial: Perspectives from Participants, Partners and Community Men in KwaZulu-Natal, South Africa Kathleen M. MacQueen, Sarah Dlamini 2, Brian Perry, Alesha Majors, Chitra Singh 2, Diantha Pillay 2, Eunice Okumu, Sharon Watson FHI 360, Durham, NC, United States, 2 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa Background: Disclosure of product use has been shown to influence adherence in microbicide trials. We explored disclosure in the context of CAPRISA 008, an ongoing open-label follow-on trial of % tenofovir gel enrolling uninfected CAPRISA 004 trial participants. Methods: In-depth interviews (n=63) and focus groups (n=8) were held with CAPRISA 008 participants; male partners of 3 women who fully disclosed trial participation and gel use were also interviewed. Four focus groups were held with community men who were not partners of CAPRISA 008 participants. Results: Most CAPRISA 008 participants interviewed told their partner at least some details about being in the trial or use of the gel. Motivating factors for disclosure included perceived difficulty in hiding gel from a partner and a desire to inform others about the benefits of the gel. Women disclosing gel use to a partner perceived it easier to adhere to the coitally-related regimen. Male partners of disclosers described feelings of initial fear and apprehension of the gel followed by gradual acceptance after being more fully informed. Non-disclosing women described the ease of hiding gel use from their partners. Barriers to disclosure included fear that the partner would not understand her motivations for using the gel for HIV prevention or that he would react negatively to her use of an intravaginal product. Community men saw disclosure as indicative of the quality and seriousness of a relationship: women in casual relationships could use gel without disclosing while those in stable relationships should discuss gel use because nondisclosure inferred lack of trust. Community men felt that counselors and pharmacists could assist women in explaining the purpose and use of the gel to their partner. Conclusions: Disclosure of % tenofovir gel use is perceived as beneficial by women and men when it is negotiated and socially supported. Nondisclosure is a practical and acceptable option in some relationships. OA02.04 Exploring Post-coital Intravaginal Cleansing Practices among Women Enrolled in the Microbicides Development Programme MDP 30 Clinical Trial Mitzy Gafos, Angela Crook, Robert Pool 2, Misiwe Adelaide Mzimela 3, Neetha Morar 4, Andrew Vallely 5, Andrew Abaasa 6, Thesla Palanee 7, Oliver Mweemba 8, Gita Ramjee 4, Suzanna Francis 5, Anatoli Kamali 6, Helen Rees 7, Maureen Chisembele 8, Sheena Mccormack, on behalf of the MDP Study Team MRC Clinical Trials Unit at UCL, London, United Kingdom, 2 University of Barcelona, Barcelona, Spain, 3 Zululand University, KwaDlangezwa, South Africa, 4 HIV Prevention Research Unit, Medical Research Council, Durban, South Africa, 5 Mwanza Intervention Trials Unit, National Institute for Medical Research, Mwanza, Tanzania, United Republic of, 6 MRC/UVRI Uganda Research Unit on AIDS, Kampala, Uganda, 7 Wits Reproductive Health and HIV Institute (WRHI), Johannesburg, South Africa, 8 University of Zambia, Lusaka, Zambia Background: Post-coital intravaginal cleansing (IVC) could impact the use of vaginal microbicide gels and limit the protective effect. In MDP30 participants were counselled against IVC less than hour after sex and here we report their post-coital IVC practices. Methods: We enrolled 9385 women at 3 clinics in 6 research centres: Africa Centre (AC), Durban (DU), Johannesburg (JB) in South Africa; Mwanza (MW) Tanzania, Masaka (MS) Uganda, and Mazabuka (MZ) Zambia. Data on 9096 women with follow up data are included in this analysis. Data on post-coital IVC were collected for each sex act in the last week/4 weeks, at weeks 4, 24, 40 and 52 after enrolment. Results: Despite counselling to the contrary, 42% (3,838/9096) of women reported IVC within an hour after sex at least once during the trial, mainly water or finger cleansing. IVC differed significantly across centres: 26% in JB, 29% in AC, 3% in MW, 36% in MS, 59% in DU, and 69% in MZ (p=0.00). In multivariable pooled analysis controlling for centre, IVC was associated with age < =24 (AOR.; CI.0,.22), < =primary education (AOR.6; CI.03,.30), and reporting other IV practices (AOR.70; CI.25,2,32). In multivariable centre specific analysis, IVC was associated with younger age (JB, AC), unemployment (MS), inconsistent gel use (MW), randomisation to 0.5% PRO2000 or placebo gel groups (MS, MZ), larger household size (AC), Islamic religion (MS), partner knowledge of gel (DU), other IV practices (MW) and clinic of enrolment (DU, JB, AC). Among women who reported IVC, cleansing was practiced after 43% of sex acts (on average per woman) differing by centre at 33% in JB MW, 35% in MS, 43% in AC, and 48% in DU MZ. IVC decreased from 33% in the first half to 24% in the second half of the trial, decreasing most in JB and least in MZ. Conclusions: Post-coital IVC is a common hygiene practice that may be amenable to change. We need to further understand the impact of IVC on microbicide efficacy and consider strategies to influence post-coital IVC practices. ORAL ABSTRACT SESSIONS 4

42 Oral Abstract Sessions Oral Abstract Session 02: Microbicides: Male Partner Engagement and Sexual Behaviors ORAL ABSTRACT SESSIONS OA02.05 Language, Terminology and Understanding of Anal Sex amongst VOICE Participants in Uganda, Zimbabwe and South Africa Zoe Duby,2, Miriam Hartmann 3, Imelda Mahaka 4, Elizabeth T. Montgomery 3, Christopher J. Colvin, Barbara Mensch 5, Ariane van der Straten 3 University of Cape Town, School of Public Health, Cape Town, South Africa, 2 Desmond Tutu HIV Foundation, Cape Town, South Africa, 3 RTI International, San Francisco, CA, United States, 4 UZ-UCSF Collaborative Research Programme, Harare, Zimbabwe, 5 Population Council, New York, NY, United States OA02.06 Application of a Body Map Tool to Enhance Discussion of Sexual Behaviour in Women: Experiences from MTN 003D Sarita Naidoo, Kubashni Woeber, Otilia Munaiwa 2, Juliane Etima 3, Zoe Duby 4, Miriam Hartmann 5, Elizabeth Montgomery 5, Barbara Mensch 6, Ariane van der Straten 5, and the MTN 003D Team South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 UZ-UCSF Collaborative Research Programme, Harare, Zimbabwe, 3 MU-JHU Research Collaboration, Kampala, Uganda, 4 Desmond Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town, South Africa, 5 RTI International, San Francisco, CA, United States, 6 Population Council, New York, NY, United States Background: Despite efforts to use comprehensible terms, whether research participants accurately interpret questions on sexual behaviour is not well understood. MTN-003D, a qualitative ancillary study to the VOICE HIV prevention trial in Sub-Saharan Africa, explored local vernacular for sexual behaviours, understanding of terms for penileanal intercourse (AI) in VOICE ACASI questionnaires, and participant narratives of AI behaviour compared to ACASI reports. Methods: In-depth interviews (IDIs) were conducted with 88 women (22 Ugandans, 26 Zimbabweans and 40 South Africans) in their language of preference (Zulu, Luganda, Shona or English). A diagram of a nude female figure was used as a visual aid to facilitate discussion of participant interpretations of terms for AI. Results: Findings suggest potential misreporting of AI due to ambiguity and misinterpretation of terms in VOICE ACASI questionnaires. After terms were clarified in IDIs, many participants claimed they had not understood the ACASI AI question, or that they had interpreted it to refer to vaginal sex from behind. There was also ambiguity in vernacular terms participants themselves used. Of 88 participants, 35 had reported AI in ACASI, but only 0 of these also disclosed AI during IDIs. Another 7 reported AI during the IDIs but had not done so in ACASI. Of the remaining 25 reporting AI in ACASI but not in the IDI, a third needed the term for AI to be clarified during the IDI. Conclusions: Findings highlight challenges in developing terms for sexual behaviour that are neither ambiguous nor open to misinterpretation. In efforts to balance social appropriateness with non-ambiguity across languages, terminology used in research is not always accurate or explicit. These findings suggest that some participants misunderstood the AI question in VOICE, which may have led to misreporting. The challenges in accurately reporting sexual behaviour have implications not only for clinical trials, but also for clinical practice and assessing HIV/STI risk. Background: Body maps are used in sexual and reproductive health studies to explore sex and sexuality, risks and pleasures, and to understand individual s perceptions of their bodies. MTN-003D, an exploratory study of potential sources of efficacy dilution in the VOICE trial, used a body map tool, together with in-depth interviews (IDIs), to encourage frank discussion about sexual behaviour. Methods: IDIs were conducted with 88 former VOICE participants in Zimbabwe, Uganda and South Africa from December 202 to March 203. Body map tools, which showed the front and back outline of a nude female figure, were used during IDIs to initiate and aid discussion of sex. Interviewers asked women to identify and discuss genitalia and other body parts associated with sexual behaviour, pain and pleasure; and their responses (key words or short sentences) were noted on the map. Analysis of annotated body maps was done together with corresponding transcript data to ascertain the success of this tool. Results: Body maps helped interviewers probe issues around sexual behaviour. Women could point to body parts without having to verbalise potentially embarrassing anatomical terms. Women were amused, shy or embarrassed when shown the map. Most women readily labelled maps; however some needed clarity or encouragement. Body maps were used by most women to discuss pleasure, however only half of the women used the tool to identify areas associated with pain. Only 6 women were reluctant to discuss their sexual behaviour (2 refused due to religious beliefs; only labelled biological functions of the body; 2 avoided looking at map; uncomfortable to discuss anal sex). Conclusions: In this study, body maps used in combination with IDIs were feasible and effective tools to enhance discussions around sexual behaviour, specifically anal sex. Body maps provided women with a non-intimidating way of discussing and disclosing their sexual practises and minimized miscommunication of anatomical terminology. 42 HIV Research for Prevention 204 HIV R4P

43 Tuesday, 28 October Oral Abstract Session 03: Animal Model Studies of Microbicides and Injectables OA03.0 Preclinical Evaluation of TMC-278 LA, a Long-acting Formulation of Rilpivirine, Demonstrates Significant Protection from Vaginal HIV Infection Olivia Snyder, Heather Vincent, Sophie Lachau-Durant 2, Guenter Kraus 2, Peter Williams 2, J. Victor Garcia UNC Chapel Hill, Chapel Hill, NC, United States, 2 Janssen Research and Development, Beerse, Belgium Background: Vaginal HIV transmission accounts for the majority of new infections worldwide. Prevention efforts have demonstrated mixed success due to lack of adherence to drug regimens. New efforts to prevent HIV transmission have focused on long acting (LA) antiretroviral drug formulations to circumvent this issue. Methods: We first established critical pharmacokinetic parameters of intramuscular (IM) injection of TMC 278 LA in mice and demonstrated sustained drug release for 4 weeks. Humanized BLT mice were then vaginally challenged with HIV transmitted founder viruses (T/F) after IM administration of TMC 278 LA and the extent of protection from HIV acquisition was determined. In the st experiment, challenges with the T/F virus CHO40 were performed week after drug administration (600mg/kg). In a 2nd experiment, BLT mice were challenged week after drug administration with of 3 different viruses (CHO40, RHPA, or JR-CSF). These mice were exposed to a second challenge 3 weeks after drug administration with a different T/F virus (THRO). Infection was determined using viral load assay and PCR analysis for vdna in tissues. Identity of the infecting viruses was confirmed by DNA sequencing. Results: In the first experiment, a single IM dose of TMC 278 LA (600 mg/kg) one week before vaginal challenge provided significant protection from CHO40 infection (6/6 mice protected) (p=0.0047). In contrast, 3/3 animals that received saline became infected. In the second experiment, 6/7 BLT mice that received saline before exposure became infected. Whereas 6/8 BLT mice that received TMC-278 LA before the first viral challenge were protected from infection (p=0.026). All mice exposed 3 weeks post drug administration to a 2nd challenge with THRO became infected. Conclusions: TMC-278 LA offers significant protection from vaginal HIV infection against T/F viruses. Although a wane in protection over time was observed. These results demonstrate the potential of long acting antiretroviral formulations for HIV prevention. OA03.02 LB HIV PrEP Dose Rationale for Cabotegravir (GSK265744) Long-acting Injectable Nanosuspension Bill Spreen, Alex Rinehart 2, Kimberly Smith 2, David Margolis, Susan Ford 3, Steve Piscitelli GlaxoSmithKline R&D, Infectious Diseases, Research Triangle Park, NC, United States, 2 ViiV Healthcare, R&D, Research Triangle Park, NC, United States, 3 GlaxoSmithKline R&D, Clinical Pharmacology Modeling and Simulation, Research Triangle Park, NC, United States Background: Long acting (LA) drugs such as the integrase inhibitor cabotegravir (CAB, GSK265744) may bolster PrEP adherence and efficacy. PrEP dose selection is challenging given lack of validated surrogate markers and low rates of HIV seroconversion. Methods: CAB PrEP dose selection was based on in vitro virology, human antiviral activity, healthy volunteer (HV) single and repeat dose safety and PK studies and non-human primate (NHP) PK and SHIV rectal/vaginal challenge studies. Data were integrated in populationbased PK (PPK) analyses/dose simulations to identify a dose predicted to be efficacious and clinically practical. Results: In a 0-day monotherapy study in ART-naïve patients CAB 5mg/ day p.o. (n=7) yielded geomean blood plasma (BP) C tau of 0.57 ug/ml, >3-fold above protein-adjusted (PA)-IC 90 of 0.66 ug/ml, and produced a 2.4 log median decrease in HIV RNA, providing a PK/PD correlate. Single injection CAB LA (SC or IM) in 58 HV evaluated ascending doses for safety and PK. CAB LA 800mg IM BP levels exceeded C tau of oral 5mg/day for 6 weeks. A repeat dose HV (n=40) study confirmed safety and PK of 800mg IM dose; geomean C tau (week 2) was. ug/ml or 7x PA-IC 90. NHP PK studies targeting CAB concentration-time profiles in BP approximating human values were followed by SHIV62p3 rectal challenges (50xTCID 50 ) to establish POC and evaluate threshold of protection. CAB BP xpaic 90 correlated with 97% protection. SHIV vaginal challenge showed protection in 6/8 rhesus macaques (300xTCID 50 ) and 8/8 pigtail macaques (50xTCID 50 ). PPK based simulation of dosing regimens achieving target BP levels with quarterly injections enabled selection of 800mg IM q2week dose. Conclusions: CAB LA PrEP dose selection was guided by human safety, PK and antiviral activity studies and NHP challenge studies. CAB LA 800 mg IM q2weeks is predicted to yield drug levels correlated with robust anti-hiv activity and protection against SHIV rectal and vaginal challenge. This dose is under study in phase 2 safety and PK trials. ORAL ABSTRACT SESSIONS 43

44 Oral Abstract Sessions Oral Abstract Session 03: Animal Model Studies of Microbicides and Injectables ORAL ABSTRACT SESSIONS OA03.03 Tenofovir Reservoir Intravaginal Rings Provide Superior Pharmacokinetics and Higher Sustained Drug Levels than Tenofovir Matrix Rings Meredith Clark, Lara Pereira 2, Justin Clark 3, Todd Johnson 3, Chou-Pong Pau 4, David Friend, James Smith 4, Patrick Kiser 5 CONRAD Eastern Virginia Medical School, Arlington, VA, United States, 2 LifeSource Biomedical LLC, Moffett Field, CA, United States, 3 University of Utah, Salt Lake City, UT, United States, 4 Centers for Disease Control and Prevention, Division of HIV and AIDS Prevention, Atlanta, GA, United States, 5 Northwestern University, Evanston, IL, United States OA03.04 A Combination Vaginal Ring Releasing Dapivirine and Darunavir Karl Malcolm, Diarmaid Murphy, Delphine Desjardins 2,3, Nathalie Dereuddre-Bosquet 2,3, Patricia Brochard 2,3, Ludivine Perrot 2,3, Alain Pruvost 4, Roger Le Grand 2,3, Ole Lagatie 5, Leen Vanhooren 5, Maxim Feyaerts 5, Jens Van Roey 5 Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 Commissariat à l Energie Atomique (CEA), Division of Immuno-Virology, DSV/iMETI, IDMIT Center, Fontenay-aux-Roses, France, 3 Paris Sud University-, UMR-E, Orsay, France, 4 Commissariat à l Energie Atomique (CEA), ibitecs, SPI, Laboratoire d Etude du Métabolisme des Médicaments, F-99, Gif-sur-Yvette, France, 5 Janssen Diagnostics, Turnhoutseweg, Belgium Background: Sub-protective topical antiretroviral (ARV) levels can lead to HIV transmission events. Therefore maintaining drug levels in tissues is a primary goal of long acting ARV delivery systems. Matrix intravaginal rings (IVR) show attenuating release rates over time; reservoir IVR can deliver precise drug levels over several months. Both IVR types are being investigated in the topical PrEP field. Here we present the in vitro and in vivo evaluation of TFV release and pharmacokinetics (PK) of matrix and reservoir polyurethane IVR in pigtail macaques. Methods: Macaque-sized IVR contained 360mg TFV in a solid hydrophilic polyurethane (HPU) matrix or 550mg TFV in a hollow-core HPU reservoir. IVR were characterized for in vitro release. IVR were inserted vaginally into pigtail macaques (N=4-6) for 28d; blood plasma, vaginal secretions (VS), rectal secretions (RS), vaginal tissue (VT) and rectal tissue (RT) biopsies were collected at various time points. TFV PK was assessed by LC/MS-MS. Results: Matrix IVR demonstrated time-dependent TFV release and PK profiles, with ~-log reductions observed for both in vitro release rate (32 to 4 mg/d) and median TFV concentrations in VS (3-6x0 6 to 3-4x0 5 ng/ml), VT (0.9-3x0 4 to 0.6-x0 3 ng/g), and RT (497 to 76.5 ng/g) over the 28d treatment period. Median TFV RS levels were sustained at < IC 50 levels ( ng/ml). Reservoir IVR showed time-independent TFV release and PK profiles over the same duration. In vitro release was ~6mg/d TFV, and median TFV levels were sustained at 2-7x0 6 ng/g (VS), x0 5 ng/g (VT), and 0.2-.x0 4 ng/g (RS). Conclusions: TFV reservoir IVR provide superior PK control and higher sustained vaginal TFV levels compared to TFV matrix IVR in macaques. Previous non-human primate studies that evaluated TFV % gel and tenofovir disoproxil fumarate IVR suggest these local concentrations are likely to be protective from vaginal challenge in the macaque model. Background: Combination microbicide vaginal rings, containing two or more antiretrovirals targeting different steps in the HIV replicative process, may be more effective than single microbicide products at preventing sexual transmission of HIV. Here, we report the preclinical development, including in vitro release and macaque pharmacokinetics, of matrix-type silicone elastomer rings containing dapivirine (DPV; an experimental non-nucleoside reverse transcriptase inhibitor) and darunavir (DRV; a marketed protease inhibitor). Methods: Macaque rings containing 25 mg DPV, 300 mg DRV and 00 mg DPV, and 300 mg DRV were manufactured and characterised by differential scanning calorimetry. In vitro release was assessed into isopropanol/water and simulated vaginal fluid. Macaque vaginal fluid and blood serum concentrations for both antiretrovirals were measured during 28-day ring use. Tissue levels were measured on day 28. Ex vivo challenge studies were performed on vaginal fluid samples and IC50 values calculated. Results: DRV caused a concentration-dependent reduction in the DPV melting temperature in both solid drug mixes and in the combination ring. In vitro release from rings was dependent on drug loading, the number of drugs present, and the release medium. In macaques, serum concentrations of both microbicides were maintained between pg/ml. Vaginal fluid levels ranged between ng/g and ng/g for DPV and DRV, respectively. Vaginal tissue concentrations decreased in rank order: vagina ( ng/g) > cervix ( ng/g) > uterus (0-08 ng/g) > rectum (0-40 ng/g). Measured IC50 values (HIV- BaL) determined from macaque vaginal fluid samples were < 2 ng/ml for both compounds. Conclusions: Based on these results, and in light of the ongoing clinical progress of the 25mg DPV ring, a combination vaginal ring containing DPV and DRV is a viable second-generation HIV microbicide candidate. 44 HIV Research for Prevention 204 HIV R4P

45 Tuesday, 28 October Oral Abstract Session 03: Animal Model Studies of Microbicides and Injectables OA03.05 A Novel Intravaginal Ring (IVR) Protects Macaques against SHIV-RT Infection and Reduces HSV-2 Shedding after Repeated SHIV-RT/HSV-2 Co-challenge Thomas M. Zydowsky, Jessica Kenney, Meropi Aravantinou, Shweta Ugaonkar, Nina Derby, Larisa Kizima, Shimin Zhang, Olga Mizenina, Jose Fernández-Romero, Melissa Robbiani Population Council, HIV & AIDS Program, New York, NY, United States Background: MZC gel (MIV-50, zinc acetate [ZA], carrageenan [CG]) significantly reduces vaginal SHIV-RT and HSV-2 infection in macaques when given 8h before challenge. CG gels significantly reduce HPV infection in mice when dosed 24h before challenge. We aim to develop a 90d IVR releasing MZC and levonorgestrel (LNG) to prevent HIV, HSV- 2, HPV, and conception. Methods: IVRs contained 3mg MIV-50±0.6mg LNG in the matrix and 30mg ZA/70mg CG in the core (open to fluids via a matrix pore). We measured CG, MIV-50, and LNG in macaque blood and/or vaginal swabs during and after 28d of IVR insertion. For efficacy, we exchanged IVRs every 2d, challenging macaques with 200 TCID 50 SHIV-RT/0 7 pfu HSV-2 on d7, 0, 4 and 7 post IVR insertion (20 challenges total); n=4 placebo IVR, n=4 LNG IVR, n=2 MZC IVR, n=2 MZCL IVR. Results: MIV-50 was detected in swabs and plasma h post IVR insertion, peaked at d in swabs and d-3 in plasma, declined from d7- d28, and was undetectable 24h after IVR removal. LNG was detected in serum within 4h, plateaued by d7, and was undetectable 24h after IVR removal. CG was detected in swabs of most animals from d3 until the IVRs were removed. Preliminary data show MZCL IVRs protected (92%) macaques against SHIV-RT (/2 vs. 4/4 infected in LNG IVR controls; p=0.003); 2/2 MZC and 2/4 placebo IVR animals became SHIV-RT infected (67% protection). MZC and MZCL IVRs significantly protected compared to control IVRs (3/24 vs. 6/8 infected, p=0.002). Initial data suggest that ~30% fewer animals became infected with HSV-2 after treatment with MZC and MZCL IVRs (7/24 vs. 8/8 infected controls); the frequency (8/30 vs. 23/95 time points positive; p< ) and levels (67/80 vs. 3/570 replicates positive; p< 0.000) of HSV-2 shedding were significantly lower than in the controls. Conclusions: We developed an IVR that releases APIs targeting HIV, HSV-2, HPV, and conception. The IVR significantly reduces SHIV-RT infection and HSV-2 shedding in macaques. OA03.06 LB Rectal Specific Gels Containing Maraviroc and/or Tenofovir Protect against Rectal SHIV Transmission in a Macaque Model Charles Dobard, Andrew Taylor, Sunita Sharma, Dinh Chuong, Chou-Pong Pau, Lisa Rohan 2, Ian McGowan 2, Walid Heneine Centers for Disease Control and Prevention, Atlanta, GA, United States, 2 Magee Womens Research Institute, University of Pittsburgh, Pittsburgh, PA, United States Background: Rectal transmission of HIV is an important driver of the HIV epidemic in several populations. Rectal microbicides containing antiretroviral drugs are under development to prevent rectal acquisition of HIV. Maraviroc (MVC) and tenofovir (TFV) are candidate drugs for rectal-specific gel formulations. Previous work showed that gels formulated for vaginal application were not optimal for rectal use. Here we evaluated the efficacy of optimally formulated rectal gels containing % MVC, % TFV, or in combination against repeated rectal SHIV exposures in rhesus macaques. Methods: MVC and TFV were formulated in a rectal-specific iso-osmolar hydrogel based formulation at neutral ph. Macaques were administered 4 ml of % MVC (n=6), % TFV (n=6), %MVC/%TFV (n=6) or a placebo (n=7) gel 30 minutes before each rectal SHIV SF62P3 challenge (500 TCID 50 ). Challenges were repeated twice-weekly for 5 weeks (0 challenges). Plasma drug levels were measured 30 min after gel application using LC/MS/MS. Infection was monitored by serology and PCR of SHIV in plasma. Infected macaques continued to receive gel for an additional 6-weeks to monitor impact on systemic viremia. Results: All 7 controls were infected after a median of 4 challenges. In contrast, 4 of 6 macaques in each group receiving either % MVC, % TFV, or MVC/TFV gel remained protected after 0 challenges demonstrating an 83% efficacy (p=0.02). Low levels of MVC (median=4 ng/ml) and TFV (median=9 ng/ml) were detected in plasma 30 minutes after rectal dosing, suggesting rapid absorption. There was no difference in drug plasma concentrations between protected and breakthrough infections. Similar plasma viremia in controls and breakthroughs was observed reflecting the low systemic drug exposure. Conclusions: Here we demonstrated in vivo drug release by 3 gel formulations and showed that all provided high efficacy against repeated rectal SHIV exposures. Our findings support the clinical development of these gels for rectal prophylaxis against HIV acquisition. ORAL ABSTRACT SESSIONS 45

46 Oral Abstract Sessions Oral Abstract Session 04: Innate Immunity ORAL ABSTRACT SESSIONS OA04.0 SIV- and Vaccine-elicited NK Cell Memory in Rhesus Macaques R. Keith Reeves,2, Haiying Li,2, Eryn Blass, Hualin Li, Stephanie Jost 3, Marcus Altfeld 3, Dan Barouch Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, United States, 2 New England Primate Research Center, Southborough, MA, United States, 3 Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States Background: Natural killer (NK) cells provide rapid responses to viral infections and are typically considered to be nonspecific components of innate immunity. However, recent studies have shown that NK cells can also mediate antigen-specific memory in mice, but it remains unclear whether this phenomenon also exists in primates. Methods: In this study we evaluated NK cells from a cohort of 8 rhesus macaques chronically infected with SIVmac25, 6 naïve controls, and 9 macaques vaccinated with replication-incompetent Ad26 vectors expressing either HIV- Env or SIVmac239 Gag. Using a novel flow cytometric assay we evaluated antigen-specific killing of autologous dendritic cells (DCs) by highly purified hepatic and splenic NK cells. Fluorochrome-labeled DCs were pulsed with intact SIV Gag or HIV- Env and non-pulsed DCs served as intra-well controls. Purified NK cells were co-cultured with DCs at multiple E:T ratios and specific lysis was used as a functional determination of antigen-specificity. Results: Splenic NK cells from SIV-infected animals were highly reactive to Gag-pulsed DCs at a 0: NK:target ratio with a median specific lysis of 40% compared to % in naive controls, indicating the presence of antigen-specific NK cells in chronic infection. In our vaccinated, unchallenged macaques, at both 0: and 5: NK:target ratios, splenic and hepatic NK cells lysed antigen-matched targets at higher frequencies than they did antigen-mismatched targets (P = 0.02, liver; P = 0.040, spleen). Responses in peripheral blood were marginal, suggesting that memory NK cells likely reside in tissues. Conclusions: Taken together, our data demonstrate the first evidence of NK cell memory in a primate species. Furthermore, antigen-specific NK cell responses to SIV antigens are induced in primates following both infection and vaccination. The longevity, functionality, and specificity of memory NK responses in primates suggest their functional relevance in developing vaccines against multiple pathogens, including HIV. OA04.02 Innate Lymphoid Cells are Depleted in HIV Infection Henrik N. Kløverpris,2, Aslam Noorbhai 3, Warren Kuhn 4, Marisa Yadon, Duran Ramsuran, Sheperd Nhamoyebonde, Victoria Kasprowicz, Bruce Walker 5, Thumbi Ndung u, Philip Goulder 6, Abdool S. Karim 7, Jenny Mjösberg 8, Alasdair Leslie KwaZulu-Natal Research Institute for TB&HIV, K-RITH, Durban, South Africa, 2 University of Copenhagen, Dept. of Int. Health, Immunology and Microbiology, Copenhagen, Denmark, 3 University of KwaZulu- Natal, Dept of General Surgery, King Edward Hospital, Durban, South Africa, 4 University of KwaZulu-Natal, Dept of ENT, King Edward Hospital, Durban, South Africa, 5 Ragon Institute, Massachusetts Institute of Technology and Harvard, Cambridge, Boston, MA, United States, 6 Oxford University, Paediatrics, Oxford, United Kingdom, 7 Center for the AIDS Programme of Research in South Africa, CAPRISA, Durban, South Africa, 8 Karolinska Institutet, Center for Infectious Medicine, Stockholm, Sweden Background: Innate Lymphoid Cells (ILCs) lack rearranged antigen receptors but share functional and developmental characteristics with lymphocytes and can be subdivided into three main groups according to their production of Th, -Th2 and -Th7 cell-associated cytokines named ILC, ILC2 and ILC3s, respectively. ILCs play a crucial role in tissue homeostasis and repair in both non-infectious and infectious diseases. HIV- pathology involves lymphoid tissue destruction of the gut mucosa associated with microbial translocation, immune activation and disease progression in both ARV-treated and untreated individuals. We hypothesized that HIV modulates the ILC population in blood and tissue. Methods: We used phenotypic and transcriptional characterization of human ILCs from a total of n=83 chronic HIV-infected and n=64 HIV uninfected individuals recruited from 4 different cohorts in Durban, South Africa. Results: We show that the Th2 cytokine producing ILC2 population in human peripheral blood is severely depleted in viremic HIV- infected individuals (P< 0.000) but preserved in those with natural immune control of HIV- (P=0.003), and correlates negatively with plasma viral load (r=-0.62, P=0.0004). This ILC2 population is depleted from peripheral blood within 20 days of HIV- transmission (P=0.04) and only partially restored upon successful antiretroviral therapy in a subset of individuals. Similar depletion of ILC and ILC3 populations were observed in chronic infection (P< 0.00) to similar kinetics as the ILC2 population, but distinct from the observed NK cell expansion dynamics. Preliminary data from gut and tonsil tissue resident cells shows overall reduction of ILCs skewed towards an NCR3 + ILC3 population. Conclusions: These data demonstrate that HIV- infection rapidly and profoundly modulates the tissue resident and circulating ILC population, with great potential significance to the pathology of this disease. 46 HIV Research for Prevention 204 HIV R4P

47 Oral Abstract Session 04: Innate Immunity Tuesday, 28 October OA04.03 Impact of Systemic Immune Activation (IA) and Inflammation on the HIV Susceptibility of HIV- individuals with HIV Concordant or Discordant Partners Shameem Z. Jaumdally,2, Pamela P. Gumbi, Hoyam Gamieldien, Lindi Masson, Heather B. Jaspan 3,4, Caroline Tiemessen 5,6, Anabela Picton 5,6, Anna-Lise Williamson,7, David Coetzee 8, Francesca Little 9, Jo-Ann S. Passmore,7 University of Cape Town, Medical Virology, Cape Town, South Africa, 2 University of Cape Town, Department of Public Health and Family Medicine, Cape Town, South Africa, 3 University of Cape Town, Immunology, Cape Town, South Africa, 4 Seattle Biomedical Research Institute, Seattle, WA, United States, 5 National Institute for Communicable Diseases of the NHLS, Johannesburg, South Africa, 6 Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 7 National Health Laboratory Service, Groote Schuur Hospital, Cape Town, South Africa, 8 Department of Public Health and Family Medicine, Cape Town, South Africa, 9 University of Cape Town, Department of Statistical Science, University of Cape Town, Cape Town, South Africa Background: Studies of individuals who appear to resist HIV infection, such as HIV- partners in HIV discordant relationships, are important for identifying host responses or characteristics associated with protection against HIV infection. Previously, immune quiescence has been associated with HIV resistance. The aim of this study was to compare the level of IA and systemic inflammation in HIV- individuals from South Africa (SA) who were either in relationships with HIV-infected or uninfected stable partners, to evaluate markers of HIV exposure or resistance. Methods: A heterosexual couples cohort of 03 HIV- individuals with long-term stable HIV- concordant partners (HIV- unexposed) and 3 HIV- individuals with HIV+ discordant partners (HIV- exposed) were included in this study. T cell activation and proliferation (CD38, HLA- DR, CCR5, Ki67) in blood was assessed by flow cytometry. Cytokines in plasma were evaluated by Luminex. Results: HIV- exposed individuals had lower frequencies of CD4+ T-cells in blood expressing the CCR5 [alone (p=0.05) or in combination with Ki67 (p=0.05) or CD38 (p=0.05)] than HIV-unexposed individuals. Similarly, HIV- exposed individuals had significantly lower frequencies of CD8+ T-cells in blood expressing CCR5 [alone (p=0.05) or in combination with Ki67 (p=0.0) and CD38 (p=0.05)] and HLA-DR (p=0.0) than their HIV- unexposed counterparts. Plasma concentrations of IL-2 (p=0.02), IFN-γ (p=0.05) and GM-CSF (p=0.006, stayed sig. after adjustment for multiple comparisons) were significantly lower in HIVexposed compared to HIV- unexposed individuals. Conclusions: This study suggests that HIV- exposed individuals from SA have an immune quiescent phenotype, with lower frequencies of activated CCR5-expressing T-cells and CCR5 density per T cell, than their HIV- unexposed counterparts. Since CCR5 expressing T-cells, especially activated ones, are the preferred targets for HIV infection, this study in HIV- discordant couples suggests that CCR5 agonists may be useful to block HIV infection. OA04.04 Do CD6+ NKG2A+ NK Cells Recruited to the Gut Combined with Passively Administered SIV Specific Antibodies Prevent SIV mac25 Acquisition in Macaques? Namal P.M. Liyanage, Melvin N. Doster, Francesca Caccuri, Poonam Pegu, Shari N. Gordon, Robyn Washington Parks, Cynthia Pise-Masison, Luis Barcena 2, Jeff Scheider 2, Howard Y. Lakouga 2, Patrick Kiser 2, Stephen Whitney 3, Luca Schifanella, Monica Vaccari, Thomas J. Hope 2, Genoveffa Franchini National Cancer Institute, NIH, Animal Models & Retroviral Vaccine Section, Bethesda, MD, United States, 2 Northwestern University- Feinberg School of Medicine, Robert H Lurie Medical Research Center 303 E Superior, Chicago, IL, United States, 3 Advanced BioScience Laboratories, Inc., Rockville, MD, United States Background: Growing evidence suggests that NK cells and antibodies that mediate ADCC can control HIV infection. ADCC was shown to have an inverse correlation with risk of HIV infection in the RV44 vaccine trial that showed 3% protection from HIV acquisition. We recently showed the recruitment of CD6+ NKG2A+ NK cells to the gut during ALVAC/SIV/gp20 vaccine regimen in macaques. Here we investigated whether NKG2A+ NK cells together with vaccine induced anti-envelop IgG at mucosa can protect from SIV mac25 acquisition. Methods: IgG was purified from sera following 8 additional immunizations with ALVAC-SIV/gp20/Alum of 8 macaques previously exposed to SIVmac 25 remaining uninfected. To demonstrate that systemically given IgG can localize to the gut, CY5 labeled antihuman IgG was subcutaneously administered to macaques and antihuman antibodies were measured in serum and rectum by ELISA and immunohistochemistry (IHC) at 6h, 24h and weekly for 4 weeks. Two macaque cohorts (A and B) were vaccinated with ALVAC-SIV (SIV 766 Gag-Pro gp20tm) at 0, 4, 2 and 24 weeks. ALUM was administered together with the vaccine at 2 and 24 weeks. Purified IgG will be given to group A (n=8) subcutaneously (24h) and intra-rectally (IR) (2h) prior to the repeated low dose challenge with SIV mac25 IR in July 204. Group B (n=8) and unvaccinated group (n=2) will be challenged simultaneously. Results: Purified IgG from 8 protected animals showed.87% of gp20 specific activity by ELISA. Anti-human IgG was detected in serum and rectal biopsies by ELISA and IHC up to 3 weeks post antibody administration. Cytometric analysis of rectal biopsies showed recruitment of NKG2A+NK (p=0.03) cells to the gut of vaccinees. Conclusions: Our current data shows the recruitment of NKG2A+ NK cells to the gut and successful delivery of antibodies to the gut mucosa. Protection after SIV challenge in the end of July 204, will confirm our hypotheses that the vaccine mediated recruitment of NK cell together with antibodies can prevent SIV acquisition. ORAL ABSTRACT SESSIONS 47

48 Oral Abstract Sessions Oral Abstract Session 04: Innate Immunity ORAL ABSTRACT SESSIONS OA04.05 LB Integrated Systems Biology Analysis Reveals Contrasting Role for Innate Immune Response Genes in Conferring Risk of Infection in RV44 Trial Ali Filali-Mouhim, Slim Fourati, Francois Lefebvre, Rasmi Thomas 2, Raphael Gottardo 3, Nicole Frahm 3, Stephen De Rosa 3, Peter Wilkinson, Petra Stafova, Amit Sabnis, Jaranit Kaewkungwal 4, Punnee Pitisuttithum 4, Sorachai Nitayanphan 5, Supachai Rerks-Ngarm 6, Nelson Michael 7, Jerome Kim 2, Merlin L. Robb 2, Robert J. O Connell 8, M. Juliana McElrath 3, Mark Cameron, Rafick Sekaly 9 Vaccine & Gene Therapy Institute of Florida, Port Saint-Lucie, FL, United States, 2 U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Bethesda, MD, United States, 3 Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand, 5 Royal Thai Army, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 6 Department of Disease Control, Ministry of Public Health, Nonthaburi, Thailand, 7 U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Bethesda, MD, United States, 8 U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 9 Department of Pathology Case Western Reserve University, Cleveland, OH, United States Background: The RV44 trial showed promise towards the development of an effective HIV vaccine. Antibodies to HIV Env VV2 loop, Envspecific IgA and HLA class II DQB*06 allele with high Env IgA were all significantly associated with risk of infection. Methods: We used an integrated systems biology approach to identify novel correlates of immunogenicity and/or protection. Env-specific stimulated peripheral blood mononuclear cells from RV44 participants were used to test the hypothesis that innate pro-inflammatory responses would demarcate RV44 case-control groups. Results: A case/control gene expression analysis using 33 controls and 27 cases yielded few significantly differentially expressed genes between cases and controls. To adjust for the heterogeneity and the case/control class imbalance, we employed a stratification strategy using a treebased classification method with IgA, VV2 and DQB*06 as predictor variables and the infection status as the outcome. The obtained tree model showed a balanced accuracy of 6% and identified two different risk groups. Cases in the first group had low IgA titers and did not express the DQB*06 allele. Transcriptional profiling showed downregulation of the type II interferon induced genes (Fisher enrichment test p value =0.04) in these cases compared to controls. Cases in the second Group had high IgA and Low VV2 titers. Transcriptional profiling showed, interestingly, up-regulation of several type I interferon genes including antiviral genes (Fisher enrichment test p value =0-6 ) in these cases compared to controls. Conclusions: Herein we have identified a gene expression signature, segregating case and control donors and which, counterintuitively, shows contrasting roles for innate immune response genes in conferring vaccine induced protection. These results have important implications for the development of more effective HIV vaccine strategies. OA04.06 Loss of Viral Control in a Subset of HIV- Infected Long-term Non-progressors Is Associated with a Decline of an Array of Antiviral Responses Mohamed El-Far, Pascale Kouassi, Odalis Asin-Milan, Annie Chamberland, Mohamed Sylla, Jean-Philippe Goulet, Petronela Ancuta, Jean-Pierre Routy 2, Danielle Rouleau, Marianne Harris 3, Nicole Bernard 2, Cécile Tremblay Centre de Recherche du CHUM, Montreal, QC, Canada, 2 McGill University, Montreal, QC, Canada, 3 BC Centre for Excellence in HIV/ AIDS, Vancouver, BC, Canada Background: Understanding molecular mechanisms of natural disease control in the absence of treatment in a minor group of HIV-infected subjects, identified as slow progressors (SP), represents a logical approach towards a potential functional cure. Methods: In the current study we aimed to identify molecular signatures associated with viral control in our Canadian cohort of HIV-infected SP (Study # CTN 247; subjects maintaining CD4 counts over 500/microliter for over 7 years, in the absence of treatment). To this end, we have identified 5 subjects that have experienced a sudden loss of viral control (average increase of viral load: 5 to 79 fold) and a decline in the absolute CD4 counts (average loss: 2 cells/microliter). We used the Illumina technology to study genome-wide transcriptional profiles in peripheral blood mononucleated cells (PBMCs) isolated from these subjects before (Visit ) and after (Visit 2) the loss of virological control. Results: Our analysis identified,38 probe sets corresponding to,268 genes that were differentially expressed between V and V2 (nominal p-value < 5%). Among these genes, 864 were downregulated and 57 were upregulated at V2 versus V. Of note, among the downregulated genes, several members of both innate and adaptive anti-viral responses were identified such as APOBEC3G, IL-32, the T cell receptor CD96 (known to associated with the superior quality of CD8 T cells in HIV-infected Elite controllers), Lck, LAT, JAK, ITK and the IL-7 receptor (IL-7R). Our validation assays are currently in progress to validate the expression of these genes at the protein level. Conclusions: Our current study takes advantage of the power of the whole genome transcriptional analysis on privileged samples from HIVinfected SP subjects before and after the loss of virological control to identify new targets for therapeutic interventions and also to identify early biomarkers to predict disease progression. 48 HIV Research for Prevention 204 HIV R4P

49 Oral Abstract Session 05: Vaccine, Viral Latency and Cure Tuesday, 28 October OA05.0 Durable Suppression of Established Transmitted Founder Replication in Infected BLT Humanized Mice by Vectored ImmunoTherapy Cailin Deal, Yong Ouyang 2, Christin M. Hong, Dong Sung An 3, David Baltimore 2, Alejandro B. Balazs Harvard University, The Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 California Institute of Technology, Biology, Pasadena, CA, United States, 3 University of California at Los Angeles, Department of Translational Sciences, Los Angeles, CA, United States Background: Recent reports in humanized mice and monkeys have found that broadly neutralizing antibodies (bnabs) can suppress the replication of laboratory strains of HIV and SHIV while bnab concentration remains high. Vectored ImmunoProphylaxis (VIP) results in long-lived bnab expression following a single intramuscular (IM) injection of a specialized viral vector, and this approach has been demonstrated as a means of durably suppressing viral load. However, previous reports of VIP-delivered bnabs for HIV therapy required prior antiretroviral drug therapy to reduce viral load to prevent escape. Methods: Humanized BLT mice were infected IV with the REJO.c transmitted molecular founder strain of HIV. A low dose of combination antiretroviral therapy (ART) was administered to these animals for 5 weeks, followed by a single IM injection of VIP expressing VRC07 or luciferase. Mouse plasma was analyzed by ELISA to determine antibody concentration and by qpcr to determine viral load. Cellular fractions were analyzed by flow cytometry to quantify human CD4 cells over time. After sacrifice, plasma was subjected to a clinically validated ultrasensitive PCR-based viral load assay. Results: We detected viral loads of 0 5 copies/ml in infected mice prior to low-dose ART treatment, which resulted in a transient reduction and rebound to pre-therapy loads. Following VIP administration, we observed a rapid increase in the blood concentration of VRC07. Mice expressing VRC07 exhibited a sharp decline in viral load to undetectable levels and an increase in CD4 cells over four weeks and this effect was sustained for the remaining 8 weeks of the study. In contrast, mice expressing luciferase exhibited increasing viral loads with concomitant decreases in CD4 cells throughout the study. Conclusions: Our results demonstrate that VIP expressing VRC07 is sufficient to suppress actively replicating transmitted founder virus at high viral load and support efforts to move Vectored ImmunoTherapy into clinical trials with infected patients. OA05.02 An HIV DNA Vaccine Delivered by Electroporation and Boosted by rvsv HIV- Gag Is Safe and Immunogenic in Healthy HIVuninfected Adults Christine M. Hay, Gregory J. Wilson 2, Marnie Elizaga 3, Sue Li 4, Nidhi Kochar 4, Mary Allen 5, Magdalena Sobieszczyk 6, Ian Frank 7, Nicole Frahm 3, Georgia D. Tomaras 8, Michael Egan 9, Michael Pensiero 5, John Eldridge 9, NIAID HIV Vaccine Trials Network University of Rochester Medical Center, Rochester, NY, United States, 2 Vanderbilt University, Nashville, TN, United States, 3 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 SCHARP - FHCRC, Seattle, WA, United States, 5 DAIDS/NIAID/NIH, Rockville, MD, United States, 6 Columbia University, New York, NY, United States, 7 University of Pennsylvania, Philadelphia, NY, United States, 8 Duke University, Durham, NC, United States, 9 Profectus Biosciences, Tarrytown, NY, United States Background: Eliciting HIV-specific immune responses through vaccination remains an important goal in preventing HIV. Here we present safety, tolerability, and immunogenicity data from a phase Ia trial of a novel HIV- multi-antigen (MAG) DNA vaccine delivered by electroporation (EP) with DNA IL-2 adjuvant and boosted with an rvsv HIV- Gag vaccine. Methods: HVTN 087 enrolled 00 healthy adults in a multicenter, randomized, double-blinded, placebo-controlled study. Participants received 3,000 mcg HIV-MAG (gag/pol, env, nef/tat/vif) DNA vaccine coadministered with IL-2 DNA at 0, 250, 000, or 500 mcg (N=22/group) or placebo (N=3/group) intramuscularly by EP at 0, and 3 months boosted by rvsv Gag vaccine or placebo at 6 months. Participants were assessed for reactogenicity, tolerability, and adverse events. CD4+ and CD8+ T-cell responses to HIV potential T-cell epitope (PTE) peptides were measured by intracellular cytokine staining (ICS) 2 weeks after 3rd and 4th vaccinations. Results: EP was generally well tolerated. Local and systemic reactogenicity symptoms were generally mild to moderate. After the 4th vaccination some subjects experienced moderate to severe systemic symptoms and several experienced transient lymphopenia. After DNA prime, CD4+ T-cell responses to any PTE were detected in 77% of subjects and CD8+ T-cell responses in 40%. Gag-specific CD4+ T-cell response rates after DNA prime increased significantly following rvsv Gag boost, from 5% to 86%. Gag-specific CD8+ T-cell responses also increased significantly from 6% after DNA prime to 26% after rvsv Gag boost. IL-2 DNA 500 mcg increased the magnitude of CD8+ T-cell responses compared to no IL-2 DNA (p=0.02). Conclusions: HIV- DNA vaccination given by EP with IL-2 and boosted with HIV-rVSV is safe and immunogenic. DNA/EP prime dramatically improves T-cell responses after a single rvsv boost compared with rvsv homologous prime-boost (HVTN 090). IL-2 DNA does not increase response rates but increases the magnitude of CD8+ T-cell responses. ORAL ABSTRACT SESSIONS 49

50 Oral Abstract Sessions Oral Abstract Session 05: Vaccine, Viral Latency and Cure ORAL ABSTRACT SESSIONS OA05.03 Elicitation of Immune Responses by a DNA/ MVA Vaccine in ART Treated Patients in a Treatment Interruption Trial Harriet L. Robinson, Melanie Thompson 2, Sonya Heath 3, Stephen J. Brown 4, Bentley Sweeton 2, Kathy Williams 2, Pamela Cunningham 3, Miles Viljanen 4, Rahul Basu, Yongxian Xu 5, Suefen Kwa GeoVax, Inc., Smyrna, GA, United States, 2 AIDS Research Consortium of Atlanta, Atlanta, GA, United States, 3 University of Alabama at Birmingham, Department of Medicine, Birmingham, AL, United States, 4 AIDS Research Alliance, Los Angeles, CA, United States, 5 The Hope Clinic of the Emory Vaccine Center, Decatur, GA, United States Background: GV-TH-0, a Phase open-label trial of GOVX-B, a DNA/MVA prime-boost regimen, in HIV infected patients on ART was undertaken to evaluate safety and vaccine-elicited T cell responses, and to explore viral rebound during analytical treatment interruption (TI). Methods: Patients who began ART within 8 months of seroconversion and had sustained plasma HIV- RNA < 50 c/ml for at least 6 months were enrolled. Patients received a total of 4 inoculations at intervals of 8 weeks. 2 of pga2/js7 DNA (3mg) followed by 2 of MVA/HIV62B (0 8 TCID50). At 8 weeks after the last immunization, plus an efavirenz washout if needed, participants entered a TI phase of 2 weeks, after which ART was reinstituted. T cell responses were scored for IFNg or IL2 by flow cytometry following stimulation with Gag, Env and Pol peptides. Responses were considered positive if 2-fold higher than pre-vaccination. Results: 8 of 9 men completed all vaccinations. For the 8, median age was 37.5 yrs, baseline CD4 count was 69/µL (50-62/µl) and all had HIV- RNA < 50 c/ml. Median viral load prior to ART was 5. log0 c/ml ( log0 c/ml). No serious adverse events occurred. After the st or 2 nd MVA/HIV62B immunization, Gag-specific CD8 T cells were boosted over pre-vaccination levels in 7 out of 8 (P< 0.05) whereas Gag-specific CD4 T cells were boosted in 5 of 8 patients (P=0.2). 6 of 8 patients elicited previously undetectable CD8 responses whereas 5 of 8 elicited previously undetectable CD4 responses to Gag epitopes. Gp20 or gp4-specific antibody responses were boosted in 3 of 8 patient and 2 of 8 patients respectively. Excluding one acute seroconverter, the median reduction in HIV- RNA at weeks 2, 6, and 2 compared to pre- ART levels was -2.2, -.3 and -0.8 log0 c/ml. Conclusions: This trial demonstrates the potential for GOVX-B to boost both T cell and antibody responses in a therapeutic setting. A placebo-controlled trial will be required to further assess the therapeutic benefit of the vaccine. OA05.04 Early Initiation of ART in Acute HIV Infection (Fiebig I to III) Does Not Preclude the Development of HIV-specific Cellular Immune Responses Alexandra Schuetz,2, Yuwadee Phuang-Ngern, Rapee Trichavaroj, Nittaya Phanuphak 3,4, Rungsun Rerknimitr 5, Suchada Sukhumvittaya, Surat Jongrakthaitae, Silvia Ratto- Kim 2,6, James Fletscher 3, Eugene Kroon,3, Nitaya Chomchey 3,4, Robert J. O Connell, Viseth Ngauy, Praphan Phanuphak 3,4,5, Nelson L. Michael 6, Jerome H. Kim 6, Mark S. De Souza,3,4, Jintanat Ananworanich 2,3,6, RV254/SEARCH 00 Study Group Armed Forces Research Institute of Medical Sciences United States Component, Retrovirology, Bangkok, Thailand, 2 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 3 SEARCH, Bangkok, Thailand, 4 The Thai Red Cross AIDS Research Centre, Bangkok, Thailand, 5 Chulalongkorn University, Department of Medicine, Bangkok, Thailand, 6 United States Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Silver Spring, MD, United States Background: High viral load (VL) and rapid loss of CD4 + T cells are a hallmark of acute HIV infection. The association between the subsequent emergence of HIV-specific CD8 + T cells and the decrease in VL support that CD8 + T cells are crucial in the initial control of viral replication. However, little is known about the kinetics of emerging T cell responses during early acute infection (Fiebig [F] I to III) and the effect of early initiation of antiretroviral treatment (ART) on the development of HIVspecific T cells. Methods: 28 patients with acute HIV infection ( FI/II, 7 FIII) were enrolled in the RV254/Search 00 study and immediately received ART. HIV-specific immune responses against Gag and Env peptide pools were determined in PBMC using IFN-γ intracellular cytokine staining prior to ART initiation and 6 and 24 months post-art. Results: At time of diagnosis, no HIV-specific T cell responses were detected in FI/II, while in FIII predominantly Gag-specific CD4 + responses were observed in 2/7 (2%) and CD8 + responses in 4/7 (24%) patients. Median plasma VL in FI/II was 5.5 log 0 copies/ml vs. 6.7 log 0 copies/ml in FIII (p=0.00). At 6 and 24 months post-art all patients had undetectable VL. However, 6 months post-art there was no significant difference observed in the frequency of Gag-specific CD4 + and CD8 + responses between patients treated in FI/II (2/ [8%] and 6/ [55%], respectively), and FIII (4/7 [23%], and 8/7 [47%], respectively). The frequency of Gag-specific CD8 + T cell responses was maintained 24 months post-art, primarily in patients that initiated ART in FIII (FI/II CD4 + : 0/, CD8 + : 4/ [36%]; FIII CD4 + : 5/7 (29%), CD8 + : 9/7 [53%] p>0.05). Conclusions: HIV-specific T cell responses, primarily CD8 + mediated against Gag, were detected at FIII but not FI/II. However, after 6 months of ART, both groups had similar numbers of responders maybe due to viremia during the initial post-art period or ongoing viral replication in privileged sites. 50 HIV Research for Prevention 204 HIV R4P

51 Oral Abstract Session 05: Vaccine, Viral Latency and Cure Tuesday, 28 October OA05.05 Engineered Gag-specific T-cell Receptors Redirect Polyclonal CD8 + T-cells to Clear HIV- -infected CD4 + T-cells from ART-treated Patients Hongbing Yang, Sandrine Buisson 2, Giovanna Bossi 2, Gemma Hancock, Rebecca Ashfield 2, Annelise Vuidepot 2, Tara Mahon 2, Peter Molloy 2, Joanne Oates 2, Zoe Wallace, Namir Hassan 2, Bent K. Jakobsen 2, Lucy Dorrell University of Oxford, Nuffield Department of Medicine, Oxford, United Kingdom, 2 Immunocore Ltd, Abingdon, Oxfordshire, United Kingdom OA05.06 LB Structure of HIV- gp20 VV2 in Complex with Human mab 830A Reveals a 5-Stranded Beta Barrel Conformation and Integrinbinding Site Ruimin Pan, Miroslaw K. Gorny 2, Susan Zolla-Pazner 2,3, Xiang- Peng Kong NYU School of Medicine, Department of Biochemistry and Molecular Pharmacology, New York, NY, United States, 2 NYU School of Medicine, Department of Pathology, New York, NY, United States, 3 Veterans Affairs New York Harbor Healthcare System, New York, NY, United States ORAL ABSTRACT SESSIONS Background: HIV- establishes a stable latent reservoir in resting CD4 + T cells that is not eliminated by ART. New immunotherapeutics are needed to deliver a shock and kill in order to reduce viral reservoirs. We evaluated the antiviral efficacy of engineered high affinity immunemobilising monoclonal T cell receptors (ImmTACs) that redirect CD8 + T cells to kill HIV--infected CD4 + T cells. Methods: HIV- ImmTACs comprised engineered high affinity T cell receptors (TCRs) specific for the HIV- gag epitope, SLYNTVATL (SL9) and its common escape variants, fused to a humanized CD3-specific single chain variable fragment. Their antiviral potency was evaluated in viral inhibition and killing assays with ex vivo CD4 + and CD8 + T cells from HLA-A*020-positive ART-treated patients. Fluorescent-labelled TCRs were also used to quantify target epitope density on infected cells. Results: HIV- ImmTACs significantly enhanced ex vivo CD8 + T cell-mediated inhibition of endogenous HIV- replication at low concentrations (-0 nm) and CD8 + /CD4 + ratios of : and :0 (mean, SD: 72%, 4%, p=0.008; 55%, 5%, p=0.004 respectively) despite low epitope expression on the cell surface (< 50/cell). More potent inhibition was achieved when CD8 + T cells from healthy HLA-A*020 + donors were used (mean, SD: 85%, 8%, p< 0.000). Killing of infected cells was confirmed by lack of viral recrudescence after wash-out of the ImmTAC from the culture and was maximal within 48 hours of ImmTAC exposure. Of note, resting infected CD4 + T cells were eliminated almost as efficiently as activated cells (mean 85% vs. 95%). The potency of the ImmTACs was highly correlated with the level of intracellular HIV- gag expression (r 2 =0.48, p< 0.000). Conclusions: ImmTACs can efficiently redirect polyclonal CD8 + T cells to kill activated and resting HIV--infected CD4 + T cells from HIV- positive patients, at low effector:target ratios and low epitope densities. Our data support further evaluation of ImmTACs as a component of HIV- eradication strategies. Background: The first and second variable regions (VV2) of gp20 play vital roles in the function of HIV- envelope (Env). VV2, which harbors multiple glycans and is highly sequence diverse, is located at the Env apex and stabilizes the trimer. It shields V3 and the co-receptor binding site in the pre-fusion state and exposes them upon CD4 binding. Recent structural data, including that of the scaffolded VV2 in complex with PG9/PG6 and the trimeric SOSIPs, suggested that VV2 forms a Greek key motif with 4 beta strands (named strands A-D). However, regions of VV2 including the integrin-binding site were missing from those structural visualizations, and our understanding of the VV2 structure-function is thus not yet complete. Methods: We have determined a crystal structure of the VV2 scaffold (VV2 ZM09 -FD6) in complex with 830A, an anti-vv2 human mab with an epitope known to overlap the integrin-binding site. Results: Our complex structure revealed that VV2 has a 5-stranded beta barrel structure with the region of the integrin-binding site forming a kink (AAs 78-80) followed by an additional beta strand (strand C ) before strand D. The complete barrel structure naturally presents the glycans on its outer surface and packs conserved hydrophobic residues, including the RV44 sieve residue Ile 8, in its core,. The two lengthvarying regions in V and V2 form two extended loops towards one end of the barrel. The epitope of 830A is discontinuous with three segments: it centers at Thr 75, Tyr 77, Leu 79 and Asp 80 at the kink overlapping the integrin-binding site (AAs 79-8) and also includes residues Arg 53 and Val 54 in V and Leu 93, Ile 94 and Ser 95 at the C-terminus of V2. Conclusions: The VV2 region forms a 5-stranded beta barrel, a unique structure that can function as an independent module. 5

52 Oral Abstract Sessions Oral Abstract Session 06: B Cell Repertoires for Protection ORAL ABSTRACT SESSIONS OA06.0 Evolution of Antigen-specific B-cell Receptor Repertoires in Early SIV Infection Eva J. Archer,2,3, Rachael Bashford-Rogers 2, Brenna Hill, Daniel C. Douek, Paul Kellam 2, Richard A. Koup Vaccine Research Center, NIAID, NIH, Bethesda, MD, United States, 2 Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 3 University of Cambridge, Cambridge, United Kingdom Background: Broadly neutralizing antibodies to HIV arise only in a fraction of cases after years of infection, while early antibody responses to HIV infection typically do not neutralize circulating virus or provide protection from infection. Detailed understanding of the early antibody response can inform vaccine and immunogen design. We use nextgeneration sequencing to track the evolution of the antibody repertoire generated in response to SIV infection in macaques. Methods: Naive, memory, and antigen-specific B cells were sorted from blood, lymph nodes, and bone marrow of rhesus macaques infected with SIVmac25 at 4, 0, and 24 weeks post infection. B cell receptor heavy chain sequences were amplified using Ig-specific primers and sequenced using MiSeq 2x300bp paired end reads to obtain full length VDJ sequences. Sequences were analysed using a bioinformatics platform to track V, D, and J gene usage; mutation from germline; CDR3 sequence and length; and clonality and diversity of the sample populations over time. Results: Antigen-specific B cells were first detected in the bone marrow, blood, and lymph nodes at 4 weeks post infection using a SIV gp40 trimeric probe. The fraction of SIVgp40 positive memory B cells increased throughout infection, up to.5% of total B cells in lymph nodes at 24 weeks post infection. Antigen specific B cells show accumulation of mutations in the V H gene compared to nonspecific memory B cells, and organize naturally into clusters of clonally related sequences from acute and early chronic infection. Conclusions: This study is a high-resolution analysis of memory and antigen-specific B cells in different spatial compartments showing the dynamics of the humoral immune response from acute to early chronic SIV infection. OA06.02 LB Role of Intestinal Microbiota in Shaping the B Cell Repertoire in HIV Infection and Env Vaccination Larry (Huaxin) Liao, A.M. Trama, W.B. Williams, M.A. Moody, Nathan Vandergrift, G.D. Tomaras, D.J. Marshall, T. Gurley, J. Whitesides, J. Eudailey, A. Foulger, R. Parks, C. Stolarchuk, K.E. Lloyd, K. Soderberg, J.R. Mascola 2, R. Koup 2, L. Corey 3, G.B. Nabel 2, P. Gilber 4, C. Morgan 3, J. Maenza 3, M. Keefer 5, S. Hammer 6, G. Churchyard 7, D.C. Montefior, B.S Graham 2, L.R. Baden 8, T.B. Kepler 9, B.F. Haynes Duke University Medical Center, Human Vaccine Institute, Durham, NC, United States, 2 National Institute of Allergy and Infectious Diseases, Vaccine Research Center, Bethesda, MD, United States, 3 University of Washington, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 University of Washington, SCHARP, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 5 University of Rochester Medical Center, Division of Infectious Disease, Rochester, NY, United States, 6 Columbia University Medical Center, New York, NY, United States, 7 The Aurum Institute, Johannesburg, South Africa, 8 Brigham and Women s Hospital, Boston, MA, United States, 9 Boston University, Boston, MA, United States Background: The memory B cells in intestine contain a subset of cells reactive with commensal bacteria. In acute HIV- infection (AHI), virus replication is prominent in the gastrointestinal tract with early depletion of CD4+ T cells and destruction of germinal centers. Antibodies in serum and mucosal fluid in AHI are targeted to HIV- Env gp4, are cross-reactive, non-neutralizing and do not select viral escape. Thus we hypothesize that pre-transmission commensal bacteria antibodies that cross-react with HIV- antigens may shape the B cell response to HIV-. Methods: Variable region gene segments of Ig heavy- and light-chain (V H and V L ) were isolated from single plasma cells and memory B cells sorted from individuals with AHI and early HIV- infection, and from vaccinees who received the prime-boost vaccination in HVTN Phase II trials with VRC DNA and rad5 vaccine containing clades A, B and C env gp40 genes. The Isolated V H and V L genes were expressed as recombinant mabs for characterization. Results: The predominant Env antibody response in blood in early HIV- infection is to gp4. Of 42 mabs isolated from terminal ileum plasma cells, 9 (5%) were HIV--reactive, 4 of which were gp4-reactive, and of these, (79%) were cross-reactive with gut flora. One gut flora antigen that reacted with gp4 antibodies was identified as E.coli RNA polymerase. From the vaccinees with VRC DNA prime and rad5 boost (HVTN204 and HVTN082), 90.2% of 254 antibodies derived from HIV- Env-specific memory B cell were gp4-reactive. Vaccine-induced gp4 mabs failed to neutralize HIV- transmitted/founder viruses, and remarkably, 82% of the gp4 antibodies were cross-reactive with intestinal commensal -bacterial antigens. Conclusions: Acute HIV- infection and Env gp40 vaccination may have induced dominant HIV- gp4 antibody responses resulted in part from expansion of preexisting cross-reactive, mutated, memory B clones that were triggered pre-hiv infection by commensal bacterial antigens. 52 HIV Research for Prevention 204 HIV R4P

53 Oral Abstract Session 06: B Cell Repertoires for Protection Tuesday, 28 October OA06.03 Plasmablast Phenotype and Mucosal Antibodies to V2 in Vaccine-induced Protection Against SIVmac25 Luca Schifanella,2, Nicolo` Binello, Monica Vaccari, Shari N. Gordon, Francesca Caccuri, Matthew Blackburn, Melvin Doster, Namal Liynage, Poonam Pegu, Xiaoying Shen 3, Georgia D. Tomaras 3, David Venzon 4, Don Stablien 5, Susan W. Barnett 6, Dan Barouch 7, Sanjay Phogat 8, Genoveffa Franchini NIH/NCI, Vaccine Branch, Bethesda, MD, United States, 2 Università degli Studi di Milano, Department of Biomedical and Clinical Sciences - Section of Infectious Diseases and Immunopathology L. Sacco, Milano, Italy, 3 Duke Human Vaccine Institute, Durham, NC, United States, 4 NIH/NCI, Biostatistics and Data Management Section, Bethesda, MD, United States, 5 The EMMES Corporation, Rockville, MD, United States, 6 Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United States, 7 Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA, United States, 8 Sanofi Pasteur, Swiftwater, PA, United States Background: We have recently recapitulated the RV44 vaccine efficacy in a SIV mac25 model. In our study, rectal anti-cyclic V2 IgG antibodies correlated with a decrease risk of SIV mac25 acquisition (p=0.0063). Analysis of the homing markers on plasmablast (PB) resulted in a higher frequency of alpha 4 beta 7 + PBs in animals with higher levels of IgG and IgA to cyclic V2 in rectal mucosal. Methods: We investigated the homing potential in 5 different vaccine strategies by measuring alpha 4 beta 7 and CXCR3 as markers for gut mucosa and inflammatory sites, respectively in a total of 8 macaques. The immunoglobulin (Ig) expression on PB and the mucosal antibody responses were assessed in all groups. We studied a cohort of macaques immunized 4 times with ALVAC-SIV and twice with gp20/alum or gp20/mf59. A second cohort was immunized twice with DNA-SIV/ gp20/alum or once Ad26-SIV, and boosted with ALVAC-SIV/gp20/ alum twice. Finally, an additional group was immunized 4 times with NYVAC-SIV boosted twice with gp20/alum. Results: Both ALVAC-SIV/gp20/alum and DNA-SIV/gp20/alum immunized animals were protected from SIV mac25 mucosal acquisition (p=0.029 and p=0.04, respectively). However, no protection was observed with ALVAC-SIV/gp20/MF59, NYVAC-SIV/gp20/alum and Ad26 primed regimens. DNA-SIV/gp20/alum and ALVAC-SIV/gp20/ alum immunizations increased the frequency of alpha 4 beta 7 plasmablasts (p=< and p=0.05) while ALVAC-SIV/gp20/MF59, NYVAC- SIV/gp20/alum increased the frequency of CXCR3+ plasmablasts (p=0.000 and p=0.004 respectively). We are completing Ad26-SIV/ gp20 and DNA-SIV/gp20/alum studies as well as the analysis of Ig expression and correlations with mucosal antibody responses. Conclusions: Preliminary results suggest that different vaccine modalities are able to alter the plasmablast homing and the effector functions of the antibody response at mucosal sites that may correlate with protection. OA06.04 Use of Enzyme-digested Virus-like Particles as Probes for Flow Cytometric Sorting of HIVspecific Neutralizing Ab-producing B-cells Evan M. Cale, Nicole A. Doria-Rose, Tommy Tong 2,3, Ema T. Crooks 2,3, Richard Nguyen, David R. Ambrozak, Stephen P. Perfetto, Mario Roederer, James M. Binley 2,3, John R. Mascola NIAID, National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States, 2 Torrey Pines Institute for Molecular Studies, San Diego, CA, United States, 3 San Diego Biomedical Research Institute, San Diego, CA, United States Background: Isolating new broadly neutralizing antibodies (bnabs) provides fresh insights for rational HIV- vaccine design. Several sites of vulnerability to bnabs are now well defined, and it is likely that additional targets exist. Virus-like particles (VLPs) that express Env trimers display all of the natural epitopes and are therefore attractive candidates for use as probes. However, VLP surfaces are also decorated with nonfunctional forms of HIV Env. Exposing VLPs to proteases clears nonfunctional Env, leaving native trimeric Env intact. The resulting trimer-vlps preferentially bind to nabs compared to non-neutralizing antibodies and therefore may have gained sufficient selective power for use as B cell probes to identify new bnabs. Methods: PBMC from HIV-infected patients were stained with the enzyme-digested trimer-vlps, and VLP-positive memory B cells were sorted by flow cytometry. Multiplex RT-PCR was performed on cell lysates to amplify kappa, lambda, and heavy chain genes. Cloned antibodies were assessed for neutralization breadth and potency against a panel of Env-pseudoviruses using the TZM-bl neutralization assay. Results: VLPs were used to sort B cells from the donor of the broad CD4bs antibody VRC3. The sort yielded relatives of VRC3, as well as unrelated sequences that exhibited high sequence divergence from germline VH genes and/or long CDR3 regions, which are characteristics of known bnabs. Cloning and characterization of these Abs is ongoing. Conclusions: Unlike many previously used B cell probes, VLPs express multiple epitopes including those for trimer specific antibodies which allows for simultaneous sorting of B-cells of different specificities. Additionally, these probes are devoid of non-functional forms of Env and therefore preferentially bind to neutralizing Ab-producing B cells. VLPs therefore hold great promise for isolating novel HIV-specific bnabs that can better inform vaccine design. ORAL ABSTRACT SESSIONS 53

54 Oral Abstract Sessions Oral Abstract Session 06: B Cell Repertoires for Protection ORAL ABSTRACT SESSIONS OA06.05 Viral Escape Pathways from Broadly Neutralising Antibodies Targeting the HIV Envelope Cleavage Site Enhance MPER Mediated Neutralisation Constantinos Kurt Wibmer,2, Daniel J. Sheward 3, Jinal N. Bhiman,2, Nonkululeko Ndabambi 3, Debra H. Elliot 4, Julie Rouelle 4, Ashley Smira 4, Salim S. Abdool Karim 5, James E. Robinson 4, Lynn Morris,2,5, Carolyn Williamson 3,5, Penny L. Moore,2,5 Centre for HIV & STIs, National Institute for Communicable Diseases, NHLS, Johannesburg, South Africa, 2 Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 3 Institute of Infectious Disease and Molecular Medicine (IIDMM) and Division of Medical Virology, University of Cape Town and NHLS, Cape Town, South Africa, 4 Tulane University Medical Center, Department of Pediatrics, New Orleans, LA, United States, 5 Centre for the AIDS Programme of Research in South Africa (CAPRISA), Nelson R Mandela School of Medicine, University of KwaZulu-Natal, Durban, South Africa Background: A preventative HIV- vaccine will likely elicit broadly neutralising antibodies (bnabs). The diverse modes of neutralisation by bnabs leads to variable escape pathways in each epitope. Documenting escape and identifying common elements for each site of vulnerability will inform immunogen design. Here, we map a newly described bnab specificity at the gp20-gp4 interface, and define viral escape in CAPRISA donor CAP248. Methods: Escape mutations were identified from longitudinal singlegenome envelope sequences, and confirmed by mutagenesis and neutralisation assays. CAP248 plasma responses were mapped longitudinally to determine the kinetics of bnab maturation in response to neutralisation escape. A monoclonal antibody (mab), CAP B, was isolated by B-cell culture and characterised. Results: The CAP B mab recapitulated the plasma breadth, but was significantly less potent due to an unusual neutralisation plateau. The CAP B mab was sensitive to deletion of the N6 glycan in most heterologous viruses, but this mutation had a less significant effect on CAP248 plasma, suggesting a swarm of cleavage site directed bnabs in CAP248 with differential glycan dependence. Viral escape mutations from CAP248 bnabs accumulated in the gp60 cleavage sites (positions 500, 502, 505, and ) as well as proximal sites in gp4. These changes collectively resulted in 00-fold enhanced neutralisation by the MPER bnabs 4E0 and 0E8. The binding of 4E0 or 0E8 to captured virus enhanced CAP B binding. Conclusions: New bnabs targeting the gp20-gp4 interface, such as those in CAP248, have provided an additional target for HIV- vaccines. The enhancement of MPER mediated neutralisation by CAP248 escape mutations in the cleavage site suggests a synergistic relationship between cleavage site and MPER bnabs, similar to that between VV2 and CD4 binding site bnabs. Delineation of CAP248 virus-antibody coevolution may thus provide a blueprint for the co-induction of these two antibody classes. OA06.06 Maturation Pathways for the Broad and Potent Antibody 0E8 using a Combination of Next Generation Sequencing, Bioinformatics and Functional Analysis Cinque S. Soto, Gilad Ofek, M. Gordon Joyce, Baoshan Zhang, Krisha McKee, John R. Mascola, Peter D. Kwong NIAID, NIH, Vaccine Research Center, Bethesda, MD, United States Background: Antibodies that target the membrane-proximal external region (MPER) of the HIV- gp4 subunit represent one of the few categories of antibodies that effectively neutralize HIV-. Such antibodies provide potential templates to guide vaccine strategies, but details of their generation and maturation have been unclear. Methods: From the PBMCs of a single time point of donor N52, we used 454 Pyrosequencing to obtain the sequences of the variable domains of heavy and light antibodies from B cell transcripts. Bioinformatics sieving of gene families and structure-based sequence signatures delineated potential transcripts from the maturation pathway of the effective MPERdirected antibody 0E8. We calculated maturation intermediates along these pathways, synthesized heavy and light chains, and used transient transfection to express antibodies, which we assessed for neutralization and for MPER-peptide affinity. Results: We identified 235 heavy chain sequences with V, D and J segments matching the mature 0E8 antibody and 47,69 light chain sequences with V and J segments matching the mature 0E8 antibody. These sequences were further sieved with structural signatures. Pathways - based on maximal intermediates, minimal sequence reversions, and minimal N-nucleotide additions - for heavy and light chain were identified, and we paired heavy and light chain sequences for the unmutated common ancestor (UCA) along with three inferred heavy and light chain sequences. Unexpectedly, the CDR H3 of the UCA differed substantially from the mature sequence. Nonetheless, all of the paired intermediates, including the UCA, showed binding to MPER peptide, while only the two most mature intermediates showed neutralization. Binding to MPER in the context of lipid micelles, meanwhile, mirrored that of neutralization. Conclusions: The combination of cross-sectional NGS data, bioinformatics, and functional analysis can be used to define the maturation pathways for an antibody, even with cross-sectional data. 54 HIV Research for Prevention 204 HIV R4P

55 Oral Abstract Session 07: Risk and Prevention for MSM Tuesday, 28 October OA07.0 Characteristics of Young Black Men who Have Sex with Men Enrolled in HPTN 06 Christopher Chauncey Watson, Jonathan Paul Lucas 2, Darrell P. Wheeler 3 The George Washington University, Public Health Research Clinic, Washington, DC, United States, 2 FHI 360, Science Facilitation, Durham, NC, United States, 3 Loyola University Chicago, School of Social Work, Chicago, IL, United States Background: Men who have sex with Men (MSM) comprise the largest proportion of new HIV diagnosis in the United States (CDC, 202). The prevalence among Black MSM is higher than that of Latino or White MSM (Jeffries, 202). Most studies documenting HIV incidence among Black MSM have been based on cross-sectional studies of HIV surveillance. Very few studies have estimated incidence among young Black MSM, who continue to be disproportionately infected at higher rates than other racial/ethnic groups. Methods: HPTN 06 (The BROTHERS Study) was a six US city study designed to determine the feasibility and acceptability of a multicomponent HIV prevention intervention for Black MSM. Men were recruited from 7/09 to 0/0 through a multi-pronged approached. Once found eligible, men participated in 3 visits (baseline, 6 mo, 2 mo) visits which included ACASI interviews, demographic collections, HIV/ STI testing and counseling, social/sexual network questionnaire and the opportunity to work with a Peer Health Navigator. Results: Of the 553 Men recruited in HPTN 06, 59 were < 30. Overall HIV incidence for HPTN 06 was 3% but was 5.9% among < 30 y.o. Younger men had more STIs, no usual place for healthcare and more identified barriers to receiving health care. A multivariate analyses found HIV incidence was associated with younger age (HR 3.4, CI:.4-8.3) and unprotected receptive anal intercourse with HIV positive partners or unknown status partner (HR 4.,.9-9.). Over 50% of the individuals reported no work and no health coverage. Conclusions: In the largest longitudinal cohort of Black MSM to date in the US, HIV incidence was highest among Young BMSM. We found significant drivers for HIV acquisition among Young Black MSM to include; access to health care services, higher sexual risk behaviors, and occurrence of STIs. Additional research is needed to focus primarily on this population to develop effective prevention strategies. OA07.02 Through the Rabbit Hole: Considering the Situational Experience of Risk among Men who Have Sex with Men in the Context of HIV Prevention Martina Brostrom, Nicola Desmond 2,3 UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva, Switzerland, 2 Malawi-Liverpool-Wellcome Trust, Blantyre, Malawi, 3 Liverpool School of Tropical Medicine, Department of Clinical Sciences, Liverpool, United Kingdom Background: After 3 decades of research and programming for MSM, in 204, HIV remains a major public health challenge in this population. Even in countries where MSM have access to HIV services, civil liberties and community structures, prevalence rates are consistently high. This warrants development and scale up of bespoke, innovative and meaningful HIV combination prevention programmes that better respond to the diverse social and health needs of MSM. A better insight into the experiences of risk and the impact on sexual practices among MSM is required to inform such approaches. This paper provides a metasynthesis of the linkages between the situational experience of risk for MSM and unprotected sexual intercourse and explores the implications for HIV prevention. Methods: A systematic search of nine electronic databases was conducted and complemented by a free text search on Google scholar on the name and publications of key authors and an inspection of the references of identified articles. Six qualitative studies that investigated the situational experience of risk among MSM were identified. A metaethnographic synthesis was undertaken to describe, analyse and interpret the studies. Results: The synthesis resulted in the identification of 6 themes: losing control; relationship building; fatalism, homophobia and stigma of HIV; prevention literacy and confusion; risk management strategies and the biography of individuals. Conclusions: MSM are well aware of the risks of contracting HIV through unprotected anal intercourse and deploy prevention strategies to stay safe and healthy, but what they conceive as safe and risk is different from that of medical professionals. Frequently MSM are ambiguous about risk and this is reflected in their sexual practices. Those working in HIV prevention must be aware of the lay risk management strategies deployed by MSM and engage with them. ORAL ABSTRACT SESSIONS 55

56 Oral Abstract Sessions Oral Abstract Session 07: Risk and Prevention for MSM ORAL ABSTRACT SESSIONS OA07.03 Sexual Behaviour Profile of Gay and Other Men who Have Sex with Men Enrolled in the PROUD Pre-exposure Prophylaxis Open-label Pilot Study in England Mitzy Gafos, David Dolling 2, Monica Desai 2, Gemma Wood 2, David Dunn 2, John Saunders 3, Nicola E. Mackie 4, Alexandra Meijer 5, Amanda Clarke 6, Christine Bowman 7, Charles Lacey 8, Lisa Southon 9, Clare Oakland 0, Chris Higgs, David White 2, Iain Reeves 3, Michael Brady 4, Julie Fox 5, Anthony Nardone 6, on behalf of the PROUD Study Team MRC Clinical Trials Unit at UCL, Institute of Clinical Trials & Methodology, London, United Kingdom, 2 MRC Clinical Trials Unit at UCL, London, United Kingdom, 3 Ambrose King Centre/Barts Sexual Health Centre, London, United Kingdom, 4 Jefferiss Wing, St Mary s Hospital, London, United Kingdom, 5 56 Dean Street Clinic, London, United Kingdom, 6 Elton John Centre, Brighton & Sussex University Hospital Trust Sussex House, Brighton, United Kingdom, 7 Royal Hallamshire Hospital, Dept of GU Medicine, Sheffield, United Kingdom, 8 Monkgate Health Centre, Dept of GU Medicine, York, United Kingdom, 9 Manchester Centre for Sexual Health, Manchester, United Kingdom, 0 Centre for Sexual Health & HIV Research, Mortimer Market Centre, London, United Kingdom, John Hunter Clinic, St. Stephen s Centre, Chelsea & Westminster Hospital, London, United Kingdom, 2 Medical Innovation Development Research Unit (MIDRU), Birmingham Heartlands Hospital, Birmingham, United Kingdom, 3 Homerton Sexual Health Services, London, United Kingdom, 4 Caldecot Centre, London, United Kingdom, 5 St Thomas Hospital, London, United Kingdom, 6 Public Health England, London, United Kingdom Background: The public health benefit of PrEP will depend on effective targeting of higher risk individuals. We report the baseline sexual behaviour of men enrolled in England s PROUD pilot study. Methods: In PROUD, eligible HIV negative gay/msm, aged 8+, who report recent and intended condomless anal sex, are randomized to receive Truvada as PrEP immediately or after 2 months. Results: The PROUD cohort of 470 men are a median age of 35 (IQR 34-36), 79% white, 58% UK born, 60% university educated, 82% employed, 47% in a relationship and 96% self-identify as gay. Anal sex was reported with a median of 0 (IQR 5-20) partners total, and 7 (IQR 2-5) new partners in the last 90 days. Condoms were not used with a median of 2 (IQR -5) partners for receptive sex and 3 (IQR -6) for insertive. Reasons for no condom at last condomless sex were: 66% less pleasure, 5% don t like condoms, 33% partner didn t like condoms, 27% condoms not discussed, 23% and 22% under influence of drugs or alcohol (68% consumed alcohol weekly and 73% used recreational drugs in the last 90 days). At last condomless sex, 45% assumed partner to be HIV-, 27% HIV+ on ART, 23% HIV unknown, 5% HIV+ not on/unknown ART. When having condomless anal sex, 29% felt at little or no risk, 47% somewhat at risk, 23% at high risk. Risk reduction strategies reported were: 39% use condoms, 26% ask partner to use condom, 38% chose HIV- partners, 29% use strategic positioning, 23% seek partners on ART; only 6% reported no strategy. In the last year 39% had used PEP (9% >=2 times); 87% had visited a clinic >=2 times for HIV and 82% for STI testing. 37% reported a bacterial rectal STI, which was associated with higher numbers of condomless sex partners (OR.09 receptive p< 0.00,.04 insertive p=0.0). Conclusions: PROUD is enrolling gay men who are selective condom users with high rates of STIs, who despite regular clinic attendance and use of risk reduction strategies, including sero-sorting and PEP, are likely to be at high risk of HIV acquisition. OA07.04 Structural Stigma Affects Access to Pre- and Post-exposure Prophylaxis and HIV Risk among Men who Have Sex with Men (MSM) in the United States Catherine Oldenburg, Amaya Perez-Brumer 2, Mark Hatzenbuehler 2, Douglas Krakower 3, David Novak 4, Matthew Mimiaga,5,6, Kenneth Mayer 3,5,7 Harvard School of Public Health, Department of Epidemiology, Boston, MA, United States, 2 Columbia University Mailman School of Public Health, Department of Sociomedical Sciences, New York, NY, United States, 3 Beth Israel Deaconess Medical Center, Boston, MA, United States, 4 OLB Research Institute, Cambridge, MA, United States, 5 The Fenway Institute, Boston, MA, United States, 6 Massachusetts General Hospital, Department of Psychiatry, Boston, MA, United States, 7 Harvard School of Public Health, Department of Global Health and Population, Boston, MA, United States Background: Stigmatizing social environments (structural stigma) are hypothesized to affect HIV risk through several pathways, including decreased access to HIV prevention services. Methods: In August 203, members of the largest MSM social networking site in the US completed a survey about current HIV prevention practices. State-level structural stigma was based on a previously validated composite index: ) density of same-sex couples; 2) proportion of Gay-Straight alliances per public high school; 3) 4 state laws related to sexual orientation; and 4) public opinion toward homosexuality. This information was linked to survey responses via participants state of residence. Multivariable logistic generalized estimating equations were used to assess the relationship between structural stigma and 3 outcomes: () unprotected anal sex (UAS) in the previous 3 months; (2) having taken or heard of HIV pre-exposure prophylaxis (PrEP) and postexposure prophylaxis (PEP); and (3) comfort discussing male-male sexual behavior and HIV prevention with primary care providers, adjusting for demographic and socioeconomic covariates. Results: Among 5,32 HIV-uninfected MSM, lower levels of structural stigma were associated with decreased odds of UAS (aor 0.97, 95% CI ), greater odds of awareness of (aor.05, 95%CI ) and taking PEP (aor.0, 95% CI.0 to.9), awareness of (aor.04, 95% CI.0-.08) and taking PrEP (aor.3, 95% CI ), and comfort discussing male-male sex (aor.08, 95% CI.05-.) and HIV prevention strategies (aor.05, 95% CI ) with primary care providers. Conclusions: MSM living in highly stigmatizing environments report greater individual-level risk behaviors and decreased knowledge and access to biomedical HIV prevention strategies, placing them at heightened vulnerability for HIV. Legal reforms to protect sexual minorities are needed to reduce HIV risk and support existing HIV prevention efforts among MSM. 56 HIV Research for Prevention 204 HIV R4P

57 Oral Abstract Session 07: Risk and Prevention for MSM Tuesday, 28 October OA07.05 Multiple HIV Counselling Sessions and Safe Sex Practice Helped Bangkok Men who Have Sex with Men Stay HIV-uninfected: Thailand Wipas Wimonsate, Sarika Pattanasin, Anuwat Sriporn, Pikunchai Luechai, Kesinee Satumay, Narongritt Tippanonth, Nutthawoot Promda, Anchalee Varangrat, Anupong Chitwarakorn 2, Timothy H. Holtz,3 Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Ministry of Public Health, Department of Diseases Control, Nonthaburi, Thailand, 3 Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and Prevention, Atlanta, GA, United States Background: In prior surveys, Thai men who have sex with men (MSM) have had a low level of HIV awareness and prior history of testing, and significant levels of unsafe sex. We hypothesized that reinforcement of HIV risk-reduction strategies through multiple counselling sessions would be associated with remaining HIV uninfected. Methods: We enrolled Thai MSM from the Bangkok metropolitan area into a five-year cohort study with four-monthly visits (maximum of 6 visits). At every visit, MSM received a comprehensive counselling session: HIV transmission information; risk-reduction strategies; HIV testing; and provision of condoms and lubricants. Logistic regression was used to investigate if number of visits was associated with remaining HIV uninfected. Results: From April 5, 2006 to March 24, 204,,260 HIV-uninfected MSM, age 8 years, were enrolled, and followed and tested for HIV every four months. The mean number of follow-up visits during the study period was (Standard Deviation, SD, 5). We detected 239 MSM with incident HIV infection. Logistic regression showed the odds of remaining HIV uninfected were.3 times as high for each subsequent visit (Adjusted Odds Ratio, AOR,.29, 95% Confidence Interval, CI, ), and associated with pre-study practice of only insertive sex (AOR 2.53, 95% CI ), protected anal sex (AOR.75, 95% CI ), and never joining in group sex (AOR.73, 95% CI ), when adjusted for pre-study age group, education level, employment, living arrangement, recreational drug use, coerced sex, history of STI testing, history of HIV testing, and HIV knowledge. Conclusions: Regardless of other pre-study demographic and behavioral factors, more visits and pre-study safe sex practices were associated with a higher odds of remaining HIV uninfected. Safe sex messages and retention in comprehensive HIV testing services should be strengthened among Thai MSM. OA07.06 LB Project PrEPARE: High Levels of Medication Adherence with Continued Condomless Sex in U.S. Men who Have Sex with Men in an Oral PrEP Adherence Trial Kenneth H. Mayer,2,3, Steven Safren,4,5, Jessica Haberer 3,6, Steven Elsesser, William Clarke 7, Craig W. Hendrix 7, Mark Marzinke 7, Matthew J. Mimiaga,4,8 Fenway Health, The Fenway Institute, Boston, MA, United States, 2 Beth Israel Deaconess Medical Center, Division of Infectious Diseases, Boston, MA, United States, 3 Harvard Medical School, Medicine, Boston, MA, United States, 4 Massachusetts General Hospital, Psychiatry, Boston, MA, United States, 5 Harvard Medical School, Psychiatry, Boston, MA, United States, 6 Massachusetts General Hospital, Medicine, Boston, MA, United States, 7 Johns Hopkins University School of Medicine, Pathology, Baltimore, MD, United States, 8 Harvard School of Public Health, Epidemiology, Boston, MA, United States Background: Pre-exposure prophylaxis (PrEP) has been shown to decrease HIV incidence in MSM, but optimal protection is dependent on medication adherence. The data presented here are from a pilot study, designed to assess whether an intervention based on Life-Steps, an effective intervention to enhance HAART adherence in HIV-infected patients, could improve PrEP adherence among men who have sex with men (MSM). who reported recent condomless anal sex. Methods: Between /202 and 2/203, 55 Boston-area HIV- MSM who were interested in initiating PrEP were enrolled in Project PrEPARE, and randomized to Life-Steps which included 4 weekly sessions that addressed barriers and facilitators of PrEP adherence, or to a control condition that provided supportive counseling. Adherence was monitored by Wisepill, and unprotected sex was assessed via daily text messaging. Results: Participants were primarily White (94%) and well-educated (64% had completed college), and sociodemographic factors were similar in both randomized groups. Over the 6 months observation, 90% of the MSM in each group took at least 80% of their daily pills on a weekly basis, as measured by Wisepill and drug levels, and this did not differ by study condition. Condomless sex rates did not change significantly in either group over time. About 90% of the time during the study, the men in both groups either had drug levels consistent with taking most of their PrEP doses and/or used condoms during sex. No incident HIV infections occurred during the study. Conclusions: Most MSM in this study demonstrated levels of PrEP adherence consistent with protective benefit, whether they received Life-Steps or supportive counseling. Given the high rates of adherence in both groups, the data suggest that MSM who elect to receive open-label PrEP can be highly adherent and self-protective. New PreP adherence interventions should focus on at risk persons who anticipate or demonstrate difficulties with routine medication adherence. ORAL ABSTRACT SESSIONS 57

58 Oral Abstract Sessions Oral Abstract Session 08: Correlates of Protection and Exposure ORAL ABSTRACT SESSIONS OA08.0 Evaluation of Mucosal Tissue Explants as ex vivo Surrogates of in vivo Vaccination of Nonhuman Primates (NHPs) and Humans Carolina Herrera, Ronald Veazey 2, Alexandra Schuetz 3, Natalia Olejniczak, Agnès-Laurence Chenine 4, Sorachai Nitayaphan 3, Jaranit Kaewkungwal 5, Punnee Pitisuttithum 5, Supachai Rerks- Ngarm 6, Robert J. O Connell 4, Jean-Louis Excler 4, Jerome H. Kim 4, Robin Shattock Imperial College, Infectious Diseases, London, United Kingdom, 2 Tulane National Primate Research Center, Tulane, LA, United States, 3 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 4 Walter Reed Army Institute of Research, Rockville, MD, United States, 5 Mahidol University, Bangkok, Thailand, 6 Ministry of Public Health, Bangkok, Thailand OA08.02 Evolutionary Analysis Identifies an MX2 Haplotype Associated with Natural Resistance to HIV- Infection Irma Saulle, Mara Biasin, Federica Gnudi, Salomè Ibba, Micaela Garziano, Manuela Sironi 2, Daria Trabattoni, Sergio Lo Caputo 3, Francesco Mazzotta 3, Antonio Caruz 4, Luca De Gioia 5, Mario Salvatore Clerici 6 University of Milan, Biomedical and Clinical Sciences, Milan, Italy, 2 Scientific Institute IRCCS MEDEA, Bosisio Parini, Milan, Jamaica, 3 S. Maria Annunziata Hospital, Florence, Italy, 4 University of Jaen, Immunogenetic Unit, Jaen, Spain, 5 University of Milan-Bicocca, Biotechnology and Biosciences, Milan, Italy, 6 Don Gnocchi Foundation, Milan, Italy Background: HIV vaccine trials have revealed the need to establish better correlations between NHP and human studies. This project aimed to use ex vivo cervicovaginal and colorectal explants from NHPs and from participants of the ongoing trial RV305 to assess B-cell responses elicited upon vaccination with ALVAC/AIDSVAX B/E. Having shown that in vivo vaccination elicits specific anti-gp20 HIV-CM244 IgG and IgA systemically and mucosally, we assessed the ex vivo infectibility of NHP and human explants and the neutralization profile of tissue culture supernatants Methods: Six Rhesus macaques were immunized with ALVAC/AIDSVAX B/E following RV44 regime. Colorectal and cervicovaginal biopsies were obtained 4 weeks pre-vaccination, and 2 weeks after each immunization. Serum was collected at the same time points. Vaginal and colorectal biopsies were obtained at week 26 from participants enrolled in RV305 having been vaccinated at weeks 0 and 24 with ALVAC/AIDSVAX B/E, AIDSVAX B/E, or ALVAC, or placebo in each group. Neutralization profile of non-infected explant supernatants against two CRF0_AE isolates was assessed in TZM-bl cells. Explants were challenged ex vivo with SHIVBaL or SHIVSF62P3 or with HIV- CM235-LucRT2A and cultured for 5 days. Infectibility was determined by measurement of p27/p24 viral antigen in culture supernatants. Limited unblinding of RV305 samples was performed allowing correlation of samples to groups Results: Moderate neutralization was observed in all NHP and human explants and in NHP serum. Furthermore, limited ex vivo infectibility was detected in NHP mucosal explants after vaccination. Variable levels of infection were observed in human samples corresponding potentially to individuals having received vaccine or placebo within each group Conclusions: While awaiting for further unblinding, this study supports our hypothesis that ex-vivo mucosal tissue models could represent a new tool to assess mucosal B-cell responses to vaccination in NHP studies and during clinical trials Background: The human and macaque MX2 (myxovirus resistance 2) proteins efficiently restrict HIV- and other simian immunodeficiency viruses, but have a modest effect against retroviruses that infect nonprimate species. This observation points to species-specific virus-host interactions that may result from evolutionary arms races. This research project, thus, aimed to demonstrate the relevance of MX2 variants to HIV- infection susceptibility Methods: We analysed: ) evolutionary history of MX2 in placental mammals (PAML analysis); 2) structural models of MX2; 3) genotyping analysis of rs in the MX2 gene in 3 independent case-control cohorts of HIV- exposed seronegative individuals (HESN); and 4) in vitro HIV- replication plus IFNα-stimulated MX2 mrna expression in peripheral blood mononuclear cells (PBMC) from healthy controls (HC) grouped according to their MX2 genotype. Results: MX2 evolved adaptively in mammals with distinct sites representing selection targets in rodents and primates; pervasive selection mainly involves residues in loop 4, previously shown to carry antiviral determinants. We also demonstrated that recent distinct selective events have driven the frequency increase of two haplotypes in human populations of Asian and European ancestry. The Asian haplotype carries a susceptibility allele for melanoma; the European haplotype is tagged by rs , an intronic variant. By analysis of three independent HESN cohorts with different geographic origin and exposure route we verified that the ancestral (G) allele of rs protects from HIV- infection with a recessive effect (combined p value of.55x0-4 ). In line with these findings, the G allele is associated with lower in vitro HIV- replication and increased MX2 expression. Conclusions: Results herein establish a role for MX2 as a central element of antiviral response in mammalian species and a possible target for therapeutic intervention in HIV- treatment and prevention 58 HIV Research for Prevention 204 HIV R4P

59 Oral Abstract Session 08: Correlates of Protection and Exposure Tuesday, 28 October OA08.03 Systematic Analysis of HIV- Env Epitopes of Two HLA Class I Alleles Associate with Different Rates of HIV Infection in the Pumwani Sex Worker Cohort Meika EI Richmond,2, Christina A. Daniuk, Rupert E. Capina, Joshua Kimani 3, Charles Wachihi 3, Makubo Kimani 3, Thomas Bielawny, Mark G. Mendoza, T. Blake Ball,2,3, Francis A. Plummer 2,3,4, Ma Luo,2 Public Health Agency of Canada, National Lab for HIV Immunology, Winnipeg, MB, Canada, 2 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 3 University of Nairobi, Nairobi, Kenya, 4 Public Health Agency of Canada, National Microbiology Lab, Winnipeg, MB, Canada Background: HIV exposed seronegative (HESN) individuals provide a unique opportunity to study natural immunity to HIV-. Our previous studies on HESN women from the Pumwani sex worker cohort showed that A*0:0 is associated with slower seroconversion while B*07:02 is associated with rapid seroconversion. Understanding why these alleles associate with different infection rates and their epitope characteristics may provide clues for design of an effective HIV vaccine. Methods: We screened 820 peptides (9mer overlapping by 8aa) of HIV- clade A/D Env for A*0:0 and B*07:02 epitopes using itopia Epitope Discovery System. Binding eptiopes were analyzed for affinity, off-rates and validated by IFNγ ELISpot. Selected tetramers were used (A*00:VI/VK, B*0702:SL/KL) for flow cytometry analysis of CD8 T cell memory (CCR7/CD45Ra) and differentiation (CD27/CD28) phenotypes. Results: A*0:0 bound 20 epitopes, while B*0702 bound 64. No differences where seen in peptide binding affinity or off-rate between the two alleles. ELISpot responses were stronger in A*0:0+ patients (p< 0.000) and a higher proportion of A*0:0 epitopes is in the constant regions. Tetramer specific CD8s had distinct memory and differentiation profiles than non-specific CD8s (p=0.0002). Additionally, CD8 T cells specific for A*0:0 epitope YI were more likely to have an effector memory (CCR7 - CD45RA - ) phenotype than B*07:02 specific-cd8 T cells (p=0.0002). Moreover, CD8 T cells specific for both A*0:0 epitopes (YI/VK) were highly antigen experienced (CD27 - CD28 - ) compared to those specific for B*07:02 epitopes (p< 0.000). Conclusions: These data are consistent with our previous study of HIV- Gag epitopes of these alleles. A*0:0 recognizes fewer epitopes than B*07:02, suggesting narrow immune responses targeting conserved regions may be better than broad immune responses. Additionally, A*0:0 associates with higher effector memory and higher antigen experience, suggesting high cytolytic activity and potentially more effective responses. OA08.04 Comprehensive Sieve Analysis of Breakthrough HIV- Sequences in the RV44 Vaccine Efficacy Trial Paul T. Edlefsen, Morgane Rolland 2, Tomer Hertz, Sodsai Tovanabutra 2, Andrew J. Gartland, Allan C. decamp, Craig A. Magaret, Hasan Ahmed, Raphael Gottardo, Michal Juraska, Connor McCoy 3, Brendan B. Larsen 4, Eric Sanders-Buell 2, Chris Carrico 5,6, Sergey Menis 5,6, Meera Bose 2, Miguel A. Arroyo 7, Robert J. O Connell 8, Mark S. desouza 8, Sorachai Nitayaphan 7, Punnee Pitisuttithum 9, Jaranit Kaewkungwal 7, Supachai Rerks-Ngarm 0, Merlin L. Robb 2, Jason S. McLellan, Ivelin S. Georgiev, Tatsiana Kirys, Peter D. Kwong, Jonathan M. Carlson 2, Nelson L. Michael 2, William R. Schief 5,6,3, James I. Mullins 4, Jerome H. Kim 2, Peter B. Gilbert, RV44 Sequencing Team Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 US Military HIV Research Program, Silver Spring, MD, United States, 3 Fred Hutchinson Cancer Research Center, Public Health Sciences Division, Seattle, WA, United States, 4 University of Washington, Department of Microbiology, Seattle, WA, United States, 5 University of Washington, Department of Biochemistry, Seattle, WA, United States, 6 The Scripps Research Institute, IAVI Neutralizing Antibody Center and Department of Immunology and Microbial Sciences, La Jolla, CA, United States, 7 AFRIMS, Royal Thai Army Component, Bangkok, Thailand, 8 AFRIMS, US Army Component, Bangkok, Thailand, 9 Mahidol University, Faculty of Tropical Medicine, Bangkok, Thailand, 0 Thai Ministry of Public Health, Nonthaburi, Thailand, Vaccine Research Center, NIAID, NIH, Bethesda, MD, United States, 2 Microsoft Research, Redmond, WA, United States, 3 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States Background: The RV44 clinical trial showed the partial efficacy of a prime-boost pox-protein vaccine regimen with an estimated vaccine efficacy (VE) of 3% for protecting low-risk Thai volunteers against acquisition of HIV-. The impact of vaccine-induced immune responses can be investigated through sieve analysis of HIV- breakthrough infections (infected vaccine and placebo recipients). A V/V2-targeted comparison of the genomes of HIV- breakthrough viruses from vaccine and placebo recipients identified two V2 amino acid loci that differed between the vaccine and placebo groups, corroborating the finding that V2-specific antibodies in vaccine recipients were associated with a reduced risk of HIV- infection. Methods: Here we extended the V/V2 sieve analysis to the entire HIV- genome using an array of sieve analysis methods based on individual sites, k-mers and genes/proteins. Results: Across the HIV- proteome, we identified 56 amino acid sites or signatures and 9 k-mers that differed between the vaccine and placebo groups; among these, 9 signature sites and 38 k-mers were located in Envgp20, Gag, and Pro that constituted the RV44 vaccine. The nine signature sites in Env-gp20 were significantly more likely to be known antibodyassociated sites than non-signature sites (p = 0.002). In particular, one signature in V3 (37) overlapped with a hotspot of antibody recognition, and sites 369 and 424 are linked to CD4 binding site neutralization. Conclusions: Since signature sites were not preferentially restricted to the vaccine immunogens and because most of the statistical significance did not withstand multiplicity adjustment, we predict that few of the 56 genetic signatures found across the HIV- proteome are strongly linked to the RV44 vaccine-induced immune pressure. The collection of statistical methods and tools we employed constitutes an analysis platform applicable to sieve analysis of breakthrough infection genomes in general vaccine efficacy trials for diverse pathogens. ORAL ABSTRACT SESSIONS 59

60 Oral Abstract Sessions Oral Abstract Session 08: Correlates of Protection and Exposure ORAL ABSTRACT SESSIONS OA08.05 SIVsmE660 Envelope Variants from Breakthrough Infections Following DNA/ MVA Vaccination of Rhesus Macaques are Susceptible to Neutralization Stacey A. Smith, Samantha Burton, Sharmila Reddy, Katie Kilgore, Eric Hunter, Harriet Robinson 2, Rama Amara, Cynthia Derdeyn Emory University, Atlanta, GA, United States, 2 GeoVax, Inc., Atlanta, GA, United States Background: SIVmac239-based DNA prime/mva boost vaccine regimens including GM-CSF or CD40L adjuvants were tested in rhesus macaques, along with an MVA only regimen. Significant protection was observed against a low dose repeat intra-rectal SIVsmE660 challenge. We investigated whether vaccine-induced antibodies resulted in breakthrough infection by neutralization resistant variants following challenge. Methods: We identified the SIVsmE660-derived transmitted/ founder envelope (T/F Env) sequence for 4 vaccinated and 4 control monkeys using SGA. T/F Env expression vectors were used to generate pseudovirus, which was tested for neutralization sensitivity to heterologous pooled serum from chronically SIV-infected monkeys, and autologous vaccinated monkey serum collected at 3 weeks post-mva boost (3-4 weeks prior to challenge). Results: All T/F Envs were susceptible to neutralization by the chronic SIV-infected serum pools to varying degrees, but there was no significant difference between Envs from vaccinated vs. control monkeys, or between vaccine groups. In fact, the most neutralization resistant Env variants were found in two control monkeys who had no pre-challenge anti-siv immunity. All T/F Envs from vaccinated monkeys were also susceptible to neutralization by the week 3 autologous serum at IC50s of up to :3,000. Finally, a recently described signature in the Env gp20 C region, 45A/47K, did not determine resistance of the T/F Envs against heterologous or autologous neutralization. Conclusions: These results demonstrate that antibodies capable of neutralizing the autologous T/F Envs at titers exceeding those estimated to be necessary for protection against mucosal transmission were generated by these vaccination regimens. Even though the breakthrough T/F Envs were sensitive to neutralization, these animals were not protected. Thus, increasing the magnitude, breadth, and/ or local mucosal production of these antibodies may be required to enhance their protective potential. OA08.06 LB Microbicide-vaccine Combination Provides Significant Protection against Vaginal SHIV- 62P3 Challenge in Cynomolgous Monkeys Roger Le Grand, Nathalie Nathalie Bosquet, Stefania Dispinseri 2, Leslie Gosse, Delphine Des Jardins, Shen Shen 3, Georgia Tomaras 3, Nicola Hopewell 4, Susan Barnett 5, Hela Saidi, Rodolphe Thiebaut 6, Gabriella Scarlatti 2, Alethea Cope 4, Robin J. Shattock 4 Commissariat à l Energie Atomique et aux Energies Alternatives, Division of Immuno-Virology, IDMIT Center, Paris, France, 2 San Raffaele Scientific Institute, Milan, Italy, 3 Duke Human Vaccine Institute, Durham, NC, United States, 4 Imperial College, London, United Kingdom, 5 Novartis Vaccines, Cambridge, MA, United States, 6 Institut de Santé Publique, d Epidémiologie et de Développement ISPED, Bordeaux, France Background: Although a number of new biomedical prevention tools have demonstrated variable success against HIV infection in clinical trials including a partially effective microbicide (tenofovir gel) and a modestly protective vaccine (the Thai RV-44 trial), their combined introduction could provide more potent protection. This study directly explores potential positive interactions. Methods: We used a nonhuman primate model to determine whether combining a partially effective microbicide (% tenofovir gel) with an envelope based vaccine could provide enhanced efficacy against repeat intravaginal challenge with SHIV-62P3. Vaccinated animals received three nasal priming doses containing a combination of gp40 TV (Clade C) and SF62 (clade B) constructs administered with R848 (TLR7/8 agonist) followed by two intramuscular boost immunizations administered with MF59. Results: Tenofovir gel provided a 46% reduction in infection following 6 consecutive low dose intravaginal challenges (P=0.04), where tenofovir gel in vaccinated animals provided 8% reduction (P=0.02) and the vaccine alone failed to protect against SHIV-62P3 infection (P=0.85). Following 2 consecutive challenges the the combination group maintained a sustained protection of 63% (P=0.0006) while the microbicide group only provided 4% reduction in infection (P=0.02). In a second phase, protected animals were challenged a further 2 times in the absence of microbicide. Protected animals in the vaccine group initially receiving tenofovir gel when challenged in the absence of gel maintained protection (p=0.0) with a total risk reduction over 22 exposures of 38%, while animals initially receiving tenofovir gel were fully susceptible to infection. Conclusions: These important findings offer the possibility that combined implementation of new biomedical prevention strategies may provide significant reduction in HIV incidence and argues for accelerated assessment of potentially beneficial combinations through randomised controlled clinical trials. 60 HIV Research for Prevention 204 HIV R4P

61 Tuesday, 28 October Oral Abstract Session 09: Engaging, Recruiting and Retaining Trial Participants OA09.0 Why Women at High Risk for HIV- Infection Did Not Join the VOICE Study in Uganda: A Qualitative Community Study Teopista Nakyanzi, Samuel Kabwigu, Doreen Kemigisha, Sophie C. Nanziri, Patrick Ndawula, Stella Nanyonga, Juliane Etima, Flavia M. Kiweewa, Rhonda White 2, Lisa Noguchi 3, Clemensia Nakabito Makerere University Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 FHI 360, Community Program, Durham, NC, United States, 3 Microbicide Trials Network, Washington, DC, United States Background: HIV- prevalence in Uganda remains among the highest in the world, and recent surveillance data have not seen a significant drop in HIV- incidence. The VOICE study evaluated safety and effectiveness of daily use of female-controlled ARV-based oral and topical vaginal HIV prophylaxis. Despite being at risk of HIV- acquisition, and having limited prevention tools, many women declined to join the study. We describe the underlying factors that hindered at-risk women from joining VOICE. Methods: Women aged 8-45 in high-risk communities identified by the Uganda HIV/AIDS sero-behavioral Survey were sensitized about the VOICE study. Prospective participants questions and concerns were addressed, and those willing to be screened on site were systematically pre-screened using an IRB-approved checklist with multiple risk questions. Presumptively eligible women were given appointments and reminded by telephone calls. Results: From November 2009 to December 20, among the 3,27 women sensitized in the community, only 25% (n=820) were interested and pre-screened. Of those expected to turn up, only 43% (n=356) showed for appointments. Among those who did not turn up (n=464), 78% (n=362) did not know or were not sure of their partners HIV status and feared finding out their HIV status; 58% (n=269) were not living with their partner(s) and could not choose from which partner to obtain permission; and 30% (n=39) had no income and feared losing financial support from their partners. Other factors included fear of side effects, myths and misconceptions about clinical research. Others needed more time to think about participation, and the rest promised to come for screening but later changed their minds. Conclusions: Women are challenged by the fear of knowing their HIV status, exacerbated by lack of independence to make decisions about study participation. There is more need to sensitize communities about HIV prevention research, participant challenges and involving male partners in research. OA09.02 Community Engagement in a Volatile Community Post-marikana for a Phase III Microbicide Ring Trial Cheryl Emily Louw, Ntswaki Rose Masilo, Marthie de Villiers, Annalene M. Nel 2, Michelle Isaacs 2 Madibeng Centre for Research, Brits, South Africa, 2 International Partnership for Microbicides, Paarl, South Africa Background: When Madibeng Centre for Research (MCR) began its work on a Vaginal Microbicide Ring Phase III trial the surrounding community was peaceful. Wage negotiations in the surrounding mines became tense leading up to the Marikana massacre on 6 August 202, just more than 4 months into the trial. The situation in the area remained tense and also led to male partner aggression towards their female partners. Partners viewed the trial and the ring with suspicion resulting in some participants being forced to stop using the ring and to withdraw from the trial. Methods: With participant consent, MCR strategized on ways to reach out to the affected participants partners and to engage the local community: Partners were visited personally and given appropriate information on the trial Partners were invited to attend the research centre (RC) to view procedures and interview the Principal Investigator Upscaled community engagement and education occurred in the communities close to Marikana Participants were educated and equipped to deal with partners who did not wish them to use the ring, or who wanted them to withdraw against their will Results: Partners who accepted a visit from the Community Liaison Officer were in favour of the trial and their female partners participation on the trial. Some partners visited the RC to observe all procedures before accepting that their female partners could continue on the trial. Women felt more equipped to deal with partner objections to the trial. Early trial withdrawal for partner related issues decreased substantially after the intervention. Conclusions: The situation around a research centre could change dramatically overnight. Unrelated incidents could have a ripple effect on the clinical trials in the area. Partners may transfer their frustration and anger onto their female partners, affecting their compliance to the protocol. With appropriate intervention using a multi-pronged approach it is possible to turn such a situation around. ORAL ABSTRACT SESSIONS 6

62 Oral Abstract Sessions Oral Abstract Session 09: Engaging, Recruiting and Retaining Trial Participants ORAL ABSTRACT SESSIONS OA09.03 Employing a Youth-centered Approach to Investigate HIV Risk among Adolescents and Young Adults in an HIV Hyper-endemic Setting Janan Dietrich, Laura Cotton 2, Stefanie Hornschuh, Martin van der Watt, Cari L. Miller 2, Glenda Gray, Mark Brockman 2, Angela Kaida 2, on behalf of the AYAZAZI Study Team Perinatal HIV Research Unit, Soweto, South Africa, 2 Simon Fraser University, Faculty of Health Sciences, Burnaby, BC, Canada Background: Adolescents and young adults (AYA) (0-24 years) are significantly at risk of HIV acquisition across Southern Africa. The continued burden of HIV transmission among AYA reflects poor prioritization of youth-focused and engaged research yielding an inadequate understanding of intersecting factors necessary to implement accessible and effective HIV prevention programs. Use of youthcentred approaches within HIV research and programming presents a critical opportunity to engage youth, build capacity, and support youth leadership in the HIV response. Methods: AYAZAZI ( Knowing Themselves in Zulu) is a youth-centred, inter-disciplinary cohort study that aims to link socio-behavioural, structural, clinical, and biomedical data to understand HIV acquisition risk among AYA aged 6-24 years living in Soweto, South Africa. We aim to enroll a cohort of 400 AYA (HIV-negative or HIV status unknown) followed biannually for three years. Results: AYAZAZI uses a youth-centred approach to engage AYA throughout the research process. A Soweto-based Adolescent Community Advisory Board provides overall study guidance and oversight. We engaged the expertise and lived experiences of community members in the development of the survey and clinical protocol to ensure youthappropriate language and priorities. AYA research assistants from Soweto were hired and trained to support recruitment and to administer surveys. We engaged with several youth-focused community partners to conduct outreach and raise awareness of AYAZAZI. AYA research assistants will receive ongoing training to support skill development and participants will have access to regular knowledge exchange forums to support youth leadership around HIV knowledge in the community. Conclusions: Youth-centred, inter-disciplinary approaches that engage young people throughout the research process are critical to support, empower, and build local capacity required to prevent HIV infection and reverse the high HIV risk environment among youth in South Africa. OA09.04 WhatsApp!: Use of Mobile Technology in Optimizing ASPIRE Study Retention and Adherence at the Wits Reproductive Health and HIV Institute, Johannesburg Krishnaveni Reddy, Helen Rees, Thesla Palanee Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: Mobile technology is an evolving communication medium within clinical trials for participant/staff interaction and represents a personalized discrete contact platform that is available 24 hours a day. Previously, short message service (SMS) communication between staff and participants positively impacted study retention but was primarily unidirectional due to associated costs. The development of low cost instant messaging (IM) programs opens new opportunities for bidirectional communication between participants and staff. Methods: At Wits RHI, during locator information collection, most participants indicated having access to smartphones and the WhatsApp IM program. Staff then embarked on an array of strategies to utilize this communication method for retention and adherence optimization. Community health workers (CHWs) were provided with smartphones to allow continuous communication with allocated participants for visit reminders, on study related matters such as engagement activities/ events, clinic closures and the occasional check-ins. The Investigator of Record (IoR), study coordinator (SC) and clinicians also use WhatsApp to communicate with participants on clinical and study related issues discretely. Results: Participants respond to staff in real time on WhatsApp due to low costs, ease of covert use and ability to raise sensitive issues. Consequently, reporting of adverse events and adherence issues occurs and is documented and followed up as needed. In addition, participants are able to timeously communicate their challenges in attending scheduled visits so they can be re-scheduled accordingly which then impacts retention. Conclusions: The ease of use and cost efficiency of IM contributes to optimizing retention and adherence as well as adverse event reporting; factors essential in determining safety and effectiveness of the study product by allowing more frequent communication between staff and participants. 62 HIV Research for Prevention 204 HIV R4P

63 Tuesday, 28 October Oral Abstract Session 09: Engaging, Recruiting and Retaining Trial Participants OA09.05 Turning around Poor Retention: A Cape Town Experience Karen Dominguez, Katherine Gill, Leader N. Kanyiki, Ben Brown, Linda-Gail Bekker 2 Desmond Tutu HIV Foundation, Cape Town, South Africa, 2 Desmond Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town, South Africa Background: Retention in prevention trials is critical as it underpins adherence and safety monitoring. Retention requires motivation and action on behalf of participants and staff. As a site participating in a Phase III tenofovir % vaginal gel study, we experienced poor participant retention shortly after initiation in October 20. Based in a township in Cape Town, a severe increase in missed visits began when participants travelled out of the province for the December holidays. Methods: We calculated a site specific monthly retention rate using number of visits attended divided by number of visits expected. Novel ideas were urgently implemented to get participants already enrolled to return for follow-up visits. Ideas included reminder and birthday SMSes, recognition rewards for prompt and regular visits, transport for visits, as well as Saturday clinics. The enrolment process was made more stringent by instituting a pre-screen and discussion group prior to screening in an effort to ensure participant commitment. A rewards points system was designed as an incentive to come for participant visits and more points could be earned for visits on scheduled dates. The points could be accrued and redeemed for various prizes. Results: The site enrolled 53 female participants, ages Retention reached its low point with a rate of 45 %. Following and with continued use of the interventions, an average of 8% retention rate was achieved for the remainder of the year. Methods that have been more popular with the participants include the points system incentive, recognition, and vehicle transport for visits. Conclusions: Migration is a challenge for retention. Many trial sites in South Africa face this challenge. Our team has become pro-active in making retention a priority to prepare for migration and retain participants throughout the year. Through careful participant selection and use of methods to increase motivation and reduce hassle factors, retention was enhanced in this cohort. OA09.06 Reasons Boston MSM Enrolled in Placebo Controlled PrEP Trials Did Not Continue Using PrEP when it Was Available in an Open Label Study Christopher Chianese, Kenneth Mayer, Marcy Gelman, Lori Panther, Robert Grant 2,3 Fenway Health, Boston, MA, United States, 2 Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA, United States, 3 University of California, San Francisco, CA, United States Background: Boston participants who enrolled in the CDC s PrEP safety study and the iprex PrEP efficacy trial were offered open label access to tenofovir-emtricitabine (TDF/FTC) for PrEP in the iprex Open Label Extension (OLE). We identified participants who enrolled and did not take TDF/FTC at the Boston study site and evaluated study records to characterize reasons for not being administered TDF/FTC. Methods: Participants who were not dispensed TDF/FTC were identified. Visit note source documentation for each participant was analyzed to determine why participants were ineligible for drug dispensation. Reasons for drug dispensation ineligibility for each participant were then coded and sub-divided into common themes. Results: Of 79 participants who enrolled in the CDC Safety study and 87 who enrolled in iprex, 56% enrolled in iprex OLE at the Boston site between 7/7/20 and 3/29/202. Median age was 44; 65% white, 23% Black, 2% other. Of 93 participants enrolled, 37% were not dispensed TDF/FTC. Median age of participants not dispensed TDF/FTC was 47; 59% white, 35% Black, 6% other. Of 34 participants, 38% did not perceive they had high enough risk for HIV to justify PrEP; 8% were concerned about side effects; 5% had active clinically significant medical problems (including cardiac disease, pulmonary disease, DM requiring hyperglycemic medication); % were taking contraindicated medications; 6% had seroconverted after completing the CDC Safety study, 6% felt pill-taking would be too burdensome; 6% indicated it was because of personal choice but did not provide reasons. Conclusions: Because PrEP study participants changed clinically and behaviorally over time, these findings demonstrate that ongoing clinical examination and behavioral assessment of PrEP users is important, and PrEP delivery should not be automatic. Additional exploration into contextual factors that influence an individual s decision to take PrEP is needed as PrEP becomes one of several HIV prevention modalities. ORAL ABSTRACT SESSIONS 63

64 Oral Abstract Sessions Oral Abstract Session 0: Bacterial Vaginosis and HSV-2: Impact on Genital Immunity ORAL ABSTRACT SESSIONS OA0.0 The Effects of Hormones and Vaginal Microflora on the Content of MUC, MUC4, MUC5AC and MUC7 in the Cervicovaginal Fluid (CVF) Bernard J. Moncla,2, Catherine Chappell, Brian M. Debo 2, Ingrid S. Macio 3, Katherine E. Bunge, Sharon L. Hillier,2 University of Pittsburgh, Ob/Gyn and Reproductive Sciences, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Microbiology, Pittsburgh, PA, United States, 3 Magee-Womens Research Institute, Clinical Research, Pittsburgh, PA, United States Background: Mucins are glycoproteins that protect and lubricate epithelial surfaces. In humans, they can be secreted (MUC5AC, MUC7) or membrane bound (MUC, MUC4, MUC6). Mucins can trap HIV but little is known about the impact of the vaginal microbiota and hormonal status on the mucin content in vaginal and cervical fluid. The objective of this study was to measure the quantity of 5 MUCs in the CVF of women under different hormonal conditions, stratifying for vaginal microflora. Methods: CVF was collected via catamenial cup from 65 healthy asymptomatic reproductive aged women in the follicular (n=27) or proliferative (n=26) phase, women using levonogestrol (LNG) IUDs (n=28), DMPA (n=29) or combined oral contraceptives (n=27) and 29 post-menopausal women. Vaginal smears were evaluated using the Nugent criteria for bacterial vaginosis (BV) among reproductive age women. The MUC content of the samples was evaluated using ELISA. Student s t-test and one-way analysis of variance with post-hoc comparisons made using Bonferroni s multiple comparisons procedure were used to assess statistical significance. Results: MUCs, 4, 5AC and 7 were detected at significantly higher concentrations in CVF with increasing Nugent score (P< 0.05), while MUC6 was not linked to BV status. By contrast, MUC content was minimally impacted by hormonal status. There was no statistically significant difference in MUC and MUC5AC among women in the different hormonal groups. MUC4 was significantly increased among post-menopausal women (P=0.005, comparing post-menopausal women to all others), and women using a LNG IUD had lower levels of MUC7 in the CVFs compared to other reproductive age women (P=0.006). Conclusions: In this study, vaginal microflora had a greater impact on MUC content of CVF than hormonal status. Women with BV have increased levels of secreted and membrane bound mucins, indicating that microbiota induce changes in the expression of cervical mucins which could contribute to an increased risk of HIV. OA0.02 HSV-2-driven Changes in α 4 β 7 Expression Correlate with Increased Susceptibility to SHIV ex vivo and in vivo Diana Goode, Rosaline Truong, Meropi Aravantinou, James Blanchard 2, Agegnehu Gettie 3, Melissa Robbiani, Elena Martinelli Population Council, New York, NY, United States, 2 Tulane National Primate Research Center, Tulane, LA, United States, 3 Aaron Diamond AIDS Research Center, New York, NY, United States Background: The availability of highly susceptible HIV target cells that can rapidly reach the mucosal lymphoid tissues may increase the chances of an otherwise rare transmission event to occur. α 4 β 7 high CD4 + T cells are highly susceptible to HIV infection, they home specifically to the GALT and HIV-gp20 interacts with α 4 β 7. We hypothesized that HSV- 2, which augments the risk of HIV acquisition, modulates the expression of α 4 β 7 and other homing molecules in the vaginal tissue increasing its susceptibility to HIV and SIV. Methods: To test this hypothesis we used an in vivo rhesus macaque (RM) model of HSV-2 vaginal infection and we developed a new ex vivo model of macaque vaginal explants. Results: In vivo, we found that HSV-2 chronically infected RMs were more susceptible to vaginal SHIV SF62P3 infection. Ex vivo HSV-2 infection increased the susceptibility of vaginal tissue to SHIV. Notably, in vivo HSV-2 infection decreased expression of α 4 β 7 on blood α 4 β 7 high CD4 + T cells. In uninfected RMs the frequency of these cells inversely correlated in blood and vaginal tissue. Ex vivo HSV-2 infection of vaginal explants increased the frequency of α 4 β 7 high CD4 + T cells and α 4 β 7 high CD80 + DCs in the mucosa and this directly correlated with HSV-2 replication. HSV- 2 also increased the frequency of CD03 + CD4 + T cells, however, this negatively correlated with HSV-2 replication. The expression of α 4 β 7 on migratory CD4 + T cells and the frequency of CD62L + CD4 + T cells also correlated with HSV-2 replication. Importantly, the HSV-2-driven increase in the frequency of α 4 β 7 high CD4 T directly correlated with SHIV replication in the HSV-2 infected tissues. Conclusions: Our results suggest that HSV-2 infection modulates the expression of homing molecules. In particular the HSV-2-driven increase in the availability of α 4 β 7 high CD4 + T cells and DCs correlates with increased susceptibility to SHIV infection. Thus, blocking a 4 b 7 may decrease the risk of HIV vaginal acquisition in HSV-2 infected and potentially HSV-2 uninfected women. 64 HIV Research for Prevention 204 HIV R4P

65 Tuesday, 28 October Oral Abstract Session 0: Bacterial Vaginosis and HSV-2: Impact on Genital Immunity OA0.03 Incident Herpes Simplex Virus Type 2 Associated with HIV Infection in Phambili James G. Kublin, Zoe Moodie 2, Barbara Metch 2, Linda-Gail Bekker 3, Gavin Churchyard 4, Maphoshane Nchabeleng 5, Koleka Mlisana 6, Mary Allen 7, Larry Corey 8, Glenda Gray 9, on behalf of the HVTN 503 Study Team HIV Vaccine Trials Network, Seattle, WA, United States, 2 FHCRC, SCHARP, Seattle, WA, United States, 3 University of Cape Town, The Desmond Tutu HIV Centre, Cape Town, South Africa, 4 Aurum Institute for Health Research, Klerksdorp, South Africa, 5 Medunsa HIV Research Unit, Medunsa, South Africa, 6 University of KwaZulu Natal, Durban, South Africa, 7 DAIDS, NIAID, NIH, Vaccine Clinical Research Branch, Vaccine Research Program, Rockville, MD, United States, 8 FHCRC, HVTN, Seattle, WA, United States, 9 Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital, Soweto, South Africa Background: In the Phambili preventative HIV- vaccine efficacy trial conducted in South Africa, herpes simplex virus type 2 (HSV2) infection at enrollment increased the risk of HIV- acquisition in men but not in women. We monitored for incident HSV2 and potential associations with HIV infection. Methods: HSV2 prevalence at enrollment and incidence at 24 months on study were determined by validated western blot assay. HIV- testing occurred at specified intervals and RNA PCR was used to confirm HIV- infection. Multivariate logistic regression modeling was used to investigate associations. Results: HSV2 prevalence at enrollment was 3% (248/798; 95% CI 28-34%); prevalence of HSV2 was significantly higher in women (49%) than in men (6%); p< Prevalence increased with age for both genders, with 45% of men and 75% of women 27 years or older infected. Prevalence varied by site, but not by circumcision status after adjustment for age. HSV2 incidence at 24 months was % (43/384; 95% CI 8-4%); acquisition of HSV2 was significantly higher in women (20%) than in men (7%); p< Incident HSV2 infection among men was positively associated with age (OR=.2 per year increase) and negatively associated with smoking dagga (OR=0.2) and being apart regularly from a main partner (OR=0.2). Among women, having a casual/anonymous partner within 6 months prior to screening was the only significant factor associated with incident HSV2 infection (OR=3.6). Sixty HIV- infections occurred within participants first 24 months in study, of whom 30 were HSV2 positive at enrollment. Among the other 30, were diagnosed as HSV2 positive prior to or concurrent with HIV infection diagnosis; 9 were HSV2 negative at the time of HIV- diagnosis. By 24 months, HIV infection was associated with HSV2 infection, adjusted for age, both for men (OR=5.6) and for women (OR=3.7). Conclusions: HSV2 acquisition is markedly associated with HIV- infection in both heterosexual men and women in South Africa. OA0.04 Mucosal Integrity Factors Are Perturbed during Bacterial Vaginosis: A Proteomic Analysis Kelly Arnold, Kenzie Birse 2, Lyle Mckinnon 3, Jiriam Lingappa 4, Rick Novak 5, Garrett Westmacott 6, T. Blake Ball 6, Doug Lauffenburger, Adam Burgener 6 MIT, Boston, MA, United States, 2 University of Manitoba, Winnipeg, MB, Canada, 3 CAPRISA/University of KwaZulu Natal, Durban, South Africa, 4 University of Washington, Seattle, WA, United States, 5 University of Illinois, Chicago, IL, United States, 6 Public Health Agency of Canada, Winnipeg, MB, Canada Background: Mucosal inflammation, such as is found during bacterial vaginosis (BV), has been associated with increased risk of HIV infection in women. However, the mechanisms and/or drivers of HIV-susceptibility are not well understood. Here we utilized a global proteomics approach, coupled to multivariate computational modeling, to examine impact of BV on the female genital tract mucosa in two distinct populations. Methods: Cervicovaginal lavage samples collected from North American individuals (Group A: BV+, n=8; BV-, n=26) and Kenya (Group B: BV+, n=0; BV-, n=0) were analyzed by tandem mass spectrometry. Data was analyzed using a combination of hierarchical clustering, pathway analysis, and multivariate (Lasso) modeling. Results: Approximately 650 host (human) and 00 bacterial protein factors were identified in mucosal samples. BV+ individuals showed significantly affected host protein expression including 52 in group A and 92 in group B (p< 0.05). Microbiome proteins were also affected, including significant reduction of Lactobacillus sp. factors (8, p< 0.05) and elevation of numerous (>20 species) anaerobic bacterial proteins (50) in BV (p< 0.05). Hierarchical clustering revealed two distinct branches of upregulated/downregulated protein groups. Functional enrichment analysis of downregulated clusters revealed that cornified envelope (Grp A:.2E-3; B: 6.6E-6) and keratinization (A:.8 E-4; B:.E-4) tissue integrity factors were common to both datasets. Lasso multivariate modeling further identified 4 specific biomarkers (distinguishing BV+ individuals with 90% accuracy) that associated with increased host metabolism, membrane integrity, and toll-like receptor signaling. Conclusions: These data suggest that BV-associated inflammation affects tissue integrity factors in the vaginal compartment, and may help to explain BV-related HIV infection. ORAL ABSTRACT SESSIONS 65

66 Oral Abstract Sessions Oral Abstract Session 0: Bacterial Vaginosis and HSV-2: Impact on Genital Immunity ORAL ABSTRACT SESSIONS OA0.05 Effect of Bacterial Vaginosis on Markers of Genital Tract Inflammation and Mucosal Immunity: Mechanisms for Susceptibility to HIV Infection Andrea Ries Thurman, Thomas D. Kimble, Tina D. Cunningham 2, Betsy C. Herold 3, Pedro Mesquita 3, Ashley Huber 3, Raina N. Fichorova 4, Hassan Y. Dawood 4, Titilayo Fashemi 4, Neelima Chandra, Jill L. Schwartz, Gustavo F. Doncel CONRAD, Eastern Virginia Medical School, Norfolk, VA, United States, 2 Eastern Virginia Medical School, School of Public Health, Norfolk, VA, United States, 3 Albert Einstein College of Medicine, Infectious Diseases and Pediatrics, Bronx, NY, United States, 4 Brigham and Women s Hospital, Harvard Medical School, Laboratory of Genital Tract Biology, Boston, MA, United States OA0.06 LB Association of Tenofovir (TFV) Detection with Reduced Risk of Herpes Simplex virus Type-2 (HSV-2) Acquisition in the VOICE (MTN 003) Study Jeanne Marrazzo, Lorna Rabe 2, Cliff Kelly 3, Edward Livant 2, Z. Mike Chirenje 4, Barbra Richardson 3, James Dai 3, Jeanna Piper 5, Sharon Hillier 2, the VOICE Study Team University of Washington, Seattle, WA, United States, 2 Magee Women Research Institute, Pittsburgh, PA, United States, 3 SCHARP - FHCRC, Seattle, WA, United States, 4 University of California San Francisco Research Programme, Harare, Zimbabwe, 5 DAIDS, NIAID, NIH, Bethesda, MD, United States Background: Bacterial vaginosis (BV), a highly prevalent alteration of vaginal flora, has been linked to an increased risk of HIV acquisition and transmission in observational studies, but the underlying biologic mechanisms are not known. We measured markers of subclinical vaginal inflammation, endogenous antimicrobial activity, and vaginal flora in women with BV and repeated sampling week and month after completion of metronidazole therapy. Methods: Longitudinal, open label study (ClinicalTrials.gov #NCT ) of 33 women with a Nugent score of 4 or higher. All women had genital swabs, cervicovaginal (CV) fluid lavage (CVL) and vaginal biopsies obtained at enrollment (active BV) and received 7 days of metronidazole treatment. Repeat sampling was performed approximately week and month after completion of therapy. Results: The CVL from women with resolved BV (Nugent 0-3) had significantly higher anti-hiv activity compared to women with abnormal microbiota (Nugent 4-0). The mean anti-hiv activity of women with BV was < 0, indicating that their CVL enhanced HIV infection of TZMbl cells in vitro. Genital tract interleukins IL-β and ICAM- levels were significantly higher (p < 0.05) and SLPI levels were lower (p = 0.02) in the presence of BV compared to normal microbiota. Anti- HIV activity of CVL was significantly associated with the Nugent score and genital IL-β and IL-8 levels in a multivariate model controlling for participant race and visit. Although women with BV had significantly lower numbers of CD45+, CD3+ and CD8+ vaginal immune cells compared to racially matched women without BV, they had significantly greater numbers of CCR5+ cells and increased numbers CD4+ cells and CD4/CD8 ratios. Conclusions: These data support that BV is associated with changes in select soluble immune mediators, an increase in mucosal HIV target cells and a reduction in endogenous CV antiviral activity, which may contribute to the increased risk of HIV acquisition. Background: In sub-saharan Africa, HSV-2 infection is common, and increases risk of HIV transmission and acquisition. TFV gel applied before and after vaginal intercourse provided partial protection from HSV-2 acquisition in CAPRISA 004. We assessed efficacy of % vaginal TFV gel in preventing HSV-2 acquisition in women in the VOICE Study, a 5-arm, randomized, double-blind, placebo-controlled trial that studied daily use of oral and vaginal tenofovir for HIV- pre-exposure prophylaxis. Methods: From September 2009-June 20, 5,029 women were enrolled in VOICE in South Africa, Uganda, and Zimbabwe.,004 women were randomized to receive TFV % gel. Testing for HSV-2 typespecific antibody (Focus HerpeSelect EIA) was performed on plasma from enrollment and study exit (positive result index value >3.5). TFV measured in plasma (first quarterly visit) or vaginal swab (6 month visit) was used as a measure of gel use. Using Poisson regression, we analyzed the association between TFV detection in these samples and HSV-2 acquisition. Results: Of women in the TFV gel arm, 566 were HSV-2 uninfected at enrollment (56%); of these, 53 (94%) had first quarter plasma TFV and end-of-study HSV-2 serology available. Over follow-up of 5 PY, 92 incident HSV-2 cases occurred: 77 in women with no plasma TFV detected and 5 in those with plasma TFV (incidence 20.2 (95% CI ) vs.6 (6.5-9.), respectively). Plasma TFV detection was associated with reduced risk of HSV-2 seroconversion (unadjusted incidence rate ratio (IRR) 0.57 ( ; P=0.049); IRR adjusted for country, age, marital status, 2 sex partners, hormonal contraception type, anal sex, and HIV status 0.54 ( ; P=0.039)). With TFV detection in vaginal swabs, adjusted IRR was 0.7 (0.45-.; P=0.22). Conclusions: Detection of plasma TFV among TFV gel users was associated with reduced risk of HSV-2 acquisition after controlling for potential risk modifiers, including sexual behavior and HIV- acquisition. 66 HIV Research for Prevention 204 HIV R4P

67 Oral Abstract Session : Vaccine Development: Emerging Insights Wednesday, 29 October OA.0 OA.02 African Early Infection Cohort as a Platform for Vaccine Discovery: The IAVI Protocol C Experience Jill Gilmour, Anatoli Kamali 2, Etienne Karita 3, William Kilembe 4, Eduard J. Sanders 5, Omu Anzala 6, Susan Allen 7, Vinodh Edward 8, Fran Priddy 9, Matt A. Price 9, Gladys Macharia 5, Joshua Baalwa 0, Shane Crotty, Thomas Denny 2, Elise Landais, Persephone Borrow 3, Jianming Tang 0, Michael Busch 4, Jessica Prince 7, Dan Claiborne 7, Pascal Poignard, Pat Fast 9, Eric Hunter 7 IAVI Human Immunology Lab, London, United Kingdom, 2 MRC/ UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 3 Project San Francisco, Kigali, Rwanda, 4 ZEHRP, Lusaka, Zambia, 5 CGMRC-KEMRI, Kilifi, Kenya, 6 KAVI-Institute of Clinical Research, University of Nairobi, Nairobi, Kenya, 7 Emory University, Atlanta, GA, United States, 8 Aurum Institute, Rustenburg, South Africa, 9 IAVI, New York, NY, United States, 0 UAB, Birmingham, AL, United States, La Jolla Institute for Allergy and Immunology, La Jolla, CA, United States, 2 Duke Human Vaccine Institute, Durham, NC, United States, 3 University of Oxford, Oxford, United Kingdom, 4 Blood Systems Research Institute, San Francisco, CA, United States Background: Early HIV host-virus interactions provide important clues to vaccine discovery and conducting acute infection studies in Africa with multiple clades is vital to development of a global vaccine. Methods: We followed cohorts at high risk of HIV acquisition at 9 research centers in East and Southern Africa to identify volunteers with incident HIV infection. Infection date was estimated from serial antibody, p24 and/or PCR results. Volunteers were invited to bring a sexual partner for visit. Follow up was monthly for the first 3 months post-infection, quarterly through 2 years, and every 6 months thereafter. At every visit, volunteers had a symptom-directed exam including medications, and blood for viral load, CD4+ T cell counts and PBMCs. Results: From , 64 seroconverters and 406 sexual partners were enrolled. Follow up is ongoing; to date over 2,500 person years of study time has provided a valuable platform for characterizing the HIV genetic bottleneck at transmission and founder virus characteristics. T cell responses in the acute phase have been shown to be associated with virus control and viral replicative capacity with both set point viral load and CD4+ T cell decline. Disease progression varied by sub-type, with subtype-c and D-infected volunteers progressing faster than A, and HLA alleles associated with outcomes have been characterized, such as B*44, B*45, B*57 and B*8. The development and isolation of neutralizing antibodies 2-3 years post infection is being described and the level of circulating memory T follicular helper cells has been associated with the development of such antibodies. Sample panels have provided valuable to the development of pseudovirions and validating assays to detect recent infection. Conclusions: Prospective data and samples from early infection cohorts are increasingly valuable for vaccine discovery work. Identifying host and viral factors associated with acquisition and early viral control across clades can inform the design of a global HIV vaccine. Intradermal HIV-DNA Given with or without Intradermal Electroporation Is Safe and Highly Immunogenic in Healthy Swedish HIV- DNA/MVA Vaccinees Charlotta Nilsson,2,3, Bo Hejdeman 4, Karina Godoy-Ramirez, Teghesti Tecleab, Gabriella Scarlatti 5, Andreas Bråve,2, Patricia L. Earl 6, Richard R. Stout 7, Merlin L. Robb 8, Robin Shattock 9, Gunnel Biberfeld,2, Eric Sandström 4, Britta Wahren 2 The Public Health Agency of Sweden, Solna, Sweden, 2 Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 3 Karolinska Institutet, Department of Laboratory Medicine, Huddinge, Sweden, 4 Södersjukhuset and Karolinska Institutet, Venhälsan and Department of Education and Clinical Research, Stockholm, Sweden, 5 IRCCS San Raffaele Scientific Institute, Milan, Italy, 6 National Institutes of Health, National Institute of Allergy and Infectious Diseases, Bethesda, MD, United States, 7 Bioject Inc., Tualatin, OR, United States, 8 Walter Reed Army Institute of Research, Department of Retrovirology, Rockville, MD, United States, 9 Imperial College, Department of Infectious Diseases, Division of Medicine, London, United Kingdom Background: DNA-based vaccines have been shown to be safe but weakly immunogenic in humans. We compared safety and immunogenicity of intradermal (id) vaccination with and without electroporation (EP) in a phase I randomized placebo-controlled trial of a HIV-DNA prime HIV-MVA boost vaccine in healthy Swedish volunteers Methods: Twenty-five volunteers were randomized to receive HIV-DNA plasmid vaccine 0.6 mg id using a needle-free device (Zetajet) with (n=6) or without id EP (Dermavax) (n=9). An additional five volunteers were placebo recipients. HIV-DNA plasmids encoding Env gp60 subtypes A, B, and C; rev B; Gag A and B and RTmut B were given at weeks 0, 6 and 2. Boosting with recombinant MVA-CMDR expressing HIV- Env, Gag and Pol of CRF0_AE (HIV-MVA) or placebo was performed at weeks 24 and 40. Nine of the vaccinees received a subtype C CN54 gp40 protein boost in combination with HIV-MVA. Results: The intradermal EP delivery was very well tolerated. After three HIV-DNA immunizations, the IFN-γ ELISpot response rate to Gag was slightly higher in HIV-DNA EP recipients (5/5, 33%) than in HIV-DNA id recipients (/7, 4%, p=0.658). The first HIV-MVA or HIV-MVA+gp40 vaccination increased the IFN-γ ELISpot response rate to an overall 8/9 (95%). CD4 + and/or CD8 + T cell responses to Gag or Env were demonstrable in 94% of vaccinees. A balanced CD4 + and CD8 + T cell response was noted, with 78% and 7% responders, respectively. IFN-γ and IL-2 production dominated the CD4 + T cell response to Gag and Env. The CD8 + response to Gag was broader with expression of IFN-γ, IL-2, MIPβ and CD07. No differences were seen between groups. Binding antibodies were induced after the second HIV-MVA+/-gp40 in 93% of vaccinees to subtype C Env. The highest titers were seen among EP/ gp40 recipients. Conclusions: Intradermal electroporation of HIV-DNA was well tolerated. Strong cell-mediated and antibody-mediated immune responses were elicited by the HIV-DNA priming and HIV-MVA boosting regimen, irrespective of id EP use. ORAL ABSTRACT SESSIONS 67

68 Oral Abstract Sessions Oral Abstract Session : Vaccine Development: Emerging Insights OA.03 VLP-expressing DNA/MVA Vaccines: The Effect of Schedule and Regimen on Antibody Magnitude and Avidity OA.04 Vaccine Enhancement Confirmed among Men in HVTN 503-S, Final Results from a Recall Study of Phambili Participants ORAL ABSTRACT SESSIONS Susan P. Buchbinder,2, Christine Hay 3, Nicole Grunenberg 4, Paul Goepfert 5, Tomaras Georgia 6, Kelly Seaton 6, Alicia Sato 7, Marnie Elizaga 4, Xuesong Yu 7, Peter Gilbert 7, Bernard Moss 8, Harriet Robinson 9 San Francisco Department of Public Health, Bridge HIV, San Francisco, CA, United States, 2 University of California at San Francisco, Medicine, Epidemiology and Biostatistics, San Francisco, CA, United States, 3 University of Rochester Medical Center, Rochester, NY, United States, 4 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 5 University of Alabama-Birmingham, Birmingham, AL, United States, 6 Duke Human Vaccine Institute, Durham, NC, United States, 7 SCHARP- FHCRC, Seattle, WA, United States, 8 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 9 GeoVax, Inc., Smyrna, GA, United States Background: HIV- vaccines ideally induce high magnitude and avidity antibody (Ab) responses to the envelope glycoprotein (Env). In particular, the highly conserved immunodominant region (IDR) of gp4 is an important target for virion capture and ADCC. We compared Ab responses between different regimens and schedules in HIV negative healthy participants (pts) to evaluate the addition of a 3rd MVA dose, spacing between MVA doses, and the inclusion of co-expressed GM-CSF on Ab magnitude and avidity. Methods: The JS7 (D) and GEO-D03 DNA (Dg) and HIV62 MVA (M) vaccines produce virus-like particles displaying trimeric, membranebound Env; the GEO-D03 DNA co-expresses GM-CSF. HVTN 205 enrolled 50 pts in DDMM (0,2,4,6 mo.) and 75 pts in an MM_M (0,2,6 mo.) regimens ; HVTN 094 enrolled 5 pt each in DgDgMM_M (0,2,4,6,0 mo) and DgDgM_M (0,2,4,8 mo.) regimens. Results: All regimens were safe and well tolerated. Two MVA doses generated IgG to IDR gp4 in 73%-00% of participants. The 3 rd MVA dose significantly increased the magnitude (HVTN 205) and avidity (HVTN 205 and 094) of this IgG. This response was durable, with 58% of pts still positive 6 months after the 3 rd MVA dose in HVTN 205, and the median magnitude among persistent responders declining only ½ log. Increased spacing between the st and 2 nd MVA inoculations showed significantly enhanced avidity but no difference in magnitude or response rates. Including co-expressed GM-CSF in the DNA prime did not enhance the magnitude or avidity of the Env-specific Ab. At the end of the immunization regimen in HVTN 094, median avidity indices were 34 and 23 for the 3- and 2-dose MVA arms. Conclusions: The addition of a spaced 3 rd MVA boost to a DDMM regimen enhances the magnitude and avidity of the Env-specific Ab response, and appears to be durable 6 months after immunization without additional boosting. ADCC and virion capture assays are being run to evaluate the unique immune profile and promise of this regimen for protection against clade B infection. Zoe Moodie, Barbara Metch, Mary Allen 2, Linda-Gail Bekker 3, Gavin Churchyard 4, Maphoshane Nchabeleng 5, Koleka Mlisana 6, James Kublin, Glenda E. Gray 7,8 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Vaccine Research Program, Division of AIDS, Bethesda, MD, United States, 3 University of Cape Town, Desmond Tutu HIV Foundation, Cape Town, South Africa, 4 Aurum Institute for Health Research, Johannesburg, South Africa, 5 University of Limpopo, Medunsa, MEDUNSA HIV Clinical Research Unit, Mankweng-E, South Africa, 6 University of KwaZulu-Natal, Centre for AIDS Programme for Research in South Africa, Durban, South Africa, 7 University of the Witwatersrand, Perinatal HIV Research Unit, Johannesburg, South Africa, 8 South African Medical Research Council, Cape Town, South Africa Background: Final analysis of the Phambili study, assessing the MRK Ad5 gag/pol/nef subtype B preventive HIV- vaccine in South Africa, showed a higher rate of HIV- infection in the vaccine group compared to placebo after 42 months of follow-up, with a statistically significant difference among men, irrespective of circumcision or Ad5 serostatus, but not amongst women. As most follow-up occurred after participant unblinding, results may have been influenced by ascertainment bias related to dropout. Consequently, HVTN 503-S was initiated in June 203 to recall all uninfected participants for additional HIV testing. Methods: Phambili participants who were HIV uninfected at last Phambili contact were contacted for a follow-up visit which included HIV testing, risk prevention counseling, behavioral risk assessment and, for men, circumcision assessment. Cox proportional hazards models were used to estimate the vaccine:placebo hazard ratio (HR) from time of enrollment in Phambili. Results: Overall, 464 (67%) of the 695 uninfected Phambili participants were enrolled in 503-S, with no differences in contact disposition between treatment or gender groups. Sites were unable to contact 95 (4%). Among the 87 participants who dropped out of Phambili, 37% enrolled. The median time from last Phambili HIV test to 503-S testing was 33 months (range 27-77). For women, 8 new infections were diagnosed among vaccinees (n=99) and 4 among placebos (n=06). Among men, new infections were diagnosed in the vaccine group (n=30) and 3 among placebos (n=29). The Phambili HR was 2.46 (95% CI ) among men, adjusted for baseline HSV-2; with the additional 503-S follow-up, it was 2.75 (.49, 5.06). new infections were diagnosed among Phambili dropouts. Conclusions: Sites successfully located and scheduled participants despite more than two years since participants last Phambili visit. HIV testing results supported the conclusion of vaccine enhancement of HIV- acquisition among men. 68 HIV Research for Prevention 204 HIV R4P

69 Oral Abstract Session : Vaccine Development: Emerging Insights Wednesday, 29 October OA.05 OA.06 LB HIV-specific Antibody in Rectal Secretions Following Late Boosts in RV44 Participants (RV305) Siriwat Akapirat, Chitraporn Karnasuta, Sirinan Madnote, Hathairat Savadsuk, Jiraporn Puangkaew, Surawach Rittiroongrad, Sandhya Vasan, Viseth Ngauy, Merlin L. Robb 2,3, Jean-Louis Excler 2,3, Punnee Pitisutthithum 4, Supachai Rerks- Ngarm 5, Nelson L. Michael 3, Mark S. de Souza 6,7, Jerome H. Kim 3, Robert J. O Connell, Nicos Karasavvas, on behalf of the RV305 Study Group Armed Forces Research Institute of Medical Sciences, Department of Retrovirology, Bangkok, Thailand, 2 Henry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 3 Walter Reed Army Institute of Research, U.S. Military HIV Research Program, Silver Spring, MD, United States, 4 Mahidol University, Faculty of Tropical Medicine, Bangkok, Thailand, 5 Ministry of Public Health, Department of Disease Control, Nonthaburi, Thailand, 6 Cooper Human Systems, Nashua, NH, United States, 7 National Institute of Infectious Diseases, AIDS Research Center, Tokyo, Japan Background: Rectal mucosa is the primary site of HIV acquisition during anal intercourse. Anti-HIV envelope antibodies were identified as a correlate of risk in the RV44 efficacy trial but the presence of vaccine-induced antibodies in rectal secretions has not been previously assessed. Methods: HIV-specific antibodies in rectal secretions were measured. HIV uninfected RV44 male vaccinees (n=26) were randomized to receive late boosts (RV305) consisting of 2 injections of ALVAC-HIV and/or AIDSVAX B/E or placebo at 0 and 6 months. Rectal secretions collected with Merocelâ sponges. Total and HIV envelope gp20 A244gD specific IgG and IgA antibodies were measured by ELISA at study entry and 2-week post first and second vaccinations. Results: Total IgG and IgA were detected in 97% ( µg/ml) and 86% of samples ( µg/ml), respectively, throughout the three times points. Anti-A244gD IgG responses were undetectable in ALVAC- HIV/AIDSVAX B/E and AIDSVAX B/E groups, but one sample in ALVAC- HIV group had a low level of anti-a244gd IgG at baseline. Two weeks post first injection, anti-a244gd IgG was present in 70% and 57% ( and µg/ml) of ALVAC-HIV/AIDSVAX B/E and AIDSVAX B/E groups, respectively. Post second injection, response rates decreased to 33% in the ALVAC-HIV/AIDSVAX B/E group ( µg/ml) and to 29% in the AIDSVAX B/E group ( µg/ml). No responses were detected in the ALVAC-HIV group. IgA antibodies to A244gD were not detected in any of the tested samples. Conclusions: IgG antibodies to A244gD were detectable in rectal secretions in the majority of RV44 vaccine recipients who received late boosts with ALVAC-HIV/AIDSVAX B/E or AIDSVAX B/E implying a possible mechanism for vaccine protection following anal HIV exposure. IgA responses to HIV gp20 were absent from all vaccination groups despite total IgA being higher than total IgG. Merocelâ sponges are effective in collecting rectal secretions for antibody measurement. HVTN 097: Evaluation of the RV44 Vaccine Regimen in HIV Uninfected South African Adults Glenda E. Gray,2, Erica Andersen-Nissen 3, Nicole Grunenberg 4, Ying Huang 5, Surita Roux 6, Fatima Laher 7, Craig Innes 8, Niya Gu 9, Carlos DiazGranados 9, Sanjay Phogat 9, Carter Lee 0, Edith Swann, Jerome Kim 2, Robert O Connell 2, Nelson Michael 2, Britta Flach 3, Steve DeRosa 3, Nicole Frahm 3, Lynn Morris 4, David Montefiori 5, Peter Gilbert 5, Georgia Tomaras 5, Julie McElrath 3,3, Lawrence Corey 4, HVTN 097 South African Medical Research Council, Cape Town, South Africa, 2 Perinatal and HIV Research Unit, Soweto, South Africa, 3 Cape Town HVTN Immunology Laboratory/Hutchinson Centre Research Institute, Cape Town, South Africa, 4 Fred Hutchinson Cancer Research Center, HVTN, Seattle, WA, United States, 5 Fred Hutchinson Cancer Research Center, SCHARP, Seattle, WA, United States, 6 Desmond Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town, South Africa, 7 Perinatal HIV Research Unit,Faculty of Health Sciences, University of the Witwatersrand, Soweto, South Africa, 8 Aurum Institute for Health Research, Klerksdorp HVTN CRS, Klerksdorp, South Africa, 9 Sanofi Pasteur, Swiftwater, PA, United States, 0 Global Solutions for Infectious Diseases, South San Francisco, CA, United States, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 2 U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD, United States, 3 Fred Hutchinson Cancer Research Center, HVTN Laboratory Program, Seattle, WA, United States, 4 National Institute for Communicable Diseases of the NHLS, Johannesburg, South Africa, 5 Duke University Human Vaccine Institute and the Center for HIV/AIDS Vaccine Immunology, Duke University School of Medicine, Durham, NC, United States Background: Following the RV44 trial demonstration of 3% vaccine efficacy in preventing HIV infection in Thailand, HVTN 097 was designed to evaluate the same regimen in South Africans to ascertain whether their immune response profiles were similar to Thais. This study was conducted in preparation for evaluating a similar clade C HIV vaccine regimen in South Africa. Our study was critical as previous studies have demonstrated that age, gender and BMI impact vaccineinduced immune responses. Methods: ALVAC-HIV (vcp52) expressing HIV- Env (clade E, gp20 from strain 92TH023 and clade B TM gp4 from strain HIV Lai, and Gag and protease (clade B) was administered at baseline then, 3 and 6 months later with AIDSVAX B/E, bivalent HIV gp20 subtypes B (MN) and E (A244) adsorbed to alum at the last 2 vaccinations. Immune responses were measured 2 weeks after the last immunization. Intracellular Cytokine Staining identified response rates and frequencies of HIV-specific T cells expressing IFN-γ and/or IL-2. Binding antibodies to HIV- gp20 and VV2, IgG subclass, and antibody functions (ADCC, avidity index, nab, and virion capture) are ongoing. Immune responses were stratified by age, gender and BMI. Results: In 68 participants, overall peak response rates of Env-specific CD4+ T cells expressing IFN-γ and/or IL-2 was 70.6% (95% CI 58.9%, 80.%), similar to or greater than responses in RV44. Although not significant, participants < 25 years old had higher response rates observed than those 26 (76.0% vs. 55.6%, p=0.08). Response rates in females (75%; 95%CI 56.6%, 87.3%) were similar to males (67.5%; 95%CI 52.0%, 79.9%). Response rates stratified by BMI categories of < 25, 25-30, >30 were 63.6%, 82.4% and 85.7%, respectively (p< 0. for BMI 25). Conclusions: Response rates and magnitudes of Env-specific CD4+ T cells in South Africans induced by the same vaccine regimen used in RV44 were at least comparable to or better than those induced in RV44. Age/gender or BMI did not affect CD4+ T-cell response rates. ORAL ABSTRACT SESSIONS 69

70 Oral Abstract Sessions Oral Abstract Session 2: Towards Broadly Neutralizing Antibody Induction OA2.0 Maturation of Broadly Neutralizing VV2- directed Antibodies in the Context of Autologous Viral Escape OA2.02 Development of a V/V2-targeting Quaternary-specific Broadly Neutralizing Lineage ORAL ABSTRACT SESSIONS Jinal N. Bhiman,2, Nicole A. Doria-Rose 3, Constantinos Kurt Wibmer,2, Daniel J. Sheward 4, Carolyn Williamson 4,5, Salim S. Abdool Karim 5, Peter D. Kwong 3, John R. Mascola 3, Lynn Morris,2,5, Penny L. Moore,2,5 Centre for HIV and STIs, National Institute for Communicable Diseases of the National Health Laboratory Services, Johannesburg, South Africa, 2 School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 3 Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, South Africa, 4 Institute of Infectious Diseases and Molecular Medicine, Division of Medical Virology, University of Cape Town and NHLS, Cape Town, South Africa, 5 Centre for the AIDS Programme of Research in South Africa (CAPRISA), University of KwaZulu Natal, Durban, South Africa Background: A family of 2 anti-vv2 monoclonal antibodies (mabs) was previously isolated from an HIV- superinfected individual, CAP256. This lineage emerged weeks post-infection (w.p.i.) with individual mabs isolated at different time-points showing varying levels of maturation and neutralization activity. Mutations R66S or K69E became fixed at 76 w.p.i. and resulted in complete viral escape from all mabs. Here we investigated the effect of earlier mutations at positions 66 and 69 on the maturation of the CAP256-VRC26 lineage. Methods: Potential escape mutations were identified using single genome envelope amplification at time-points from 6-94 w.p.i. Mutations were introduced into the sensitive superinfecting virus and tested in neutralization assays against the 2 mabs to assess their role in escape. Results: By 42 w.p.i., just prior to the development of breadth, the wildtype 69K was replaced with either a 69I/R/Q/T in all sequences, indicating early immune pressure at this site. Introduction of K69I/Q mutations into the superinfecting virus abrogated neutralization by CAP256-VRC26.0 and 2, the least broad members of the family, with less effect on other lineage members. Similarly, at position 66, we observed a K66R mutation in 4/5 sequences at 94 w.p.i. that abrogated neutralization by all but the four broadest mabs, CAP256- VRC26.03, 04, 08 and 09. As 66R and 66K are the two most prevalent immunotypes at this position (in 70% and 4% of all global viruses, respectively), the increased breadth of these four mabs is consistent with their ability to tolerate both residues. Conclusions: Mutations that arose in the viral population soon after the expansion and subsequent maturation of the CAP256-VRC26 family exposed later lineage members to multiple immunotypes, thereby increasing their neutralization breadth. These data illustrate how a gradual process of autologous viral escape shaped the maturation of this mab response and provides insights for future sequential vaccine regimens. Elise Landais, Bryan S. Briney 2, Sergei L. Kosakovsky-Pond 3, Daniel T. MacLeod, Yolanda Lie 4, Paul Algate 5, Dennis R. Burton 2, Terri Wrin 4, Po-Ying Chan-Hui 5, Pascal Poignard,2, The IAVI Protocol C Investigators & The IAVI African HIV Research Network The International AIDS Vaccine Initiative, Neutralizing Antibody Center, La Jolla, CA, United States, 2 The Scripps Research Institute, Department of Immunology and Microbial Sciences, La Jolla, CA, United States, 3 University of California San Diego, Department of Medicine, La Jolla, CA, United States, 4 Monogram Biosciences Inc, San Francisco, CA, United States, 5 Theraclone Sciences Inc., Seattle, WA, United States Background: Designing a vaccine capable of eliciting broadly neutralizing antibodies (bnabs) to HIV requires a better understanding of these Ab maturation pathways as they occur by interplay with HIV envelope glycoprotein evolution. IAVI Protocol C is a large longitudinal cohort of primary HIV- infection in sub-saharan Africa. The development of bnab responses was evaluated in 385 Protocol C donors and individuals with bnab specificities targeting diverse highly conserved epitopes were selected for mab isolation and maturation studies. Methods: Donor PC64 serum neutralizing activity was mapped to a N60-glycan-dependent, kifunensine-sensitive VV2 quarternary epitope. The PG9-like neutralizing activity was detectable as early as 8 months post infection (mpi), peaked at 36 mpi and waned slightly during 2 years of additional follow-up. Envelope glycoprotein genes from PC64 were cloned at 0 different time points between and 48 mpi. High throughput B-cell activation and functional screening were used to isolate mabs with PG9-like neutralizing activity at 6 time points post infection. Deep sequencing of memory B-cells from longitudinal PBMCs samples was performed using the MiSeq (Illumina) platform. Results: Ten related mabs with PG9-like neutralizing activity were isolated from the month-36 sample. The Abs use the VH3-5*0 and VK3-20*0 genes and possess long CDRH3s (25 amino acids) that contain tyrosine residues. Deep sequencing of 2 longitudinal samples spanning 4 years of infection suggested the emergence of the lineage around 9 mpi, identifying a sequence with 99.66% nucleotide identity to germline. Analysis of Env sequences showed evidence of escape mutations and an increase in diversity after 2 mpi, likely a consequence of selection pressure from the PG9-like lineage. Conclusions: Additional Ab isolated from different time points are currently being characterized. The study will provide important insight regarding the maturation pathways of bnabs and critical information for vaccine design. 70 HIV Research for Prevention 204 HIV R4P

71 Wednesday, 29 October Oral Abstract Session 2: Towards Broadly Neutralizing Antibody Induction OA2.03 OA2.04 Development of Broadly Neutralizing Anti- HIV- Antibodies during Natural Infection through Early Epitope Acquisition and Subsequent Maturation D. Noah Sather, Sara Carbonetti, Delphine Malherbe 2, Franco Pissani 3, Andrew B. Stuart, Ann J. Hessell 2, Spyros Kalams 4, Nancy L. Haigwood 2, Leonidas Stamatatos Seattle Biomedical Research Institute, Seattle, WA, United States, 2 Oregon National Primate Research Center, Beaverton, OR, United States, 3 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 4 Vanderbilt University, Nashville, TN, United States Background: Approximately 20-30% of HIV- infected subjects develop broadly neutralizing antibodies (bnabs), which are prototypes for the types of antibodies that will likely need to be elicited by a successful HIV- vaccine. Delineating the key early events that lead to the development of bnabs during infection may help guide the development of immunogens and vaccine regimens to prevent HIV- infection. Methods: We followed two HIV- positive subjects from before they developed bnab activity until several years after breadth was detected in the plasma. We studies viral Envelope evolution over the course of infection and studied its relationship to the development of bnabs in early infection. Results: Both subjects developed bnab activity one year post infection, which ultimately mapped to the membrane proximal external region (MPER) and the CD4 receptor binding site. For one subject, we were able to identify anti-mper activity in the earliest plasma sample that exhibited no bnab activity, indicating that this epitope specificity was acquired very early on, but that these antibodies initially were not able to mediate neutralization. Escape mutations within the bnab epitopes did not arise in the circulating envelopes until bnab activity was detectable in the plasma, indicating that this early response was not sufficient to drive viral escape. Finally, we found that global anti-envelope binding avidity significantly increased at the time bnabs developed, indicating that antibody maturation was a key step. Conclusions: We report one potential mechanism by which bnabs develop during natural infection in which an epitope target is acquired very early on during the course of infection. After sufficient time and antibody maturation, the response matures to acquire broadly neutralizing activity, forcing the virus to escape. These findings reinforce a key role for antibody maturation in the development of bnabs, and highlight the necessity of developing vaccine regimens that are able to drive somatic hypermutation. Investigating Epitope Exposure on Native Trimers Sergey Menis, Chris Carrico 2, Dan Kulp, William Schief,3 The Scripps Research Institute, Department of Immunology and Microbial Sciences, La Jolla, CA, United States, 2 Buck Institute for Research on Aging, Novato, CA, United States, 3 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States Background: HIV infection typically generates autologous but not broadly neutralizing antibodies, presumably due to immunodominance of strain-specific epitopes on the native spike. Consistent with this view, it has been reported that rabbit immunization with the BG505 SOSIP soluble trimer, a faithful mimic of the native spike, induces potent BG505-neutralizing but not broadly neutralizing responses. We sought to employ the known structure of the BG505 SOSIP trimer to investigate the exposure of variable and conserved epitopes on the native spike. Methods: We computationally built a large ensemble of glycosylated BG505 SOSIP trimers with a variety of backbone and glycan conformations to approximate the motions of the native trimer. We then identified potential antibody epitopes in each member of the ensemble in an unbiased and exhaustive manner. Finally, we computed the frequency at which each position in the trimer is observed in an epitope. This frequency represents a quantitative estimate of the exposure to antibody for any particular position. Results: Antibodies with common CDR lengths access epitopes in the variable regions (V, V2, V4, V5) considerably more frequently (220 times more frequently) than in the conserved CD4-binding-site (CD4bs). Glycosylation is the primary culprit: in the absence of glycans, the variable regions are accessed only 7 times more frequently than the CD4bs. Dominant V5-directed responses may further reduce responses to conserved CD4bs epitopes, by competition. Increased CDRH3 length allows antibody access to hidden epitopes but also increases access to variable epitopes. Conclusions: Variable epitopes in V, V2, V4, V5 are predicted to be immunodominant on native spikes, purely on the basis of exposure. Antibodies against these epitopes likely inhibit induction of broadly neutralizing antibodies by trimer immunogens. Our analysis may guide design of trimers and immunization regimens for more favorable immune responses. ORAL ABSTRACT SESSIONS 7

72 Oral Abstract Sessions Oral Abstract Session 2: Towards Broadly Neutralizing Antibody Induction OA2.05 OA2.06 LB ORAL ABSTRACT SESSIONS An Inflammatory Profile that Predicts the Development of Neutralizing Antibody Breadth Anne Sophie Dugast, Kelly Arnold 2, Michelle Hoffner, Florencia Pereyra, Michael Seaman 3, Bruce Walker, Douglas Lauffenburger 2, Galit Alter Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 MIT, Department of Biological Engineering, Cambridge, MA, United States, 3 Center for Virology and Vaccine Research, Boston, MA, United States Background: Over the past decade, there has been an exponential increase in the discovery of new broadly neutralizing Abs (bnabs); however the mechanism by which these Abs are elicited remains unclear. Interestingly, compelling evidence from multiple studies suggests that high viremia and associated immune activation are potential drivers for the development of bnabs. Here we sought to dissect the inflammatory signals associated with bnab evolution in a large cohort of subjects that spontaneously control viral replication (Controllers) and to define whether viremia and inflammation can be unlinked as predictors of the induction of bnabs. Methods: Neutralizing activity and 9-plex cytokine analyses were performed in 03 Controllers and 7 acutely infected individuals who went on to develop bnabs versus 8 who did not. Acute subjects were followed longitudinally over 2 months. Partial least square regression and a decision tree analysis were used to define multivariate cytokine signatures associated with the evolution of bnabs across cohort groups. Results: We observed that high plasma levels of CXCL3, scd40l and IP0 were enriched in Controllers that evolved bnabs compared to those that did not (p=0.00). While none of the cytokines independently predicted bnab evolution, we found that a specific pattern of cytokine production was associated with the evolution of bnabs, consisting of high plasma levels of IP0 in combination with CXCL3 or scd40l. This profile was validated in a cohort of chronically infected patients that evolved bnabs, and predicted the evolution of neutralizing breadth in a cohort of acutely infected patients that went on to develop bnabs. Conclusions: Overall, this unique inflammatory signature points to a very specific mechanism for the evolution of neutralizing antibody breadth, that is induced as early as a few weeks post infection, pointing to a role of specific adjuvants or inflammatory cues that if co-delivered during immunization, may promote more effective affinity maturation. Induction of Antibodies with Long Variable Heavy Third Complementarity Determining Regions by Repetitive Boosting with AIDSVAX B/E in RV44 Vaccinees Michael Anthony Moody, David Easterhoff, Thaddeus C. Gurley, John F. Whitesides, Dawn J. Marshall, Andrew Foulger, Krissey E. Lloyd, Robert Parks, Justin Pollara, Ryan Duffy, Shaunna Shen, Jerome H. Kim 2, Nelson L. Michael 2, Merlin L. Robb 2, Robert J. O Connell 3, Sandhya Vasan 3, Jean- Louis Excler 2, Supachai Rerks-Ngarm 4, Jaranit Kaewkungwal 5, Punnee Pitisuttithum 5, Sorachai Nitayaphan 3, Faruk Sinangil 6, Donald Francis 6, Carter Lee 6, Thomas B. Kepler 7, S. Munir Alam, Guido Ferrari, David C. Montefiori, Hua-Xin Liao, Georgia D. Tomaras, Barton F. Haynes Duke University Medical Center, Duke Human Vaccine Institute, Durham, NC, United States, 2 US Military HIV Research Program, Rockville, MD, United States, 3 AFRIMS, Bangkok, Thailand, 4 Thai Ministry of Public Health, Nonthaburi, Thailand, 5 Mahidol University, Bangkok, Thailand, 6 Global Solutions for Infectious Diseases, South San Francisco, CA, United States, 7 Boston University, Boston, MA, United States Background: The Env gp20 AE.A244 used for boosting in the ALVAC/ AIDSVAX B/E RV44 trial expressed a dominant V2 linear epitope centered on lysine (K) at position 69, and a subdominant peptideglycan epitope recognized by VV2 broad neutralizing antibodies (bnabs) CH0 and PG9 and the unmutated ancestor antibody (Ab) of CH0. V2-specific Abs isolated from RV44 vaccinees displayed tier- strain-specific neutralization (AE.92TH023) and bound K69-containing linear epitopes. The RV305 trial recruited 90 RV44 participants to return after 6 years for two boosts with either ALVAC+AIDSVAX B/E or AIDSVAX B/E alone to evaluate the effect of this boost on vaccineinduced immunity. Methods: B-cell repertoires of 4 RV305 vaccinees with the greatest breadth of serum neutralization (A3R5 assay) were studied by antigenspecific memory B-cell sorting and recombinant Ab generation. Two vaccinees received ALVAC+AIDSVAX B/E and two were boosted with AIDSVAX B/E protein in alum. PCR-amplified monoclonal Abs (mabs) were characterized in binding, neutralization, and ADCC assays. Results: The boosts increased V H mutation frequency from that seen following the initial RV44 vaccine regimen (RV305 mean 5.50%, 258 mabs, 4 vaccinees; RV44 mean 2.60%, 05 mabs, 2 vaccinees) and expanded a population of Abs with heavy third complementarity determining regions (HCDR3s) > 22 amino acids (RV305 30/258; RV44 /05). Similar to VV2 bnabs- and other neutralizing Abs with long HCDR3s, these mabs principally used D2/D3 and J H 6. Moreover, 35.2% of mabs were sensitive to PNGase F native deglycosylation of Env gp20 AE.A244, including 9 mabs with long HCDR3s. Four vaccineinduced N56QN60Q-sensitive V2 mabs were isolated that are being characterized for neutralization capacity. Conclusions: Repetitive boosting of RV44 vaccinees expanded a pool of Abs with many of the characteristics of VV2 bnabs. Expansion of this subdominant group of antibodies by vaccination may represent a step forward in the quest to induce bnabs. 72 HIV Research for Prevention 204 HIV R4P

73 Wednesday, 29 October Oral Abstract Session 3: ARV Exposure & Efficacy in the Genital Tract OA3.0 OA3.02 Defining Pharmacokinetic and Pharmacodynamic linkages between Genital Tissue and Lumen Compartments Charlene S. Dezzutti,2, Lisa C. Rohan,2, Katherine Bunge, Nathan Ehrilich 2, Leslie Meyn 2, Philip Graebing 2, Mark Marzinke 3, Craig Hendrix 3, Brid Devlin 4, Sharon L. Hillier,2 University of Pittsburgh, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 3 Johns Hopkins University, Baltimore, MD, United States, 4 IPM, Silver Springs, MD, United States Background: Early clinical trials of topical microbicides only defined drug levels in tissue and luminal fluids. We are now determining if the drug quantity is sufficient to inhibit HIV. We theorize increased drug levels will correlate to decreased HIV infection. Methods: In the FAME-02 clinical trial, 60 women were randomized to use dapivirine (DPV) gel or film, placebo or active. After 7 daily doses of product, a 0 ml cervicovaginal lavage (CVL) and two vaginal biopsies were taken. For pharmacokinetic (PK) determination, DPV was quantified using LC-MS/MS. For pharmacodynamic (PD) activity, CVL was tested using an in vitro TZM-bl assay and tissue was tested using the ex vivo challenge assay. PK/PD relationships were determined by linear regression. Results: Women using the films and gels had no serious adverse events and found them to be acceptable. However, 5 women in the DPV film arm had not placed the films correctly and were excluded from this analysis. DPV levels in CVL ranged from ng/ml in film compared to ng/ml in gel users. DPV levels in vaginal tissue ranged from ng/mg in film compared to ng/mg in gel users. CVL showed a strong correlation between amount of DPV and inhibition of HIV infection from film (P < 0.000; r 2 = 0.635) and gel (P < 0.000; r 2 = 0.684) users. Likewise, vaginal biopsies showed a strong correlation between the DPV levels and inhibition of HIV infection in the tissue from film (P = 0.002; r 2 = 0.349) and gel (P < 0.000; r 2 = 0.530) users. For both CVL and tissue, there was no significant difference in drug levels or activity between film or gel users. Conclusions: To define the potency of a topical microbicide, PK and PD relationships are being defined. CVL had 0-00-fold higher DPV levels than vaginal tissue which was reflected in better PK/PD correlations in CVL as compared to tissue. While luminal concentrations were higher, these data show DPV was effectively delivered to both lumen and tissue compartments. Phase Safety & Pharmacokinetic Trial of a Polyurethane Tenofovir Disoproxil Fumarate Intravaginal Ring in Healthy, Low-risk U.S. Women Marla J. Keller, Lilia Espinoza, Mark A. Marzinke 2, Pedro M. Mesquita, Ashley M. Huber, Lorna K. Rabe 3, Bruce Frank 4, Jason McConnell 4, Mark Mitchnick 4, Craig W. Hendrix 2, Patrick F. Kiser 5, Betsy C. Herold Albert Einstein College of Medicine, Bronx, NY, United States, 2 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 3 Magee-Womens Research Institute, Pittsburgh, PA, United States, 4 Particle Sciences Inc., Bethlehem, PA, United States, 5 Northwestern University, Evanston, IL, United States Background: The prodrug tenofovir disoproxil fumarate (TDF) is more potent than tenofovir (TFV) against HIV in vitro & provides greater protection against genital herpes in a murine model. Sustained TDF ring delivery for 28 days (d) completely protected macaques from multiple vaginal simian-hiv challenges. Protection was associated with mean TFV levels in vaginal fluid (VF) of.8 x 0 5 ng/ml. The objectives of this trial are to evaluate the safety & pharmacokinetics of a reservoir-type TDF & placebo polyurethane (PU) IVR when used by women continuously for 4 d. Methods: A randomized, single blind, placebo controlled trial of 30 women is ongoing in Bronx, NY. Clinical safety is assessed by pelvic exam & colposcopy. Swabs (2 proximal, distal to IVR) and plasma are collected, 3, 7 & 4 d after ring insertion & 2 & 7 d after ring removal for drug levels. Cervical tissue is obtained for TFV & TFV-diphosphate levels & ex vivo HIV challenge studies. Anti-HIV & anti-hsv-2 activity of VF as a marker of pharmacodynamics is ongoing. Results: To date, women completed the study (5 TDF, 6 placebo). The mean age is 3 years (range 25-37). There are 4 product related Grade adverse events due to vaginal discharge (3 TDF, placebo) & no product related colposcopic findings. No subject reported ring removal or expulsions. Bacteria were not detected by culture of fluid from the core of 9 IVRs tested. Median VF TFV levels at, 3, 7 & 4 d after IVR insertion are 3, 4, 6 & 9 x 0 4 ng/ml, respectively. There was no significant difference in VF TFV levels during ring use but levels declined significantly 2 & 7 d after removal to 3 & 0.4 x 0 3 ng/ml, respectively (p=0.002). Conclusions: TDF & placebo PU vaginal rings are well tolerated & TFV levels exceed the clinical correlate of protection previously observed with TFV gel dosing before & after sex in women (~0 3 ng/ml). Sustained TDF IVR delivery may achieve drug concentrations needed to prevent HIV & herpes & overcome the low adherence seen in prior microbicide trials. ORAL ABSTRACT SESSIONS 73

74 Oral Abstract Sessions Oral Abstract Session 3: ARV Exposure & Efficacy in the Genital Tract OA3.03 OA3.04 ORAL ABSTRACT SESSIONS Multicompartmental Pharmacokinetics of Tenofovir % Gel Using the BAT 24 Regimen Versus Daily and Single Pericoital Dosing Jill L. Schwartz, Debra Weiner 2, Angela Kashuba 3, David Archer, Vivian Brache 4, Courtney A. Schreiber 5, Beatrice A. Chen 6, Alfred Poindexter 7, Andrea Thurman, Jaim J. Lai 2, Kuo H. Yang 3, Craig Sykes 3, Christine Mauck, Betsy Herold 8, Charlene Dezzutti 6, Gustavo F. Doncel CONRAD/EVMS, Arlington, VA, United States, 2 FHI-360, Durham, NC, United States, 3 University of North Carolina, Chapel Hill, NC, United States, 4 Profamilia, Santo Domingo, Dominican Republic, 5 Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States, 6 University of Pittsburgh, Pittsburgh, PA, United States, 7 Advances in Health, Houston, TX, United States, 8 Albert Einstein College of Medicine, Bronx, NY, United States Background: This multicenter study evaluated 90 women for multicompartment concentrations of tenofovir (TFV) and its active metabolite (TFV diphosphate [TFV-DP]) after vaginal administration of TFV % gel regimens (BAT24, pericoital and daily). Methods: TFV concentrations were measured in blood plasma, cervicovaginal fluid (CVF), & tissue, and TFV-DP was measured in tissue, using LC-MS/MS, after a single study-related sex act (SA) and after two weeks of twice weekly study-related sex (MA). For the SA, participants were randomized to BAT24, precoital or postcoital and to sample collection at 4, 2 or 72h after study-related sex. For the MA, they were re-randomized to BAT24, precoital, postcoital or daily (sample collection at 4, 2 or 72h and again at 5, 0 or 4 days after last study-related sex). Anti-HIV activity was measured in cervicovaginal lavages (CVL) by single-round infection assay. Results: Plasma TFV median concentrations were low ( 8ng/mL). Generally BAT24 produced the highest TFV in CVF and tissue, and the highest TFV-DP in tissue. Population composite median profiles of TFV and TFV-DP exposure in the tissues of BAT24 were -2x and 2-5x higher, respectively, compared to other arms. All regimens provided mean TFV- DP 800 fmol/mg, which persisted for up to 72 hours. The terminal half-lives of TFV and TFV-DP in the tissues were approximately 47h and 58h, respectively. Mean % HIV inhibition of CVL was 95% at 4h and 82% at 2h, but decreased to 52% by 72h, pooling all regimens. Conclusions: Although BAT24 generally produced higher TFV and TFV- DP, all dosing regimens yielded mean TFV-DP concentrations that were above a threshold associated with protection in nonhuman primates (~0 3 fmol). Decrease in % HIV inhibition in CVL at 72h was likely due to dilution. Persistently high TFV-DP concentrations in cervicovaginal HIV target tissues provide the basis for a forgiving dosing regimen. Based on these concentrations, the potential of a single pericoital dose should be explored. Population Pharmacokinetic Model of Vaginal Tenofovir % Gel in the Cervicovaginal Fluid Kuo-Hsiung Yang, Jill L. Schwartz 2, Craig Sykes, Gustavo F. Doncel 2, Angela D.M. Kashuba UNC Eshelman School of Pharmacy, Chapel Hill, NC, United States, 2 CONRAD/EVMS, Arlington, VA, United States Background: Tenofovir (TFV) gel effectiveness trials have measured TFV concentrations in cervicovaginal fluid (CVF) from trial participants to determine product adherence and to assess the relationship between adherence and effectiveness. Herein we present a model which describes TFV gel CVF pharmacokinetics (PK). Methods: We evaluated data (N=) from 2 PK studies. TFV concentrations were measured in CVF after a single dose (CONRAD095), study-related sex act & 2 weeks of twice weekly study-related sex acts (CONRAD 3). Nonlinear mixed effects population PK modeling was performed with NONMEM 7.2. Inter-individual variability (random effects) was evaluated with an exponential error model while residual variability was assessed with additive & proportional error model. 3 Monte-Carlo Simulation runs were performed to generate prediction intervals for BAT24, peri-coital, and daily dosing (stead-state) regimens. Results: A 3 compartment, bolus input, linear kinetic model with a vaginal gel depot compartment, st-order absorption (Ka) into CVF (Vb), clearing into Vc with distributional clearance (CLd), and st-order elimination (CLt) best described the data. Fixed effects for Ka, CLt, CLd, Vb, Vc were:.54 (/h), 9.56 & 0.04 x 0-4 (L/h), 06 & 2.56 x 0-4 L. The inter-individual variability (CV%) for the parameters were: 23, 82, 22, 45, and 28 with a full covariance matrix on CLt, CLd, Vc, and Vb (residual additive error: 0.65; proportional: 0.32). Monte-Carlo Simulations from the model predicted that 95% of the population would have CVF TFV concentrations falling below 2ng/mL (LLQ) by week 6 after last gel dose for peri-coital & BAT24 regimens, and week 7 for daily dosing(median terminal T /2 = 43h). Conclusions: This model highlights the very long cervicovaginal decay of TFV gel after single or multiple applications. Furthermore, it can be used to predict CVF TFV concentrations, and is therefore useful for objective assessment of clinical trial adherence & product development benchmarks. 74 HIV Research for Prevention 204 HIV R4P

75 Wednesday, 29 October Oral Abstract Session 3: ARV Exposure & Efficacy in the Genital Tract OA3.05 LB OA3.06 Sex Matters: MTN-0 Phase Study on the Impact of Sex on Tenofovir Gel Pharmacokinetics (PK) and Pharmacodynamics (PD) Betsy C. Herold, Cliff Kelly 2, Beatrice Chen 3, Robert Salata 4, Mark Marzinke 5, Charlene Dezzutti 3, Lisa Levy 6, Beth Galaska Burzuk 7, Jeanna Piper 8, Ian McGowan 7, Craig W. Hendrix 5, MTN 0 Albert Einstein College of Medicine, Bronx, NY, United States, 2 SCHARP (Statistical Center for HIV/AIDS Research and Prevention) Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 3 Magee Womens Research Institute, Pittsburgh, PA, United States, 4 Case Western Reserve University, School of Dental Medicine, Cleveland, OH, United States, 5 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 6 Family Health International, Washington, DC, United States, 7 Magee Women Research Institute, Pittsburgh, PA, United States, 8 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States Background: Tenofovir (TFV) gel reduced HIV acquisition when applied pericoitally in CAPRISA 004, but not with daily dosing in VOICE, attributed largely to poor adherence. The timing of gel dosing relative to sex also may have contributed to trial outcomes as sex/semen may modify local TFV concentration. To identify optimal gel dose timing, MTN conducted a phase PK/PD study of TFV % gel applied hour before (-h), 24 hours before (-24h), or hour before and hour after (BAT) sex compared to dosing without coitus. Methods: The trial enrolled 24 evaluable couples from 2 US sites. Women completed 2 baseline no gel visits (no sex, sex) and 5 gel visits (- h + sex, - h no sex, -24 h + sex, -24 h no sex, and BAT + sex). Cervicovaginal lavage (CVL), vaginal and cervical tissues, and plasma were collected ~2 h after sex with matching collection times at no sex visits and assayed for drug (PK) and CVL anti-hiv activity in vitro (PD). Results: BAT dosing achieved the highest (CVL: ng/ml; cervical: 29 ng/mg; vaginal: 258 ng/mg) and -24 h + sex the lowest (CVL: ng/ml; cervical:.46 ng/mg; vaginal: 5.3 ng/mg) TFV levels. Compared to dosing without sex, mean TFV levels after sex decreased 42% and 78% (.33x0 5 ng/ml, p=0.005 and 8.53x0 3 ng/ml, p< 0.00) in CVL and decreased 74% and 55% (3.92 ng/mg, p=0.04 and 2.64 ng/mg, p< 0.00) in cervical tissue with - h and -24 h dosing, respectively. Vaginal tissue decreases were even greater. In contrast, mean plasma TFV was 28% higher (.6 ng/ml, p< 0.0) following sex with - h dosing, presumably reflecting greater absorption. Postcoital CVL anti-hiv activity increased significantly from a median [IQR] baseline of 55[54]% in the absence of gel to 99[7], 77[57], and 00[0.4] with -, -24, or BAT dosing, respectively. The antiviral activity of CVL correlated significantly with drug level. Conclusions: Timing of dosing relative to sex impacts TFV gel PK/ PD. Pericoital dosing or sustained delivery may be optimal for PrEP, particularly with poor adherence. Population Pharmacokinetic Modelling of Dapivirine Neliette Van Niekerk, Jeremy Nuttall 2, Stefan Zeiser 3, Annalene Nel International Partnership for Microbicides, Clinical Affairs, Paarl, South Africa, 2 International Partnership for Microbicides, Product Development, Silver Spring, MD, United States, 3 Kinesis Pharma BV, PK/PD Modelling, Breda, Netherlands Background: A population pharmacokinetic (PK) model has been developed for dapivirine delivered from the Dapivirine Vaginal Ring-004 (25 mg dapivirine). The model describes the uptake, distribution and elimination of dapivirine, and allows the simulation of scenarios that have not been tested, allowing informed decision-making without the need for additional clinical trials. Methods: The population PK model was developed to describe dapivirine levels in vaginal fluid (CVF) at the cervix and introïtus, as well as in plasma, based on data from two Phase I trials, IPM 03 and IPM 024, which evaluated the local and systemic PK of dapivirine from Ring- 004 in healthy, HIV-negative women. The model was validated with results from the dapivirine-only arm in IPM 028, a drug-drug interaction study with miconazole nitrate. Results: Different types of model structures were evaluated. The observed dapivirine plasma and CVF concentrations were best described using a model with direct transition from the fluid to the plasma compartment, including a peripheral compartment. Visual predictive checks were good, and fixed and random effects parameters could be estimated with high precision. Although the initial purpose of the model was to predict dapivirine CVF levels in cases of ring loss/removal, it also proved very useful in assessing non-adherence to ring use in the ongoing IPM 027 Phase III trial. The effect of different ring use/removal scenarios on dapivirine plasma and CVF concentrations was simulated and compared to expected levels associated with perfect ring use over 28 days. This assisted in defining a range of dapivirine plasma and CVF levels that may be indicative of non-adherence to ring use over 28 days. Conclusions: The population PK model for dapivirine allows the simulation of ring use scenarios not tested in clinical trials (effect on drug levels of ring removal, prediction of accumulation for successive ring insertions), and is a very useful tool to support non-adherence evaluations in Phase III. ORAL ABSTRACT SESSIONS 75

76 Oral Abstract Sessions Oral Abstract Session 4: Host Factors: Injury, Acquisition and Infection OA4.0 OA4.02 ORAL ABSTRACT SESSIONS Protective HLA Alleles Reduce Markers of Gut Damage and Microbial Translocation and Preserve the Cellular Immune Response during Acute HIV- Infection Daniel Claiborne, Jessica Prince, Eileen Scully 2, Jakob Kopycinski 3, Gladys Macharia 3, Krystelle Nganou-Makamdop 4, Jianming Tang 5, Paul Goepfert 5, Tianwei Yu, Shabir Lakhi 6, William Kilembe 6, Daniel Douek 4, Jill Gilmour 3, Matthew Price 3,7, Marcus Altfeld 8, Susan Allen,6, Eric Hunter Emory University, Atlanta, GA, United States, 2 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States, 3 IAVI, London, United Kingdom, 4 NIAID, NIH, Bethesda, MD, United States, 5 University of Alabama at Birmingham, Birmingham, AL, United States, 6 ZEHRP, Lusaka, Zambia, 7 UCSF, Epidemiology & Biostatistics, San Francisco, CA, United States, 8 Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany Background: Efforts to elucidate protective host factors in HIV- infection often rely on cross-sectional data to determine factors influencing pathogenesis. Here, we study a cohort of 27 acutely infected Zambians with longitudinal CD4 counts up to 5 years post infection to identify novel HLA alleles associated with disease progression. Methods: Cox proportional hazard models were used to elucidate protective and deleterious HLA alleles. Plasma cytokines were measured at seroconversion (median 45 days post infection), 3, and 6 months post infection using the Luminex platform. Multicolor flow cytometry was used to assess cellular activation. Results: In a multivariable Cox model, HLA-B*40, B*57, B*580, B*8, DQB*02, and DRB*5 were found to provide significant protection from CD4 T cell decline. Protective HLA class I alleles were associated with significantly lower plasma lipopolysaccharide (LPS) levels and fewer activated (CD38+, PD-+) CD8+ T cells early in acute infection, a time point before these protective alleles significantly altered plasma viral load (VL). Furthermore, protective HLA class I alleles were associated with a decrease in IL-0 levels over time and lower levels of ifabp in plasma at 6 months post infection. This data suggests that protective cellular immune responses operate early in acute infection before control of plasma VL is detected. Conclusions: This study of a well-characterized subtype C Zambian cohort of acutely infected individuals provides an opportunity to elucidate host immunogenetic factors contributing to HIV- pathogenesis and to investigate the underlying immunological mechanisms. This data suggests that protective HLA alleles can influence disease course early in acute infection by maintaining gut integrity, limiting microbial translocation, and ultimately preserving functional cellular immune responses by reducing inflammation. Combined Effect of HLA-C*04:0 and KIR2DS4 on Increased HIV Viral Loads Alex Olvera, Susana Pérez-Alvarez,2, Carmina Ganoza 3, Javier Lama 3, Nicole Bernard 4, Jorge Sanchez 3, Christian Brander,2,5 IrsiCaixa AIDS Research Institute - HIVACAT, Badalona, Spain, 2 Universitat Autònoma de Barcelona, Barcelona, Spain, 3 Asociación Civil IMPACTA Salud y Educacion, Lima, Peru, 4 Research Institute of the McGill University Health Centre, Montreal, QC, Canada, 5 Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain Background: It is well known that relative viral control can be achieved by some HIV infected individuals, especially those expressing particular HLA class I and KIR genes. Since KIR genes use HLA alleles as ligands, HLA and KIR polymorphisms may both be related to relative HIV control. However, specific HLA-KIR binding combinations are largely unknown and the effect of these combinations on HIV viral control is just beginning to emerge. Methods: We assessed potential associations between HLA alleles and KIR genes with HIV viral load and CD4 counts in a cohort of 235 untreated individuals with HIV clade B infection in Lima (Peru). All individuals were HLA typed at a 4-digit resolution, 70 of them were also typed for KIR genes. For the KIR2DS4 locus, full (f) and deleted (d) alleles were differentiated. Differences in viral load and CD4 counts were tested by Mann-Whitney test and multiple comparisons effects were controlled applying False Discovery Rate (FDR) and q-value calculation. Results: Nineteen HLA alleles and 3 KIR genes were associated with differences in viral load and/or CD4 counts (based on uncorrected p < 0.05). Of these, only the association between HLA-C*04:0 with high viral load remained statistically significant after multiple comparison correction (p=0.000, q=0.0097). Interestingly, the 3 KIR genes showing differences with p< 0.05 were all putative HLA-C*04:0 ligands. For this reason we evaluated the combined effect of C*04:0 with the KIR2L, KIR2DS and KIR2DS4. Our results show that the association between HLA-C*04:0 and elevated viral load was due to the co-expression of KIR2DS4f, but not KIR2DS4d. Conclusions: Our data indicate that the association between HLA-C*04:0 expression and high viral load and reduced CD4 count is due to a NK cell component that has not been appreciated before. This observation, together with other reports on HLA-KIR allele combination and variable HIV control may help shed further light on immune mechanisms of HIV control and guide HIV immunogen design. 76 HIV Research for Prevention 204 HIV R4P

77 Wednesday, 29 October Oral Abstract Session 4: Host Factors: Injury, Acquisition and Infection OA4.03 Immune Correlates Identified in the RV44 Vaccine Efficacy Trial Impact HIV- Acquisition Only in the Presence of Certain HLA Class II Genes OA4.04 LB Transmitted Escape Mutations Lead to Accelerated HIV- Disease Progression and Largely Define the Relative Contribution of HLA Alleles to Control Heather Prentice,2, Daniel Geraghty 3, Georgia D. Tomaras 4, Youyi Fong 5, Wyatt Nelson 3, Gustavo Kijak,2, Susan Zolla- Pazner 6, Sorachai Nitayaphan 7, Supachai Rerks-Ngarm 8, Jaranit Kaewkungwal 9, Punnee Pitisuttithum 9, Nelson L. Michael, Peter Gilbert 5, Jerome H. Kim, Rasmi Thomas,2 U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Silver Spring, MD, United States, 2 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 3 Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 Duke University Human Vaccine Institute and the Center for HIV/AIDS Vaccine Immunology, Duke University School of Medicine, Durham, NC, United States, 5 Statistical Center for HIV/AIDS Research and Prevention, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 6 Veterans Affairs New York Harbor Healthcare System and the Department of Pathology, New York University School of Medicine, New York, MD, United States, 7 Department of Retrovirology, US Army Medical Component, AFRIMS, Bangkok, Thailand, 8 Department of Disease Control, Ministry of Public Health, Nonthaburi, Thailand, 9 Center of Excellence for Biomedical and Public Health Informatics BIOPHICS, Mahidol University, Bangkok, Thailand Background: Two immune correlates were identified as predictors of risk of infection in the RV44 HIV- vaccine clinical trial. Binding of plasma IgA antibodies (IgA) to HIV- envelope (Env) correlated directly with acquisition whereas binding of IgG antibodies to an antigen containing the variable regions and 2 of Env (IgG) correlated inversely with acquisition. We hypothesized that HLA class II molecules expressed on antigen presenting cells modulate CD4 T cell stimulation of antibody production by B cells involved in vaccine-induced responses. Methods: HLA class II genes were genotyped in the case-control group of 205 uninfected and 4 infected RV44 vaccinated volunteers. The interaction of HLA-DPB, DQB and DRB alleles with IgA and IgG immune responses was tested for an effect on acquisition by logistic regression. Results: DQB*06 (p=0.002, q=0.06), DPB*05 (p=0.03, q=0.9), and DPB*3 (p=0.04, q=0.20) had a significant interaction with IgA antibody responses on acquisition. Increased IgA levels associated with increased risk for infection only in the presence of DQB*06. In contrast, IgA levels did not associate with increased risk for infection in the presence of DPB*05 and *3. Further, high IgG antibody levels directly correlated with the presence of DPB*3 allele. IgG VV2 antibodies were associated with reduced acquisition risk only in the presence of DPB*3 (OR=0.3, p=0.007) with no association observed in the absence of DPB*3. The observed interactions for DQB*06 with IgA and DPB*3 with IgG remained significant in a multivariable model when testing other primary variables including IgG avidity, antibodydependent cellular cytotoxicity, neutralizing antibodies, and Env-specific CD4+ T cells. Conclusions: These associations suggest that certain HLA class II genes modulated the effect of antibody responses to the RV44 vaccine and impacted HIV- acquisition. Understanding this HLA class II mechanism will enable improved HIV vaccine design. Jonathan M. Carlson, Malinda Schaefer 2, Chanson Brumme 3, Nico Pfeifer, Roger Shapiro 4, Thumbi Ndung u 5, John Frater 6, Simon Mallal 7, Mina John 7, David Heckerman, Philip Goulder 6, Zabrina Brumme 8, Eric Hunter 2 Microsoft Research, Redmond, WA, United States, 2 Emory Vaccine Center, Atlanta, GA, United States, 3 UBC Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada, 4 Beth Deaconess Medical Center, Cambridge, MA, United States, 5 University of KwaZulu Natal, Durban, South Africa, 6 University of Oxford, Oxford, United Kingdom, 7 Murdoch University, Murdoch, Australia, 8 Simon Fraser University, Burnaby, BC, Canada Background: The extent of intra- and inter-host adaptation of HIV to HLA-mediated immune responses remains a central challenge for vaccine design, as the presence of escape mutations in circulating HIV sequences may compromise vaccine-induced immunity and reduce the protective effects of certain HLA alleles. One strategy is to vaccinate against escaped epitopes, though it is unclear whether such epitopes can elicit effective immune responses. Methods: We developed a probabilistic model of HIV sequence evolution, trained on >4,000 clade B and C sequences with matched HLA types, that yields a natural metric of the extent of HLA-specific adaptation of each sequence. HIV adaptation to HLA was strongly associated with VL and CD4 counts in cross-sectional and longitudinal early infection cohorts, validating our models and confirming the role autologous adaptation plays in disease progression. Results: Within transmission pairs, the extent to which the donors Gag, Pol and Nef sequences were pre-adapted to the linked recipients HLA alleles predicted recipient VL 24 mo post infection (Spearman r =0.39, p=0.0005, N=8) and time to CD4< 250 (HR = 5.97, p = 0.03, N=48). In cross-sectional chronic cohorts, individuals for whom the regional circulating viral sequences were well adapted had higher viral loads. Furthermore, the extent to which circulating HIV strains were adapted to individual HLA alleles predicted the VL associated with those alleles (Spearman r=0.77, p=0.009). Conclusions: Transmission of HIV pre-adapted to host HLA leads to accelerated disease progression and largely explains the extent to which an HLA is relatively protective or hazardous. This observation argues that the immune system is unable to mount effective responses against these adapted epitopes, even when they represent the initial challenge variant, and suggests that vaccination strategies that target such adapted epitopes must achieve a level of response that is not observed in natural infection. ORAL ABSTRACT SESSIONS 77

78 Oral Abstract Sessions Oral Abstract Session 4: Host Factors: Injury, Acquisition and Infection OA4.05 OA4.06 ORAL ABSTRACT SESSIONS Inhibiting the Host Exonuclease TREX Induces a Localized and Protective Host Interferon Response against Acute HIV Infection in vivo Lee Adam Wheeler, Judy Lieberman Harvard Medical School, Boston, MA, United States Background: Viral infections typically trigger innate immune sensors to produce type I interferons (IFNs). HIV infection however, does not activate these pathways. We previously showed that by inhibiting the cytosolic exonuclease TREX in vitro, HIV DNA accumulated in the cytoplasm and up-regulated IFNs that reduced HIV replication and subsequent spreading. Methods: Here, we use RNA interference to investigate the effect of knocking down trex expression in CD4+ cells on HIV transmission in human cervicovaginal explants, and in vivo, using a small animal model of HIV. Results: In the absence of TREX, IFNs and interferon response elements (IREs) are induced in vaginal tissue exposed to HIV and effectively suppress viral replication, thereby blocking HIV transmission to human cervicovaginal tissue and delayed transmission to humanized mice for ~3-4 weeks relative to mock-treated controls. Protection from HIV transmission was coupled with a marked increase in type I IFN expression. Expression of inflammatory cytokines (IL-b, IL-6, IL- 8), which facilitate HIV transmission, is significantly reduced relative to mock- and LPS-treated controls. Direct treatment of vaginal tissue with recombinant IFNβ also decreased viral replication (albeit less effectively than inhibiting trex), though it did not block the induction of inflammatory cytokines. This suggests that the accumulation of viral DNA in the cytosol of infected cells, which TREX is known to degrade, is an important trigger to the induction of these genes both in human tissues and in vivo. Conclusions: From these data, we conclude that, in the absence of TREX expression, the host interferon response can effectively control viral replication and block dissemination for several weeks. Further characterization of TREX function in vivo will allow for a better understanding of the mechanism by which HIV evades the host antiviral response and could provide new targets for drug and/or vaccine development to prevent HIV transmission. Candidate Loci Associated with AIDS Virus Replication Identified by Whole Genome Sequencing of SIV-Infected Macaques Adam Ericsen, Gabriel Starrett, Justin Greene, Michael Lauck, Mariel Mohns, Brian Cain, Ngoc Pham, Muthuswamy Raveendran 2, David Rio Deiros 2, Jason Weinfurter 2, Adam Bailey, Julie Karl, Roger Wiseman, Donna Muzny 2, Thomas Friedrich, Richard Gibbs 2, Jeffrey Rogers 2, David H. O Connor 3 University of Wisconsin - Madison, Madison, WI, United States, 2 Baylor College of Medicine, Houston, TX, United States, 3 University of Wisconsin - Madison, Pathology and Laboratory Medicine, Madison, WI, United States Background: A small percentage of HIV-infected people and SIVinfected macaques spontaneously control chronic-phase viral replication. This control is associated with certain major histocompatibility complex (MHC) class I alleles. However, only a minority of individuals with these protective MHC class I alleles suppress viral replication, suggesting additional loci act in concert with MHC genetics to mediate viral control. Methods: We sequenced whole genomes of 2 SIV-infected Mauritian cynomolgus macaques (6 controllers and 6 progressors), all of whom share the control-associated MHC haplotype termed M, and identified genetic variation that differentiated controllers from progressors. We assembled a second cohort to prospectively test the role of one such region on SIV control. Results: We found the highest densities of control-associated variation on chromosomes 2, 3, 7, and 9. One of these regions is near (within ~400 kilobases) a cluster of genes involved in antigen presentation and CD8(+) effector cell function, including the gene encoding the cytotoxic granule protein granzyme B (GzmB). The inheritance of a single granzyme B allele was associated with chronic-phase control of SIVmac239 (P=0.000) and higher expression of GzmB by CD3(-) CD8(+) NK cells during early infection. We then assembled a cohort of MHC-identical M(+) animals to prospectively test whether M(+) animals with this protective GzmB allele control SIV infection; however, only of 4 animals in this cohort controlled SIV. Conclusions: Although we did not confirm a causal relationship between GzmB genetics and MHC-associated control of SIV, we established a framework for identifying candidate control-associated loci and prospectively testing causality. We are currently assessing whether any of these regions differentiate a larger cohort of Indian-origin rhesus macaque controllers (n=20) from the Indian-origin rhesus population in general (n=44). 78 HIV Research for Prevention 204 HIV R4P

79 Wednesday, 29 October Oral Abstract Session 5: PrEP and Microbicide Adherence in Women OA5.0 OA5.02 Hidden Heterogeneity: Uncovering Patterns of Adherence in Microbicide Trials Lori Miller, Richard Hayes London School of Hygiene & Tropical Medicine, London, United Kingdom Background: Understanding participant adherence is critical to interpret results from HIV prevention microbicide trials. Typically trials report microbicide adherence based on the average of the number of selfreported sex acts which are covered by gel during the trial. Past trial estimates indicate high adherence, often around 80-95%. In reality, the population of participants may be composed of subgroups of women who have different patterns of gel use over time. Thus, a longitudinal approach to analyse adherence data may provide a better understanding of how participants use study products over follow-up. This study sought to identify subgroups of women with different adherence trajectories over the course of several completed microbicide trials. Methods: Longitudinal latent class analysis (LLCA) was used to identify groups of participants with different adherence trajectories over trial follow-up. Self-reported use of gel at sex acts was analysed from four microbicide trials: MDP 30, HPTN 035, Cellulose Sulfate CONRAD, and Carraguard. LLCA models were selected for each trial based on best fit and interpretably, and compared across trials. Results: Preliminary results identified several subgroups of women with different adherence patterns over time, demonstrating that not all participants had consistently high adherence. The different groups included very high adherers throughout the trial, those with diminishing adherence over time, and those with mid to high adherence. Interestingly, groups with initially low adherence which increased over time were not identified. Conclusions: LLCA revealed that trials are actually composed of a heterogeneous mixture of adherence patterns among trial participants. LLCA provided a more nuanced understanding of self-reported data. Results from trajectory analysis can be used to identify participant characteristics associated with different adherence patterns, and this information may be helpful for recruitment or adherence counselling for future microbicide trials. To Give or Not to Give PK Results: An Ethical Dilemma for Researchers & Regulators in Uganda Juliane Etima, Clemensia Nakabiito, Carolyne A. Akello, Samuel Kabwigu, Teopista Nakyanzi, Josephine Nabukeera, Miriam Hartmann 2, Elizabeth Montgomery 2, Barbara Mensch 3, Ariane Van der Straten 2,4 Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 Women s Global Health Imperative, RTI International, San Francisco, CA, United States, 3 Population Council, New York, NY, United States, 4 Center for AIDS Prevention Studies (CAPS), UCSF, Dept. of Medicine, San Francisco, CA, United States Background: During the VOICE trial, self-reports and monthly product counts were used to measure adherence. Despite high reported adherence, drug pharmacokinetic (PK) levels indicated widespread nonuse. The second stage of MTN 003D, a qualitative ancillary study to VOICE, retrospectively provided participants with PK results in hopes of encouraging candid discussion of low adherence. As the first PrEP study to provide such results directly to participants, researchers and regulators in Uganda were challenged with several ethical concerns. Methods: After weighing risks and benefits of disclosing individual PK results during in depth interviews and prior to focus group discussions, tools were developed to ensure these were presented in a clear, yet neutral manner using pictures of a teapot with varying amounts of liquid. Nonetheless, regulators were concerned about the lack of health benefit and possible victimization including blame and/or ill treatment of participants for non-use. Study teams underwent training on the meaning of results, how to provide them in a non-judgmental manner; and how to handle participants reactions. Regulators concerns were allayed during a face-to-face meeting with members of the study team who described how results would be provided and how participants, who might be upset by the results, would be counseled. Results: In Uganda, 5participants received PK results; reactions ranged from disbelief to joy. No social harms were noted. Use of the tools facilitated discussions, even after initial denial of results. Indeed, while only 3/4 low adherers initially accepted their results, during the course of the interview many went on to provide reasons for non-use. Conclusions: Despite ethical concerns, provision of PK results can be done in a safe and ethical manner, which reduces the possibility of victimization. The provision of these results can provide useful insights into participants adherence challenges and future design of trials. ORAL ABSTRACT SESSIONS 79

80 Oral Abstract Sessions Oral Abstract Session 5: PrEP and Microbicide Adherence in Women OA5.03 OA5.04 ORAL ABSTRACT SESSIONS The Effect of Presentation of Pharmacokinetic (PK) Drug Results on Self-reported Study Product Adherence among VOICE Participants in Zimbabwe Petina Musara, Otillia Munaiwa, Imelda Mahaka 2, Nyaradzo M. Mgodi, Miriam Hartmann 3, Lisa Levy 4, Elizabeth T. Montgomery 3, Cynthia I. Grossmann 5, Zvavahera Mike Chirenje, Ariane van der Straten 3, Barbara Mensch 6 University of Zimbabwe-University of California San Francisco Collaborative Research Programme, Harare, Zimbabwe, 2 Zimbabwe AIDS Prevention Project, University of Zimbabwe, Harare, Zimbabwe, 3 Women s Global Health Imperative; RTI International, San Francisco, CA, United States, 4 FHI-360, Durham, NC, United States, 5 DAIDS/NIH/ NIMH, Bethesda, MD, United States, 6 Population Council, New York, NY, United States Background: Honest reporting of participant product use is critical to understanding challenges to adherence in HIV PrEP trials. Previous studies, like VOICE, have shown a huge gap between self reported adherence and actual PK data. MTN-003D study was conducted after VOICE trial to elicit truthful reports about product non-use in VOICE. The study was conducted in 2 stages, the first without PK drug results provided to participants, the second with. Methods: We conducted in-depth interviews and focus group discussions. Guides included questions assessing factors affecting women s use of study product. We enrolled 74 women at 2 Zimbabwe sites (26 in stage ; 48 in stage 2.) Nine participated in both stages, and constitute the sample for this analysis. We analyzed changes in responses to adherence questions among the women in both stages to gain insight into their adherence challenges. Data from both stages were coded, summarised and compared using NVivo. Results: Five out of the 9 women reported high adherence in stage, yet admitted to non-use in stage 2. PK drug results showed that 2 were low adherers and 3 were inconsistent users. Zero of the 5 reported any major challenges to product use in stage, but rather described facilitators such as supportive sexual partner / family, perceived high HIV risk and regular use of reminders. In stage 2, participants divulged several reasons for product non-use, including joining the study for health services, partner disapproval, side effects, decreased sexual pleasure, negative influence, perceived reduced HIV risk, and difficulty swallowing tablets. One participant reported high adherence in stage, which she maintained in stage 2, despite PK results to the contrary. Three participants had consistent responses in stages and 2. Their PK drug results indicated that they were high adherers. Conclusions: Presentation of drug PK results to study participants may facilitate more honest reporting of product use and challenges to adherence. Disclosure of Pharmacokinetic (PK) Drug Results Promotes Open Discourse on Nonadherence among Women in VOICE Ariane van der Straten,2, Petina Musara 3, Juliane Etima 4, Kubashni Woeber 5, Elizabeth T. Montgomery, Miriam Hartmann, Lisa Levy 6, Thola Benny 7, Helen Cheng, Jeanna Piper 8, Cynthia I. Grossman 9, Barbara Mensch 0, MTN-003D Study Team Women s Global Health Imperative; RTI International, San Francisco, CA, United States, 2 University of California San Francisco, CAPS, Department of Medicine, San Francisco, CA, United States, 3 UZ-UCSF Collaborative Research Programme, Harare, Zimbabwe, 4 Makerere University Johns Hopkins University Research Collaboration, Kampala, Uganda, 5 Medical Research Council, Durban, South Africa, 6 FHI 360, Durham, NC, United States, 7 Desmond Tutu HIV Foundation, Cape Town, South Africa, 8 DAIDS/NIAID/NIH, Bethesda, MD, United States, 9 NIMH/NIH, Bethesda, MD, United States, 0 Population Council, New York, NY, United States Background: In VOICE, 50% of women assigned to daily active oral tablets or vaginal gel had undetectable drug in all plasma samples tested. MTN-003D is an ancillary study using in-depth interviews (IDI) and focus group discussions (FGD), together with disclosure of PK results, to explore participants adherence challenges during VOICE. Methods: We systematically recruited VOICE participants, with plasma samples tested (median=6 samples), and PK results defined as low (0%, N=85), inconsistent (>0% to< 75%, N=26) or high ( 75%; N=20) based on frequency of drug detection. The PK results were disclosed one-on-one and initial reactions captured by interviewers. 72 IDI and 2 FGD were conducted and summarized in debriefing reports for rapid review of key findings. IDI participants also ranked 20 theme cards listing adherence challenges. Results: There were 3 participants (32 South Africa, 48 Zimbabwe, 5 Uganda). The most common reaction to disclosed PK results, by group was: surprise (40%; low), acceptance (42%; inconsistent) and happiness (65%; high). While the majority accepted their PK results in Zimbabwe and South Africa, the opposite happened in Uganda. Nevertheless, even those disagreeing with results discussed adherence challenges. Those in the low/inconsistent groups described how they disposed of unused products. Preliminary findings indicate that most reported initiating use; however, low adherers later discontinued or poorly executed product use. Experienced/worried about side effects and products may be harmful were among the 3 top ranked adherence challenges in both the low and high PK groups. Not receiving enough support was ranked third in the low group; but much lower ( th ) in the high group. Conclusions: Provision of PK results seemingly promoted candid discussions around poor adherence and experience with products in VOICE. Detailed analyses of transcripts should clarify the meaning behind site differences in PK results reactions, and adherence challenges reported by each PK groups. 80 HIV Research for Prevention 204 HIV R4P

81 Wednesday, 29 October Oral Abstract Session 5: PrEP and Microbicide Adherence in Women OA5.05 OA5.06 LB Bounding the Seasons of Pre-exposure Prophylaxis: When and why Women at Higher Risk of HIV Might Suspend Use Emily E. Namey, Amy Corneli, Kawango Agot 2, Khatija Ahmed 3, Jacob Odhiambo 2, Joseph Skhosana 3 FHI 360, Social & Behavioral Health Sciences, Durham, NC, United States, 2 Impact Research and Development Organization, Kisumu, Kenya, 3 Setshaba Research Centre, Pretoria, South Africa Background: Now that Truvada as pre-exposure prophylaxis (PrEP) has been shown to be effective, critical questions about implementation center on when individuals should take PrEP and for how long. Periods of PrEP use, suggested to correspond with periods of increased HIV risk, have been termed seasons of PrEP. Understanding when and why women would suspend PrEP use is essential for effective counseling on initiating and stopping PrEP. We explored how African women at higher risk of HIV might define the end of a season of PrEP, to help inform counseling guidelines. Methods: As part of a larger study exploring PrEP and risk compensation among women at higher risk of HIV in Bondo, Kenya, and Soshanguve, South Africa, we conducted qualitative semi-structured interviews with 30 women from each site. Participants created timelines of their sexual contacts over the past six months and were asked to imagine they had taken PrEP during this period. Referencing their timelines, the women discussed whether and why they would have taken a break from PrEP, or would do so in the future. Results: A third of the women reported they would have suspended their use of PrEP during the past six months. Nearly half foresaw a reason to suspend PrEP use in the future, citing reasons related to partnership issues, phases of life, and life events. Partnership issues ranged from no current partner to concepts of trust and stability in relationships. Phases of life included pregnancy and aging out of frequent sex. Life events focused on extended travel or festive seasons that could disrupt PrEP use. In some of these contexts, suspension of PrEP was related to a perceived reduction in HIV risk. In others, the potential for risk stayed the same or increased. Conclusions: Women recognize circumstances, separate from daily adherence issues, in which they will stop PrEP. These may or may not correspond to lower risk contexts. Counseling is needed when women transition off PrEP to discuss risk and strategies for maintaining sexual health. Proportion Days Covered as a Measure of Adherence in US PrEP Subjects Keith Rawlings, David Magnuson 2, Robertino Mera Giler 2 Gilead Sciences, Medical Affairs, Foster City, CA, United States, 2 Gilead Sciences, Drug Sfty & Public Health, Foster City, CA, United States Background: In 204 the CDC and the WHO recommended the use of once daily Truvada (TVD, tenofovir/emtricitabine) for pre-exposure prophylaxis (PrEP) in combination with other strategies to reduce the risk of sexually acquired HIV- in adults at high risk. A recent analysis of PrEP subjects showed that four doses per week significantly reduced HIV acquisition. The objective here is to evaluate PrEP adherence in a large US administrative database. Methods: The proportion of days covered (PDC) is defined as the percent of days TVD was supplied during the first 80 days of therapy. Only subjects with 2 Rx were included in the analysis. We used national electronic data from ~ 55% of all US retail pharmacies between January 202 and March 204. A previously described algorithm (Mera et al. ICAAC 203) was used to identify TVD for PrEP. Quantile (median) regression was used to estimate the adjusted medians of several risk factors. Results: 8358 prescriptions were analyzed from 3253 unique individuals. A total of 07 individuals met the inclusion criteria. The median PDC was 64.3%, 95% CI %. The multivariate quantile regression showed that gender was significantly associated with median adherence higher in males (77%, 95% CI 73-80%) than females (34%, 95% CI 29-40%; p< 0.0). Adherence also significantly increased by calendar year, with individuals prescribed TVD in 203 having higher medians (70%, 95% CI 65-74%) than those in 202 (50%, 95% CI 46-56%). There was a significant relationship between age and adherence, with older subjects having a higher median PDC (4% more for 0 years difference). Conclusions: A recent analysis suggests that drug levels consistent with 4 TVD pills/wk resulted in a HIV risk reduction of 86-00%. Overall adherence to TVD for PrEP in this retail data base population is consistent with levels that have shown significant reduction in HIV transmission in clinical trials. This is the first assessment of adherence to PrEP among individuals outside of a clinical trial setting. ORAL ABSTRACT SESSIONS 8

82 Oral Abstract Sessions Oral Abstract Session 6: Novel Vaccine Concepts OA6.0 Live Adenovirus Type 4 Recombinants Induce Durable Neutralizing Antibody Responses to Influenza H5 with High Levels of Somatic Hypermutation in Humans Jinghe Huang, Byong Kang, Adam Wheatley 2, Sandeep Narpala 2, Adrian Mcdermott 2, April Poole, Robert Bailer 2, Richard Koup 2, Stephen Migueles, Marc Gurwith 3, Mark Connors NIAID/LIR, HIV-Specific Immunity Section, Bethesda, MD, United States, 2 NIAID, NIH, Vaccine Research Center, Bethesda, MD, United States, 3 PaxVax Inc., La Jolla, CA, United States OA6.02 Non-HIV-derived (Poly)peptides as Primary Immunogens Followed by Envelope Boost Elicit Cross-clade Neutralizing HIV- Antibodies Mei-Yun Zhang, Zheng Yang, Jingjing Li, Qingsheng Liu, Tingting Yuan, Yanyu Zhang, Li-Qing Chen 2, Qi Lou 2, Huazhong Yin 2, Dimiter S. Dimitrov 3 University of Hong Kong, Hong Kong, China, 2 Zhejiang Academy of Medical Sciences, Hangzhou, China, 3 National Cancer Institute, NIH, Frederick, MD, United States ORAL ABSTRACT SESSIONS Background: Induction of durable antibody responses capable of neutralizing diverse HIV- isolates is the most important goal in HIV vaccinology. Broad and potent HIV Env-specific antibodies isolated from humans are characterized by binding to conserved sites on Env and by high levels of hypermutation. Live virus recombinant vectors may provide prolonged exposure to transgene- encoded antigen and may more potently drive affinity maturation than non-replicating vectors. We report results of a small Phase trial of mucosal immunization with live adenovirus type 4 encoding influenza H5 HA (Ad4-H5-Vtn) as a model antigen. We explored its ability to induce durable, broad neutralizing antibody responses through upper respiratory tract application. Methods: Live Ad4-H5Vtn ( viral particles (VP)) was administered by tonsillar swab or by nasal spray in a dose escalation study. Controls received (0 0 VP) as enteric-coated capsules. Neutralizing antibodies were measured by a pseudotype HA lentiviral vector system. B cells were isolated using fluorescent HA probes. Immunoglobulin genes were cloned for mutation analysis, and re-expressed to measure neutralization potency and breadth. Results: The vaccine was well tolerated and symptoms were consistent with mild URI. Tonsillar and nasal immunizations were the most immunogenic routes inducing neutralizing antibody responses at day 60 (ID 50 = 809 and 666 respectively)which rose during the first 6 months. B cells that expanded early were specific for both H and H5 and were directed to the conserved stalk, with H5 HA-monospecific cells expanding by 6 months. The median level of hypermutation was 3.6% compared to germline at week 4 and 7.4% at weeks However, some antibodies contained 3-34% mutation and neutralized group and 2 viruses. Conclusions: A single dose of live recombinant Ad4-H5 can induce highly mutated antibodies, suggesting that mucosal administration is a promising platform for the induction of HIV-specific neutralizing antibodies. Background: A fundamental challenge to develop an effective HIV- vaccine is to identify vaccine immunogens that can initiate and maintain immune responses leading to elicitation of broadly neutralizing HIV- antibodies (bnabs) through complex maturation pathways. We have previously found that HIV- envelope glycoproteins (Env) lack measurable binding to putative germline predecessors of known bnabs and proposed to search for non-hiv-derived immunogens that could initiate their somatic maturation. Methods: Using b2 as a model bnab and yeast display technology, we identified five non-hiv-derived (poly)peptides that bound to its putative germline and intermediate antibody predecessors. Results: Rabbit immunization with the (poly)peptides alone induced high titers of antibodies cross-reactive with Envs, and serum IgGs neutralized SF62 and JRFL. Priming rabbits with the (poly)peptides followed by boosts with an gp40 SF62 trimer and a resurfaced Env (RSC3) induced antibodies that competed with mature b2 for binding to gp40 SF62 trimer and neutralized clade B, C and E isolates. In contrast, rabbits immunized only with gp40 SF62 and RSC3 did not compete with mature b2 for binding to gp40 SF62 trimer and neutralized only clade B isolates. Following the priming with the (poly)peptides, the boosting (secondary) immunogens and the order of different boosting immunogens in rabbit immunization significantly affected the profile and neutralization potency of the induced rabbit antibodies. Conclusions: Our study is the first to provide an evidence that appears to support the concept that non-hiv-derived (poly)peptides may serve as primary immunogens to initiate immune responses leading to elicitation of cross-clade neutralizing antibodies. 82 HIV Research for Prevention 204 HIV R4P

83 Oral Abstract Session 6: Novel Vaccine Concepts Wednesday, 29 October OA6.03 Mechanisms of Protection Observed with Varicella Zoster Virus as a Vaccine Vector in the SIV Macaque Model Catia T. Perciani, David O. Willer 2,3, Kamaran Haq 2,3, Oluwadamilola H. Iwajomo 2,3, Jacqueline K. Chan 2,3, Richard Pilon 4, Joseph M. Antony 2,3, Paul Sandstrom 4, Kelly S. Macdonald,2,3 University of Toronto, Department of Immunology, Toronto, ON, Canada, 2 University of Toronto, Department of Medicine, Toronto, ON, Canada, 3 Mount Sinai Hospital, Department of Microbiology, Toronto, ON, Canada, 4 Public Health Agency of Canada, National HIV and Retrovirology Laboratories, Ottawa, ON, Canada OA6.04 Genetic Assessment of B-cell Responses Following Immunization with HIV Env Protein and Adjuvants or SHIV (AD8) Infection in Nonhuman Primates Joseph Francica, Zizhang Sheng 2, Yoshiaki Nishimura, Zhenghai Zhang 2, Manmohan Singh 3, Derek T. O Hagan 3, Daniel Douek, Thomas Kepler 4, Malcolm Martin, Lawrence Shapiro 2, Robert Seder NIH, Bethesda, MD, United States, 2 Columbia University, New York, NY, United States, 3 Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United States, 4 Boston University, Boston, MA, United States Background: Among the HIV-vaccine strategies tested to date, those involving recombinant viral vectors with ongoing replication such as those from the Herpesvirus family show the most promise. We have taken advantage of a well studied virus, Varicella Zoster Virus (VZV), as a vector for HIV antigens and anticipate the induction of a durable and multifaceted immune response against HIV when adopting this approach. VZV establishes a latent infection with cycles of sub-clinical reactivation that can potentially boost the immune response against HIV antigens throughout life; an advantage over prime-boosting schemes against HIV. Methods: We have recently utilized VZV as an SIV vaccine vector in the cynomolgus macaque model of HIV and demonstrated that approximately 33% of the animals vaccinated a single time with a VZV-SIV construct and challenged intra-rectally with multi low-dose SIV, controlled SIV viral load down to undetectable levels after initial infection. In addition, the remaining vaccinees maintained their setpoint viral loads significantly lower than controls (p< 0.00). Immune responses associated with viral control have been assessed by multiparameter flow cytometry in various tissues including blood, rectal tissue, lymph nodes and broncho-alveolar sites. Results: Analysis of the systemic immune response showed the presence of a pool of T EM cells that correlated to a reduction in the set-point viral load (p< 0.00). We also have found evidence that Th7 response in the gut and lymph nodes may play an important role in the control of viral load (p< 0.05). We are currently investigating the dynamics between different T cell populations (T EM, Tregs, Th7 and Th22) as well as immune activation longitudinally during the infection phase of this trial with emphasis on the interplay between blood and mucosa. Conclusions: The sustained viral clearance observed in this study offers an exciting platform to investigate possible immune correlates of protection against SIV. Background: An effective preventive vaccine for HIV- should generate potent neutralizing antibody responses against the HIV envelope (Env). However, broadly-neutralizing antibodies from HIV+ individuals often have unique characteristics such as extensive somatic hypermutation (SHM), long CDRH3 regions and V H gene restriction. Therefore, we monitored the development of these characteristics in the preclinical setting. Methods: NHP were immunized with HIV Env clade C protein alone, or with a panel of clinical adjuvants or were infected with SHIV(AD8) virus. Env-specific B cells were sorted, barcoded, deep sequenced, and analyzed using a newly curated NHP Ig reference database. In total we analyzed V H gene usage, SHM, and CDRH3 length from over 58,000 unique Ig heavy chains. Results: The adjuvants tested here did not significantly increase germline divergence compared to unadjuvanted protein, nor did they give rise to B cells with long CDRH3 regions. However, animals and individual reads with higher SHM showed evidence of a narrowed V H gene repertoire and were enriched in the IGHV3Q gene. By comparison, antibodies from SHIV infected animals showed an increase in SHM in animals with better neutralization responses. Better neutralization responses further correlated with higher viral loads and viral Env diversity, indicating that chronic antigen stimulation and diversity is likely a driving force for higher SHM and the development of antibody potency and breadth. Finally, we correlated the presence of 3 V H genes: IGHV4A, 4D and 3J with better neutralization, indicating it may be possible to identify a V H gene signature of neutralization. Conclusions: This study is the first comprehensive comparison of antibody genetics using vaccine and infection models in NHP. Our data suggest that future vaccine regimens should be designed to mimic chronic viral infection, utilizing heterologous Env immunogens and prolonged innate and antigen stimulation to drive affinity maturation and neutralization breadth. ORAL ABSTRACT SESSIONS 83

84 Oral Abstract Sessions Oral Abstract Session 6: Novel Vaccine Concepts OA6.05 Expanded Epitope Recognition by Vaccination with DNA Encoding Novel Immunogens Comprising HIV Conserved Elements OA6.06 Vaccine Induced Distinct Circulating Memory CD4+ T-cells with T-follicular Helper Cell Commitment in Humans ORAL ABSTRACT SESSIONS Barbara K. Felber, Xintao Hu, Viraj Kulkarni, Antonio Valentin, Margherita Rosati, Candido Alicea, Morgane Rolland 2, Sylvie Le Gall 3, Niranjan Y. Sardesai 4, Beatriz Mothe 5, Christian Brander 5,6, James I. Mullins 2, George N. Pavlakis National Cancer Institute at Frederick, Frederick, MD, United States, 2 University of Washington, Seattle, MD, United States, 3 Ragon Institute of MGH, MIT, and Harvard, Boston, MD, United States, 4 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States, 5 IrsiCaixa AIDS Research Institute - HIVACAT, Autonomous University of Barcelona, Barcelona, Spain, 6 Institucio Catalana de Recerca i Estudis Avancats (ICREA), Barcelona, Spain Background: HIV sequence diversity and potential decoy epitopes are hurdles in the development of an effective AIDS vaccine. To target immune responses towards invariable viral regions we engineered DNA-based immunogens encoding conserved elements (CE) of HIV- selected on the basis of stringent conservation, functional importance, and association with immune control. Methods: Macaques were immunized by IM injection followed by electroporation with DNA plasmids expressing CE or complete immunogens (gag, env), alone or in heterologous prime-boost regimens. Immune responses elicited by CE from gag (HIV, SIV) and env (HIV) were compared to those obtained upon vaccination with DNA expressing the complete antigens. Results: Macaques vaccinated with CE gag or CE env DNA developed robust CE-specific cytotoxic T cells (Granzyme B+, CD07+). CE-specific T cells were found only in ~50% of animals vaccinated with DNA encoding the complete antigen and these responses targeted fewer CE per animal. Importantly, boosting CE-primed macaques with DNA expressing full-length antigen increased both magnitude and breadth of the CE responses. Although designed as CTL vaccine, p24ce DNA vaccine induced Ab responses able to recognize p55 gag and targeting a broad range of linear epitopes. In contrast, antibodies induced by p55 gag DNA vaccination failed to recognize p24ce or linear CE epitopes. Interestingly, boosting of p24ce DNA primed macaques with p55 gag DNA increased Abs recognizing HIV p24 gag as well as p24ce proteins, thereby inducing broadest immunity. Conclusions: Combination of conserved elements and full-length immunogen provides a novel strategy to increase the magnitude and breadth of cellular and humoral immunity while targeting efficiently the conserved regions of the virus. This allows for the development and expansion of subdominant responses and greater breadth of immune response. Antje Heit, Frank Schmitz 2, Miranda Moore 3, Sarah Gerdts 3, Britta Flach 3, Harlan Robins 3,4, Alan Aderem 2,5, Stephen de Rosa 3,5, M Juliana McElrath 3,5 Fred Hutchinson Cancer Research Center, VIDD, Seattle, WA, United States, 2 Seattle Biomedical Research Institute, Seattle, WA, United States, 3 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 Adaptive Biotechnologies, Seattle, WA, United States, 5 University of Washington, Seattle, WA, United States Background: Peripheral blood CD4+ T follicular helper (ptfh) cells were recently identified in humans, but it is unclear whether they resemble a stable population of antigen-specific memory cells responsive upon antigen re-encounter. We investigated ptfh kinetics, population diversity and their relationship to B-cell responses in vaccine recipients. Methods: PBMC samples, longitudinally collected from volunteers enrolled in HVTN068 from either the DNA/Ad5 or Ad5/Ad5 arm as well as from healthy, non-vaccinated donors were used to identify three distinct CXCR5+ ptfh subsets by their individual expression pattern of PD and ICOS. Multicolor flow cytometry, qpcr and deep sequencing of the TCRB repertoire of individual CD4+ T cell subsets was applied to characterize and phenotype the individual ptfh subsets. Results: Amongst the different ptfh subsets, the vaccine-specific PD/ ICOS double-positive memory ptfh (ptfh dp ) showed a unique clonal expansion one week after boosting. Simultaneously, vaccine-specific, IgG expressing plasmablasts appeared in the circulation. At the same time, proliferating ptfh dp cells up-regulated germinal center Tfh genes, and became enriched for Th-like cells. Strikingly, during the peak response the T-cell receptor clonal repertoire of ptfh dp was completely exchanged and identical T-cell clones were shared with PD+ ICOSmemory ptfh (ptfh PD ) cells. Conclusions: We conclude that vaccination induces memory ptfh PD that respond with a rapid but transient expansion and differentiation into activated ptfh dp following antigen reencounter. 84 HIV Research for Prevention 204 HIV R4P

85 Oral Abstract Session 7: Mucosal Target and Effector Cells Wednesday, 29 October OA7.0 Immune Activation and HIV Target Cells in the Adolescent Female Genital Tract OA7.02 Functional Dynamics of the Interaction between HIV gp20 and α4β7 Smritee Dabee, Heather B. Jaspan,2, Shaun L. Barnabas,3, Shameem Z. Jaumdally, Hoyam Gamieldien, David Lewis 4,5, Thola Bennie 3, Angel Phuti 3, Clive M. Gray,6, Anna-Lise Williamson,6, Thomas J. Hope 7, Francesca Chiodi 8, Robin Shattock 9, Jo-Ann S. Passmore,6, Linda-Gail Bekker 3,0, Women s Initiative in Sexual Health (WISH) Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Department of Clinical Laboratory Sciences, Cape Town, South Africa, 2 Seattle Biomed, Seattle, WA, United States, 3 Desmond Tutu HIV Foundation, Cape Town, South Africa, 4 Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Department of Microbiology, Cape Town, South Africa, 5 National Institute for Communicable Diseases, Johannesburg, South Africa, 6 National Health Laboratory Services, Cape Town, South Africa, 7 Feinberg School of Medicine, Northwestern University, Department of Cell and Molecular Biology, Chicago, IL, United States, 8 Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 9 Imperial College London, Department of Infectious Diseases, London, United Kingdom, 0 Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa Background: The biological mechanisms underlying HIV risk in younger women is unclear. As HIV is primarily transmitted across the genital mucosa and preferentially infects activated CD4 + cells, we investigated the influence of age and sexually transmitted infections (STIs) on CD4 + target cell immune activation from the genital tract of adolescent females from South Africa. Methods: As part of a longitudinal cohort study aiming to enrol 50 adolescent women between the ages of 6-22 years at the Desmond Tutu Youth Centre, Masiphumele, Cape Town [part of the EDCTP funded Women s Initiative in Sexual Health (WISH)], cervical mononuclear cells were obtained from 35 adolescents and the T-cell expression of activation and proliferation markers (CD38, HLA-DR, Ki67) was measured by FACS. Women were screened for bacterial vaginosis (BV) and STIs (C. trachomatis, N. gonnorhoea, T. vaginalis, M. genitalium, HSV- & 2 and HPV). For comparison, HIV-negative adult women were also included. Results: Adolescents (median 8 years; IQR 8-9) had significantly higher frequencies of activated CD4 + T-cells (CD38 +, HLADR +, CD38 + HLADR + : each p< 0.000) than adult women. In contrast, adolescents had significantly lower frequencies of CD4 + T-cells expressing CCR5 (p=0.02) than adult women. When adolescents were stratified according to age (6-7, 8-9 and year old), the 6-7 year old females generally had higher levels of activated and proliferating T-cells compared to the other age groups, significantly so for the proliferating CD4 + T-cells (Ki67+; ANOVA: p=0.048). The prevalence of STIs and BV were high in adolescents, with 40% testing positive for C. trachomatis. Adolescents infected with C. trachomatis tended to have higher levels of T-cell activation compared to those without an STI, significantly so for CD38 + HLADR + T-cells (p=0.0). Conclusions: Heightened levels of genital immune activation found in South African adolescent females, partly due to the presence of STIs, could put them at higher risk of HIV infection. Fatima Nawaz, Claudia Cicala, Katija Jelicic, Raffaello Cimbro 2, Donald Van Ryk, Jocelyn Ray, Joseph Hiatt, Catherine Schwing, Danlan Wei, Massimiliano Pascuccio, Aftab A. Ansari 3, Anthony S. Fauci, James Arthos NIAID, National Institutes of Health, Bethesda, MD, United States, 2 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 3 Emory University School of Medicine, Atlanta, MD, United States Background: Many isolates of HIV and SIV exhibit a specific affinity for integrin α4β7, the gut homing receptor. CD4+ T cells expressing α4β7 represent an ideal target for productive infection during mucosal transmission. However, many details surrounding the interaction between HIV and α4β7 remain unresolved. We hypothesize that the evolution of a specific interaction between HIV and α4β7 provides the virus a means to increase the efficiency of infection in mucosal tissues. Disrupting this interaction may provide a strategy to reduce or prevent mucosal transmission. Here we identify key features of the α4β7-hiv gp20 interaction that facilitate viral replication. Methods: We evaluated the capacity of gp20, by binding to α4β7, to induce cellular proliferation of primary CD4+ T cells using CFSE proliferation assays. We also examined the manner in which α4β7 is physically linked to CD4 using high-resolution confocal microscopy and fluorescence resonance energy transfer (FRET) on primary CD4+ T cells. Results: We find that CD4 and α4β7 are closely associated (< 0nM) on the surface of CD4+ T cells. Both gp20 and soluble MAdCAM mediated costimulation of cellular proliferation in an α4β7-dependent manner. Conclusions: Although α4β7 is not an entry receptor we find that it is closely associated with CD4 on the cell membrane. This association suggests that α4β7 plays a role in T cell proliferation. We determined that both gp20 and MAdCAM costimulate CD4+ T cells and facilitate cellular proliferation and viral replication. We hypothesize that one of the selective pressures leading to a specific interaction between gp20 and α4β7 involves the capacity of α4β7 to promote metabolic activity that is conducive to viral replication. In the context of mucosal transmission, that is generally inefficient, these features of HIV interactions with α4β7- provide a means to increase transmission efficiency. Future studies will address the potential of targeting this interaction as a means of interfering with transmission. ORAL ABSTRACT SESSIONS 85

86 Oral Abstract Sessions Oral Abstract Session 7: Mucosal Target and Effector Cells OA7.03 OA7.04 ORAL ABSTRACT SESSIONS Preferential HIV Susceptibility of Specific CD4+ T Cell Subsets in the Cervix Vineet R. Joag, Lyle R. McKinnon 2,3, Segen Kidane, Mark Yudin 4, Steve Kimwaki 5, Stephanie Rainwater 6, Julie Overbaugh 6, Omu Anzala 5,7, James Arthos 8, Joshua Kimani 5, Rupert Kaul,2,5 University of Toronto, Immunology, Toronto, ON, Canada, 2 University of Toronto, Medicine, Toronto, ON, Canada, 3 CAPRISA, Durban, South Africa, 4 University of Toronto, Obstetrics and Gynecology, Toronto, ON, Canada, 5 University of Nairobi, Medical Microbiology, Nairobi, Kenya, 6 University of Washington, Fred Hutchinson Cancer Research Centre, Seattle, WA, United States, 7 University of Nairobi, Kenya AIDS Vaccine Initiative, Nairobi, Kenya, 8 National Institute of Allergy and Infectious Diseases, Bethesda, MD, United States Background: A better understanding of the cellular targets of HIV in the female genital tract may inform the development of HIV vaccines and microbicides. Proposed correlates of cellular susceptibility include expression of the HIV co-receptor CCR5, mucosal homing integrins, and/ or CD69, a marker of early immune activation. Methods: We used a CCR5-tropic HIV pseudovirus to quantify HIV entry into unstimulated cervical CD4 T cell subsets in a rapid, flow cytometrybased assay. We also assessed the metabolic state of cervical cell subsets based on Ki67 expression, since proliferating cells are more susceptible to productive HIV infection. Results: HIV entry into cervical T cells was CD4 and CCR5 dependent. Viral entry was approximately three-fold higher into cervical CD4 T cells compared to blood (p=0.0003), although within a given participant, virus entry was strongly correlated between the two compartments (r=0.748, p=0.0002), suggesting shared host determinants of HIV entry. Cervical CD4 T cells that expressed a4b7 (p=0.003), a4b (p=0.0002), or CD69 (p=0.0004) were approximately two-fold more susceptible to HIV entry; CCR5 expression was increased at least two-fold on a4b7+ (p< 0.000) and CD69+ (p< 0.000) CD4 T cells, but only.2 fold on a4b+ (p=0.0002) CD4 T cells. Cervical a4b7+ CD4 T cells expressed two-fold higher levels of the proliferation marker Ki67 (p=0.003), which was not seen in CCR5+ (p=0.95), CD69+ (p=0.46) or a4b+ (p=0.24) CD4 T cells. Conclusions: Preferential HIV entry into immunologically activated and a4b7+ cervical CD4 T cells was probably due, at least in part, to increased CCR5 expression. Increased HIV entry into cervical a4b+ CD4 T cells did not correlate with CCR5 overexpression, and the mechanism of enhanced cell entry in this cell subset is unknown. Markers of metabolic activity and proliferation were specifically increased in cervical a4b7+ CD4 T cells suggesting that this subset may be particularly suited to sustain productive HIV infection. Expression of MAIT Cells in Blood and Genital Mucosa of HIV Infected and Uninfected Women Anna Gibbs, Edwin Leeansyah 2, Andrea Introini, Taha Hirbod, Joshua Kimani 3, Terry B. Ball 4,5, Kristina Broliden, Johan K. Sandberg 2, Annelie Tjernlund Karolinska Institutet, Department of Medicine, Solna, Stockholm, Sweden, 2 Karolinska Institutet, Department of Medicine, Huddinge, Stockholm, Sweden, 3 University of Nairobi, Department of Medical Microbiology, Nairobi, Kenya, 4 University of Manitoba, Department of Medical Microbiology, Winnipeg, MB, Canada, 5 Public Health Agency of Canada, National Microbiology Laboratory, Winnipeg, MB, Canada Background: Mucosa-associated invariant T (MAIT) cells are a recently described antimicrobial T cell population, which is abundant in blood and mucosal surfaces. MAIT cells have been shown to be reduced in blood, however preserved in mucosa of HIV infected patients. The aim of this study is to define number, localization, and functional profile of MAIT cells ex vivo in tissue samples from the female genital tract (FGT) of HIV infected and uninfected women. Methods: Cervical tissue samples were collected from Swedish women undergoing hysterectomy (n=5), and paired PMBC and ectocervical biopsies were collected from HIV seropositive female sex workers (n=20), HIV seronegative female sex workers (n=7), and HIV seronegative lower risk women (n= 2) from Kenya. An advanced multicolor flow cytometry panel was used to enumerate MAIT cells and determine their functional profile. In situ staining was performed to define the distribution of the MAIT cells in the tissue samples. Results: MAIT cells represent 2% out of the total amount of mucosal T cells in the cervical and endometrial compartments of the Swedish HIV uninfected women. The majority of the MAIT cells are CD8 + T cells and they produced high levels of IFN- γ, TNF, IL-7, Granzyme B, and moderate levels of IL-22 after E. coli stimulation. MAIT cells were located close to the basal membrane of ectocervix and in the submucosal compartment of the endometrium. We will evaluate the samples from the Kenyan women in regards to the percentage of MAIT cells, in blood and in the mucosa, of HIV infected vs. uninfected women. Conclusions: MAIT cells are present in the FGT of Swedish HIV uninfected women and they display microbial reactivity by producing pro-inflammatory cytokines and cytolytic molecules. The production of IL-7 and IL-22 may allow MAIT cells to play a role in regulating homeostasis and integrity of the mucosal barrier in FGT. This may be an important mechanism to defend the mucosal genital barrier integrity and prevent dissemination of local infections. 86 HIV Research for Prevention 204 HIV R4P

87 Oral Abstract Session 7: Mucosal Target and Effector Cells Wednesday, 29 October OA7.05 OA7.06 Rapid Loss of Th7 Cells after SIV Infection May Underlie Mucosal Dysfunction Laura E. Richert-Spuhler, Tiffany Hensley-Mcbain, Jillian Gile, Michael Koday, Deborah H. Fuller, Brian Johnson 2, Melon T. Nega 3, Cara Appel 2, Thomas Vanderford 3, R. Keith Reeves 4, Jacob D. Estes 5, Brandon F. Keele 5, Nichole R. Klatt University of Washington, Pharmaceutics and Washington National Primate Research Center, Seattle, WA, United States, 2 University of Washington, Department of Comparative Medicine, HIC/Comparative Pathology Program, Seattle, WA, United States, 3 Emory University, Emory Vaccine Center, Yerkes National Primate Research Center, Atlanta, GA, United States, 4 Harvard University, Beth Israel Deaconess Medical Center, Boston, MA, United States, 5 Frederick National Laboratory for Cancer Research, AIDS and Cancer Virus Program, SAIC- Frederick, Inc., Frederick, MD, United States Background: During chronic HIV/SIV infection, CD4+IL-7-producing T cells (T H 7) are significantly depleted from mucosal tissues, and their absence is highly associated with GI dysfunction. However, the kinetics of immune dysfunction, in sequence with microbial translocation and inflammation remain unknown. Herein we elucidate the kinetics of acute host/pathogen interactions with the goal of identifying early events after HIV infection that reveal potential prevention strategies. Methods: We collected longitudinal biopsies and blood at early acute timepoints from six rhesus macaques following intrarectal SIV challenge with SIVmac239x (00,000 TCID 50 ). Rectum, colon, jejunum, lymph nodes, and blood were collected pre-siv (day -2) and days 3, 7, 4, 28, 42, and 63 post-challenge. Immunophenotype, functionality, inflammation, virus kinetics, and microbial translocation were measured. Results: Strikingly, T H 7 cells were significantly depleted from all GI tract sites by day 3 post-siv infection (rectum: p=.005; colon: p=.032; jejunum: p=.047). This depletion was neither correlated to generalized CD4 depletion, nor to generalized loss of cytokine production (exhaustion/anergy). The marked loss of T H 7 cells appeared to be unique to this particular subset, as we did not observe dramatic early loss of IL-7+CD8+ T cells or innate lymphoid cells. Additionally, we found that loss of T H 7 cells preceded overt inflammation and microbial translocation, indicating that this deficiency may underlie subsequent mucosal dysfunctions observed during HIV/SIV. Conclusions: Taken together, our data indicate that the loss of T H 7 cells from the GI tract at the earliest stages of infection uniquely precedes microbial translocation and a systemic proinflammatory state. The preservation of T H 7 cells may, therefore, be an important factor in preventing HIV transmission and pathogenesis. Furthermore, targeted T H 7 cell retention could serve as a novel inhibitory strategy against HIV pathologies. No Increase in Activated T Cells and Limited Increase in Adenovirus-specific T Cells in Colon and Rectal Mucosa Following HIV- DNA/rAd5 Immunization Stephen De Rosa, Nicole Frahm, Gabriela Diaz, Paul Newling, Daryl Morris, Cecilia Morgan, Barney Graham 2, Edith Swann 3, Georgia Tomaras 4, Janine Maenza, Ann Duerr, M Juliana McElrath Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 Vaccine Research Center, NIH, Bethesda, MD, United States, 3 DAIDS/ NIAID/NIH, Bethesda, MD, United States, 4 Duke University, Durham, NC, United States Background: HVTN076 is an exploratory vaccine trial examining mucosal immune responses to the NIH Vaccine Research Center multiclade HIV- DNA/rAd5 vaccine, also used in the HVTN505 test-ofconcept study. Methods: 7 healthy Ad5 seronegative HIV-uninfected adults enrolled; 6 completed all vaccinations (3xDNA, rad5; 0, 4, 8, 24 wks). Mucosal biopsies were obtained from sigmoid colon and rectum by flexible sigmoidoscopy at baseline and month after rad5. Rectal biopsies by anoscopy were obtained 2 weeks after final DNA and week after rad5. Single-cell suspensions following collagenase digestion were assayed directly for phenotyping and after overnight rest for intracellular cytokine staining. HIV-specific IgG was measured in serum and mucosal secretions. Results: HIV-specific CD4+ and CD8+ T cells were detected in blood at month post rad5 (69 and 44% of participants for Gag, respectively), but none were detected in colon/rectum samples at any timepoint. Ad-specific CD4+ T cells, measured in response to the hexon coat protein, were detected at baseline in blood (92%) and colon (87%). Hexon-specific response rate and magnitude increased in blood after vaccination, but this increase was less common in colon/rectum. The proportion of CD4+ T cells expressing CCR5 or activated CD4+ T cells (Ki-67+BcL-2lo) in the rectum was unchanged after rad5 vaccination. Env-specific IgG was detected in serum and mucosal secretions. Conclusions: The vaccine regimen did not induce detectable HIV-specific T cells in colonic/rectal mucosa, an important site for HIV acquisition. These cells may be present, but require in vitro expansion for detection. Pre-existing mucosal Ad-specific T cells were detected in the majority of these Ad5 seronegatives. Activation of such cells is a potential concern for enhanced risk of HIV infection after rad5 vaccination; however, postvaccination increase in Ad-specific T cells was detected in few vaccinees. Vaccine-induced activation of T cells in bulk, as measured by Ki-67/ BcL-2, was not detected. ORAL ABSTRACT SESSIONS 87

88 Oral Abstract Sessions Oral Abstract Session 8: Evaluation of Novel Interventions OA8.0 OA8.02 ORAL ABSTRACT SESSIONS HbAHP-25 Inhibits HIV- Entry into Cells and Neutralizes gp20 Induced Inflammation Tahir Bashir, Mandar Patgaonkar 2, Selva Kumar 3, KVR Reddy National Institute for Research in Reproductive Health [NIRRH], Division of Molecular Immunology & Microbiology, Mumbai, India, 2 Tata Institute of Fundamental Research, Department of Biological Sciences, Mumbai, India, 3 Dr. D.Y. Patil University CBD Belapur, Department of Bioinformatics, Mumbai, India Background: HIV/AIDS is one of the major causes of death worldwide. Most of the new HIV infections occur through heterosexual mode of transmission. Hence, preventing HIV infection at this portal of entry is pivotal in eradicating AIDS epidemic. Methods: HbAHP-25, a peptide against CD4 binding domain of gp20, was designed in silico by molecular docking using Z-dock software. Binding of this peptide to gp20 was evaluated using ELISA and flow cytometry. Activity of HbAHP-25 against various strains of HIV- was evaluated on TZM-bl cells and PBMCs using Luciferase and p24 assay in presence of seminal plasma and vaginal fluid. Competitive ELISA was performed to determine effect of HbAHP-25 on CD4 binding to gp20. Cell viability and epithelial monolayer integrity in presence of HbAHP-25 was determined by MTT assay and Immunofluorescence. Multiplex cytokine assay was performed to determine inflammatory status of peptide and its role in gp20 mediated inflammation. Results: HbAHP-25 exhibits significant anti-hiv activity against various strains of HIV- in a dose dependent fashion. Results from ELISA and SPR reveal direct interaction between HbAHP-25 and gp20; it binds to a site proximal to CD4 binding site on gp20, and has partial epitope similarity with VRC0. HbAHP-25 retains its anti-hiv potential in presence of vaginal fluid & seminal plasma. It didn t affect cell viability/ monolayer integrity even at concentrations 4-5 fold higher than its IC 50. HbAHP-25 didn t affect tight junction proteins and did not show any significant induction of pro/anti-inflammatory cytokines; instead it reduced gp20 induced activation of pro-inflammatory cytokines viz., IL-α, IL-6, IL-8 and GM-CSF. Conclusions: HbAHP-25 has potent anti-hiv activity by blocking gp20- CD4 interaction and alleviating gp20 induced inflammation in vaginal cells, suggesting its plausibility as a potential prophylactic/microbicidal agent. Binding site of HbAHP-25 on gp20 can also be a potential site for vaccine development. Bypass of Quality Control in Protein Folding Pathways Induces Specific Misfolding of HIV Envelope V2 Loop: Implications for Iminosugars as Antivirals Simon Spiro, Nicholas McCaul 2, Ronald Derking 3, Dominic Alonzi, Raymond Dwek, Rogier Sanders 3, Ineke Braakman 2, Nicole Zitzmann University of Oxford, Institute of Glycobiology, Oxford, United Kingdom, 2 University of Utrecht, Cellular Protein Chemistry, Utrecht, Netherlands, 3 University of Amsterdam, Department of Medical Microbiology, Amsterdam, Netherlands Background: The HIV Env glycoprotein is vital for cell targeting and entry. During folding in the endoplasmic reticulum the sequential trimming of glucose residues from the protein s glycans by ER α-glucosidases is crucial in targeting it to the chaperones calnexin and calreticulin (CNX/CRT). Iminosugars are a class of glucose-mimetic drugs that inhibit α-glucosidases preventing association of Envelope with CNX/ CRT. NB-DNJ is an iminosugar that is antiviral against all clades of HIV and no escape mutants have ever been detected even after months of treatment in vitro. We used NB-DNJ to study the role of CNX/CRT in Env folding and to better understand its antiviral functions. Methods: We used radioactive pulse-chase labelling to study the effects of NB-DNJ on Env oxidative folding in the ER and then conformational monoclonal antibody binding to examine the structure of the final Env protein, using both gp20 and BG505 SOSIP trimers. We also used molecular clones to study the effects of the drugs on the production of whole viral particles and derivative mutants. Results: In the presence of NB-DNJ nascent gp20 oxidised unusually rapidly, after which there was a slow isomerisation of the disulphides back into an apparently native conformation. However, antibody mapping of the secreted protein showed the V2 loop was in an aberrant conformation. HIV mutants that lacked the V/V2 loop were partially resistant to the effects of the drug. While the proportion of misfolded protein correlated well to α-glucosidase inhibition, the antiviral effect did not- instead full suppression of virion infectivity occurred when only a small proportion of the proteins were misfolded. Conclusions: This lack of correlation can be explained by the need for HIV envelope proteins to trimerise and then cluster in order to function; the misfolding of a single protomer could then affect the greater whole, thus explaining the selectively antiviral effects of iminosugars. CNX/CRT is vital for the folding of V/V2 but not for the rest of Env. 88 HIV Research for Prevention 204 HIV R4P

89 Oral Abstract Session 8: Evaluation of Novel Interventions Wednesday, 29 October OA8.03 OA8.04 A Role for Scavenger-like Lymphocyte Receptor CD6 in HIV- Viral Infection Esther Carrasco, Cristina Escoda, Carmen Alvarez-Fernández 2, Sonsoles Sanchez-Palomino 2, Esther Carreras, José Maria Gatell 2,3, Teresa Gallart 4, Felipe García 2,3, Núria Climent 2, Francisco Lozano,4,5 Immunoreceptors Group, Institut d Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Centre Esther Koplovitz, Barcelona, Spain, 2 CELLEX-IDIBAPS-HIVACAT, Barcelona, Spain, 3 Infectious Diseases and AIDS Unit, Hospital Clínic de Barcelona, Barcelona, Spain, 4 Hospital Clínic de Barcelona, Department of Immunology, Barcelona, Spain, 5 Faculty of Medicine, Universitat de Barcelona, Department of Cellular Biology, Immunology and Neuroscience,, Barcelona, Spain Background: CD6 is a lymphocyte receptor that physically colocalizes with the TCR/CD3 complex at the immunological synapse where interacts with CD66/ALCAM, an adhesion molecule of the Ig superfamily. The extracellular region of CD6 exihibits three SRCR (scavenger receptor cysteine-rich) domains, a highly conserved module of the innate immune system. Accordingly, CD6 allow lymphocytes to sense the presence of bacterial cell wall components, and soluble CD6 (scd6) improves mouse survival following lethal bacterial-challenge. Our objective was to analyze whether CD6 would recognize HIV- structures. Methods: scd6 neutralization assays were performed in PBMC using recombinant NL4.3-Renilla (X4) virus. Recombinant gp20, scd6 and scd4 binding was analyzed by ELISA and Western blot. Overlapping 5-mer peptides derived from HIV- MN strain gp20 were obtained from the NIH and used for ELISA competition assays. CD6 and CD4 receptors colocalization assays were analyzed by confocal microscopy. The scd6 mediated inhibition of gp20 binding to human PBMC was analyzed by FACS. Results: We report that scd6 significantly inhibits HIV- infectivity (90% at a dose of 0µg/ml) (p< 0.0). Additionally, scd6 binds to HIV- gp20 in a dose-dependent manner. The CD6-binding region is localized to a linear sequence of the V3 loop important for chemokine receptor interaction (P6283). The interaction of scd6 with gp20 is enhanced by prebinding of scd4 to gp20 (p< 0.00) suggesting that scd6 inhibitory activity is mediated by blocking the gp20/coreceptor interaction. In addition, we demonstrate the interaction and colocalization between CD6 and the CD4 receptor. Finally, scd6 inhibited the gp20 binding to human PBMC, in a dose-dependent manner (p< 0.05). Conclusions: Ours results suggest that scd6 directly interacts with V3 loop of HIV- gp20 and CD4 receptor and likely mediates anti-hiv- activity playing a role in the virological synapse. Furthermore, scd6 could prevent HIV entry being useful as a new HIV- treatment. Testing a Fast-dissolving Tablet Containing a Recombinant Live Biotherapeutic Product, MucoCept-CVN, in the Non-human Primate Model for Colonization Laurel Lagenaur,2, Peter Lee 2, Thomas Parks 2 NCI, Bethesda, MD, United States, 2 Osel Inc., Mountain View, CA, United States Background: Osel developed a recombinant live biotherapeutic product, MucoCept-CVN based on human vaginal Lactobacillus jensenii expressing the HIV- entry inhibitor Cyanovirin-N for prevention of vaginal transmission of HIV-. We previously reported that macaques colonized with a live bacterial preparation in hydroxyethylcellulose showed a 63 % reduction of SHIV SF62P3 acquisition followed repeated vaginal challenge. We formulated MucoCept-CVN as a fast dissolving vaginal tablet, which would allow for discreet, coital-independent, female controlled use. Tablets were potent (5.8 x 0 colony forming units (CFUs)/gram) and stable for at least one year at 4 o C and 25 o C. Macaques were dosed with tablet or 5 tablets delivered over 5 days. We examined colonization of the MucoCept-CVN bacteria in the rhesus macaque vagina at 4 and 2 days post dosing and found that 83% were highly colonized. Methods: Fast dissolving tablet were formulated (~0 mg in a x.5 cm mold). Tablet friability and dissolution were tested. Stability of the product was monitored for year at 4 o C and 25 o C and for 2 months at 37 o C. A single tablet or multiple tablets were administered to 8 rhesus macaques and vaginal colonization was monitored at 4 and 2 days post dosing by culture and PCR. Results: Tablets were potent and contained 5.8 x 0 colony forming units (CFUs)/gram. The tablets were stable yr at 4 o C and 25 o C and for month at 37 o C. The tablets maintained integrity when handled and dissolved quickly. High levels of colonization ~0 8 CFU/vaginal swab of colonization were achieved in 5/8 macaques by day 2 post dosing. Two macaques had lower colonization levels ~0 5 CFU/swab and in one macaque bacteria were detected by PCR only. Conclusions: MucoCept-CVN can be formulated and delivered as a conventional fast dissolving tablet dosage form. Colonization was achieved in 83% of macaques. The tablets were easy to handle and stable for > yr. This tablet formulation can be manufactured on large scale for ease of use. ORAL ABSTRACT SESSIONS 89

90 Oral Abstract Sessions Oral Abstract Session 8: Evaluation of Novel Interventions OA8.05 The Role of Semen on Vaginal HIV- Transmission and on the Efficacy of Maraviroc as a Topically Applied Microbicide Olivia Snyder, Angela Wahl, Michael Swanson, Rae Ann Spagnuolo, J. Victor Garcia UNC Chapel Hill, Chapel Hill, NC, United States OA8.06 Post Coital Assessment of Topical Microbicide Formulations in the Macaque Model Dorothy L. Patton, Yvonne Sweeney, Lisa C. Rohan 2,3 University of Washington, Obstetrics & Gynecology, Seattle, WA, United States, 2 University of Pittsburgh, School of Pharmacy, Pittsburgh, PA, United States, 3 Magee Women s Research Institute, Pittsburgh, PA, United States ORAL ABSTRACT SESSIONS Background: All mucosal exposures to HIV occur in the presence of semen. Currently, there is no consensus on the effect of semen on HIV transmission or on the potential effectiveness of topical microbicides. Here, we use an in vivo animal model of mucosal HIV transmission to establish the effect of semen on vaginal HIV infection and on the efficacy of topical microbicides. Methods: We utilized bone marrow/liver/thymus (BLT) humanized mice; a model validated for the study of vaginal HIV transmission and HIV prevention strategies. We first evaluated the transmission of transmitted/ founder viruses in the presence or absence of human semen. In addition, we also evaluated the effect of semen on cell-associated HIV transmission. Lastly, we evaluated the efficacy of topically applied microbicides in the presence of semen using the CCR5 antagonist, maraviroc. Log rank Mantel-Cox was used to analyze the data. Results: To determine the effect of semen on vaginal transmission HIV- CH040, a transmitted founder (T/F) virus, was resuspended in human semen and vaginally administered to BLT mice. Efficient transmission was observed regardless of the presence (6/6) or absence (4/4) of semen. No differences were noted in the levels of peripheral viral load or CD4+ T cell decline between the two groups. When cell-associated HIV was used for challenge in the presence of semen, 3/4 BLT mice became infected compared to 4/4 in the control arm. When animals were treated vaginally with maraviroc and then challenged with HIV in semen, complete protection was observed (6/6). Conclusions: Our results demonstrate that semen does not enhance transmission of either cell-free or cell-associated HIV. In addition, semen does not diminish the protective effect of maravioc from vaginal HIV infection when applied topically. Our results establish a new paradigm for the evaluation of HIV prevention strategies that includes human semen in the context of cell-free and cell-associated virus. Background: The pigtailed macaque post-coital model provides assessment of the safety of developing topical microbicide products with daily use and with coitus. We have conducted studies using the postcoital safety model to establish its usefulness with gel and film product formulations. Methods: Study arms of four to 24 female macaques completed each post-coital safety experiment. Formulations tested include two placebo films (polyvinyl alcohol and cellulose based platforms; n=4 each), one active gel (reduced glycerin tenofovir; n=8), one placebo gel (HEC; n=24), and no product (n=24). Experiment days and 3 consisted of two vaginal exams (Time-0 and Time-30 minutes, in reference to product application), with no opportunity for coital activity. On days 2 and 4, each female macaque was housed with a male macaque for 5minutes shortly after vaginal sampling and intravaginal product administration. Females were sedated for post-coital exam including colposcopy, vaginal flora, ph and smear. Days 5 and 8 each consisted of a single vaginal exam. Safety measures include colposcopy to monitor the mucosal integrity of the cervicovaginal tissues, assessments for vaginal microflora fluctuations, notations of vaginal ph, and Gram stain analysis of vaginal smears to detect inflammatory response by PMN infiltration. Results: Coitus increases cervicovaginal erythema, petechiae and ecchymosis notations at colposcopy. None of the tested formulations exacerbated these or induced other colposcopic findings. Vaginal ph generally decreased after exposure to formulations other than HEC gel, but increased when semen was present after coitus. Vaginal flora and PMN presence were not altered by the formulations tested. Conclusions: These experiments indicate that vaginal gel and film formulations can be assessed in the macaque post-coital safety model. Additionally, the reduced glycerin formulation of tenofovir gel is shown to be safe by parameters assessed in this model. 90 HIV Research for Prevention 204 HIV R4P

91 Wednesday, 29 October Oral Abstract Session 9: Good Participatory Practices in HIV Prevention OA9.0 OA9.02 Inclusion of Transgender and Gender Nonconforming Communities in Preventive HIV Vaccine Research at the HIV Vaccine Trials Network (HVTN) Gail B. Broder,2, Michele P. Andrasik,3, Shelly T. Karuna,3, Transgender Working Group of the NIAID-funded HIV Vaccine Trials Network Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 HIV Vaccine Trials Network, Communications and Community Engagement Units, Seattle, WA, United States, 3 HIV Vaccine Trials Network, Clinical Development Unit, Seattle, WA, United States Background: The HVTN has developed practices to increase representation of transgender and gender non-conforming (trans*) communities in its clinical trials since Recognizing that this population has elevated HIV incidence, the HVTN has undertaken ongoing efforts to engage and include this population in preventive vaccine trials so that they contribute to finding a vaccine for use by those who are most in need. Methods: A multidisciplinary Transgender Working Group was formed in The group has reviewed and revised data collection forms and protocol template language; developed trainings to improve cultural responsiveness; and provided a forum for development of research proposals to better understand and serve trans* participants. Results: This focus on the inclusion of trans* people has led to several HVTN policy and practice changes. Demographics forms collect data on sex at birth and gender identity as 2 independent variables. Protocols require pregnancy testing and use of birth control only by those who are biologically capable of bearing children. HVTN505 was HVTN s first HIV vaccine study to specify eligibility of male-to-female transwomen (not grouped with MSM). Focus groups were conducted with transwomen to collect information about barriers and facilitators to enrollment in HIV vaccine trials. Ongoing analysis of trans* participants in HVTN trials continues to increase understanding. Conclusions: While sexual orientation has become a standard part of demographics vocabulary in clinical trials, gender identity remains poorly understood. Laboratory reference ranges and immunogenicity analyses are often based on birth sex, thus complicating the interpretation of laboratory data from trans* participants. Culturally responsive trial conduct is enhanced by consultation with community stakeholders and by attention to the needs and experiences of trans* people in clinical and outreach settings. Collecting and sharing data regarding trans* people in clinical HIV prevention trials is critical. Breaking Barriers: Do Research Interests Meet the Needs of the Lesbian, Gay, Bisexual and Transgender (LGBT) Community in Kenya? Jonah M. Chinga, George V. Owino 2, Kent Klindera 3, Prince N. Bahati 4 Minority Persons Empowerment Group (MPEG), Programs, Thika, Kenya, 2 International AIDS Vaccine Initiative (IAVI), External Relations, Nairobi, Kenya, 3 amfar, GMT, New York, NY, United States, 4 International AIDS Vaccine Initiative (IAVI), Research Preparedness - External Relations, Nairobi, Kenya Background: LGBT community in Kenya continues to be criminalized under the penal code. Research with LGBT individuals in the context of dynamic human rights and heightened criminalization requires higher scrutiny of ethical guidance and community ownership. The LGBT and research community in Kenya aimed to develop partnership for common research agenda and ethical guidelines. Methods: A two-day participatory workshop with 24 LGBT leaders with experience with research was convened to identify current ethical challenges, solutions, HIV and Sexual Orientation and Gender Identity (SOGI) related research priorities in Kenya. Results: A mismatch between researchers priorities and community needs has led to inadequate community consultations. A complex informed consent process, misdirected motivation for participation (e.g. transport reimbursement and health care) was identified as the top ethical challenges. HIV research priorities identified by LGBT included the need to demonstrate the link between homophobia, transphobia and HIV; factors affecting adherence; impact of gender-based violence on HIV infection among LGBT. Other areas included effects of drug and substance use, investigation of causes behind the higher incidence rate among MSM sex workers compared to female sex workers practicing anal sex. They recommended mainstreaming of SOGI and human rights issues in HIV research; studying the impact of the heightened criminalization on coming out and on participation in research. A national representative LGBT research advisory group was formed to advise, guide and facilitate linkages between researchers and LGBT community. Conclusions: There is need for greater involvement of LGBT community in all stages of research. Strengthening the capacity of LGBT organizations is instrumental in ensuring ethical conduct of HIV research in rights constrained settings. Sustainable partnership lies in matching researchers agenda with community research needs and interests. ORAL ABSTRACT SESSIONS 9

92 Oral Abstract Sessions Oral Abstract Session 9: Good Participatory Practices in HIV Prevention OA9.03 OA9.04 ORAL ABSTRACT SESSIONS Words & Images Have Consequences: Gay Men, Prevention Research and the Doubleedged Sword of the Media Cindra Feuer, Michael Ighodaro 2, Brian Kanyemba 3, Kay Marshall, James Pickett 4, Paul Semugoma 5, Maaza Seyoum 6 AVAC, New York, NY, United States, 2 IGLHRC, New York, NY, United States, 3 Desmond Tutu HIV Foundation, Cape Town, South Africa, 4 IRMA, Chicago, IL, United States, 5 ANOVA Health Institute, Health4Men Programme, Cape Town, South Africa, 6 IAVI, Johannesburg, South Africa Background: Homophobia, stigma and criminalization hamper efforts to include MSM in HIV research and programming in Africa when gay men and other MSM are at high risk for HIV. Trial sites and community advocates must consider how to best make use of media to humanize MSM and support research while protecting individual confidentiality and safety and the stability of trial sites, especially as new harsh anti-gay laws are passed. Methods: After a Washington Post article quoted gay Africans and forced a Nigerian man to seek asylum in the US advocates were inspired to develop media engagement best practices. At ICASA 203, AVAC and IRMA convened two sessions for MSM to collect strategies to maximize benefits and minimize harms when engaging media. Conversations with journalists explored strategies for reporting on MSM. We are documenting tactics to share with researchers, advocates and media. Results: Participants discussed courting journalists and harnessing personal stories to gain media attention while protecting identities. They noted the need to balance safety with visibility, and strategized how to do both. Media was seen as critical to counteract homophobia with public health messages for inclusion of MSM in HIV research and services. Yet even positive stories that include names or other identifying details can put individuals and organizations at risk. Experienced advocates shared methods to respond to and avoid crises, including the use of media generated by MSM. Journalists noted the importance of educating editors. Conclusions: Best practices for garnering media coverage that is sensitive, safe, accurate and benefits vulnerable groups in HIV research should be shared with advocates and researchers, particularly in criminalized settings. Communication plans particular to MSM will assure recruitment, retention and protection of research participants and integrity of research sites. Education of journalists about needs of vulnerable populations and benefits of research programs should be a priority. The Utilization of Good Participatory Practice (GPP) during the Planning and Implementation of a PrEP Study among Black MSM Jonathan Paul Lucas, Georgette King, Phaedrea Watkins, Christopher Chauncey Watson 2, Craig S. Hutchinson 3, Christine Rogers, S Wakefield 4, Sheldon D. Fields 5 FHI 360, Science Facilitation, Durham, NC, United States, 2 The George Washington University, Washington, DC, United States, 3 UCLA, Semel Institute for Neuroscience & Human Behavior, Los Angeles, CA, United States, 4 HIV Vaccine Trials Network, Seattle, WA, United States, 5 Florida International University, Nicole Wertheim College of Nursing & Health Sciences, Miami, FL, United States Background: Black men who have sex with men (BMSM) in the United States (US) are disproportionately impacted by HIV despite having no greater risk profile than their White counterparts. Deficits in AIDS/HIV knowledge and distrust in research among BMSM have been cited as common barriers to vital research engagement. To address potential concerns and build community rapport, community engagement planning for HPTN 073, a Demonstration project designed to assess the initiation and correlates of daily pre-exposure prophylaxis (PrEP) use by BMSM, required innovative approaches to ensure robust community involvement during the study design and pre-implementation process. Methods: In 203 the HPTN 073 Community Working Group utilized GPP guidelines to conduct three half-day community consultations with BMSM service providers, community based organizations and stakeholders in Washington, DC; Chapel Hill, North Carolina; and Los Angeles, California. Consultation attendees (n=38) were provided overviews of global PrEP research and the HIV epidemic among BMSM in the US, as well as a description of HPTN 073. Participants provided feedback on HPTN 073 study design and implementation plans. Results: Feedback from the community consultations resulted in alterations to the HPTN 073 social marketing campaign. The changes were incorporated to support community engagement, encourage study enrollment and increase BMSM PrEP awareness. Cultural competency training for site staff and healthcare providers was added and modifications were made to HPTN 073 s participant recruitment, retention and adherence plans. Conclusions: Engaging community members in problem-solving solutions to issues that affect them is one of the fundamental principles of HIV prevention research. It is a critical strategy for partnership building to advance HIV prevention efforts, particularly with marginalized groups. Effective utilization of GPP can aid in the development of shared responsibility and community ownership of HIV prevention research initiatives. 92 HIV Research for Prevention 204 HIV R4P

93 Wednesday, 29 October Oral Abstract Session 9: Good Participatory Practices in HIV Prevention OA9.05 OA9.06 Assessing GPP in Action: How FACTS 00 Formalised the Implementation of Stakeholder Engagement in a Large-scale Prevention Trial Sinazo Pato, Deborah Baron, Busi Nkala 2, Phumeza Mzizi 2, John Mdluli 3, Ireen Mosewu 4, Leader Kanyiki 5, Duduzile Lembethe 6, Zonke Mabude 6, Ronald Moate 7, Isaac Ngema 8, Ross Malamatsho 9, Cynthia Dlamini, Sinead Delany-Moretlwe, Glenda Gray 2, Helen Rees Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 2 Perinatal HIV Research Unit, Soweto, South Africa, 3 The Aurum Institute, Johannesburg, South Africa, 4 The Aurum Institute, Rustenburg, South Africa, 5 Desmond Tutu HIV Foundation, Cape Town, South Africa, 6 MatCH Research, Pietermaritzburg, South Africa, 7 Medunsa Clinical Research Unit, GaRankua, South Africa, 8 Qhakaza Mbokodo Research Clinic, Ladysmith, South Africa, 9 Setshaba Research Centre, Soshanguve, South Africa Background: The Good Participatory Practice (GPP) Guidelines and Tools for Biomedical HIV Prevention Trials aim to ensure effective stakeholder engagement throughout trials. GPP Guidelines (2 nd edition) have been published, but experience with translation from principles to practice is limited. We evaluated the formal implementation of 6 GPP principles within the FACTS 00 trial, a phase III licensure trial of tenofovir % gel conducted across 9 sites in South Africa. Methods: Prior to trial initiation, staff and community advisory boards (CABs) at all sites received GPP training. Sites subsequently produced comprehensive GPP plans; these were reviewed and updated quarterly, and sites reported monthly on the implementation of the plans. We evaluated 44 GPP plans and 29 monthly reports produced by sites between January 202 and July 203 to determine the extent to which the GPP principles were applied and found effective. Results: FACTS sites effectively applied all of the GPP principles relevant to sites before and during implementation of the trial. This was achieved through support from a full-time GPP manager based in the CORE team, formal incorporation of GPP procedures into the Manual of Procedures, extensive staff and CAB trainings, and regular review of site materials. While plans were not always implemented as designed, sites developed site-specific strategies and used new tools to meet specific needs. Challenges included staff turn-over at sites, and the need for repeat training in GPP planning and reporting. Balancing the requirement for multiple tools to monitor GPP formally, without making monitoring burdensome, was a key lesson learned. Conclusions: The GPP guidelines provided a framework for the development of effective stakeholder engagement in the FACTS trial. Site-specific GPP plans provide useful over-arching guidance and strategies needed for rapid response situations. Challenges with Participant Reimbursement: Experiences from CAPRISA A Post-trial Access Study Kathryn T. Mngadi, Janet Frohlich, Carl Montague, Jerome Singh,2,3, Nelisiwe Nkomonde, Nomzamo Mvandaba, Fanelesibonge Ntombela, Londiwe Luthuli, Quarraisha Abdool Karim,4, Leila Mansoor Centre for the Programme of AIDS Research in South Africa, HIV Prevention Research, Durban, South Africa, 2 Sandra Rotman Centre, University Health Network, Toronto, ON, Canada, 3 Dalla Lana School of Public Health and Joint Centre for Bioethics, University of Toronto, Toronto, ON, Canada, 4 Mailman School of Public Health, Columbia University, Department of Epidemiology, New York, NY, United States Background: Reimbursement of trial participants is a controversial issue, with post-trial access studies necessitating new strategies and models. CAPRISA 008, a post-trial access study testing the feasibility of using family planning services to rollout a pre-licensure HIV prevention intervention (% tenofovir gel), tried to balance the real life scenario of no reimbursement for attendance at public sector clinics, with recognition of the burden borne by participants in the preceding efficacy trial, by offering minimal reimbursement for study visits. This reimbursement was meant to subsidise transport to the clinic, but impacted negatively on accrual, retention and participant morale. Methods: A review of relevant local and international guidelines was conducted by CAPRISA s bioethicist, as were consultations between the senior protocol team, participants and the community. Subsequently the institutional policy on reimbursement was revised, as per guideline recommendations. A revised reimbursement schedule for this study was submitted to, and approved by the University of Kwazulu-Natal s Biomedical Research Ethics Committee (UKZN BREC). Results: The review of guidelines recommended compensation for participant time, travel and inconvenience. The retention rate for December 202 was 63.0% (standard 90%); UKZN BREC approval of the revised rate occurred in February 203, and retention in March 203 rose to 92.2%. To date the overall retention is 94.6% (March 204). Conclusions: Changes in reimbursement schedules between efficacy and post-access trials, without prior consultation with participants and community representatives, may negatively impact accrual and retention, with the potential to reduce sample size and impair the validity of the findings. The question remains if community members who did not participate in the efficacy study, should receive the same (if any) reimbursement as the original participants, if included in post-trial access studies. ORAL ABSTRACT SESSIONS 93

94 Oral Abstract Sessions Oral Abstract Session 20: Reproductive Hormones and HIV Risk OA20.0 Medroxyprogesterone Acetate Enhances HIV- Uptake and Transcytosis, but not Replication, in Primary Human Genital Epithelial Cells Victor H. Ferreira, Aisha Nazli, Jessica K. Kafka, Kristen Mueller, Michel J. Tremblay 2, Alan Cochrane 3, Charu Kaushic McMaster University, Hamilton, ON, Canada, 2 Laval University, Laurier, QC, Canada, 3 University of Toronto, Toronto, ON, Canada OA20.02 LB The Contraceptive MPA, Unlike NET, Modulates Expression of Immune Function Genes and Increases HIV- Infection in Cervical Tissue Explants and PBMCs Roslyn M. Ray, Chanel Avenant, Johnson M. Moliki, Janet P. Hapgood The University of Cape Town, Molecular and Cell Biology, Cape Town, South Africa ORAL ABSTRACT SESSIONS Background: Half of all people living with HIV worldwide are women, yet early events in the female genital tract are poorly understood particularly in the context of female sex hormones or hormonal contraceptives, such as medoxyprogesterone acetate (MPA), which have been correlated with increased HIV susceptibility and transmission. Methods: Genital epithelial cell (GEC) cultures were prepared from human tissues and grown in the presence or absence of physiological concentrations of estrogen (E2), progesterone (P4) or MPA prior to HIV- exposure. HIV uptake was measured by p24 ELISA in disrupted GECs and localization of virus was confirmed by electron microscopy (EM). Various inhibitors were used to determine the pathways involved in viral uptake. HIV infection and replication were determined by using highly sensitive real time PCR assays. Apical recycling and transcytosis were assessed by measuring p24 antigen in apical or basolateral GEC supernatants, respectively, and infectiousness of the virus was determined by TZM-b assay. Results: HIV uptake was significantly increased within endometrial and cervical GECs grown in the presence of MPA. HIV- uptake by GECs primarily took place via endocytosis, since treatment with Dynasore, an endocytosis inhibitor, significantly decreased HIV uptake. EM confirmed that HIV- was localized to intracellular vesicular compartments. Despite uptake into GECs, no early or late reverse transcription products, integrated HIV DNA or spliced RNA transcripts were measured, regardless of hormone exposure. Nevertheless, HIV- transcytosis was significantly increased among GECs grown in the presence of P4 and MPA and this virus was infectious. Conclusions: These results suggest that female sex hormones, particularly MPA, regulate HIV transcytosis across the epithelium and HIV uptake into GECs, but not replication. Ongoing studies are investigating whether enhanced transcytosis in the absence of productive infection, would result in increased HIV transmission to target cells. Background: The synthetic progestins, medroxyprogesterone acetate (MPA) and norethisterone enanthate (NET-EN), are widely used in developing countries as injectable contraceptives, where disease burden is high. Some studies suggest that MPA, unlike NET, increases HIV- acquisition in women. Whether MPA and NET differentially affect HIV- infection and the expression of key genes relevant to HIV- acquisition via differential molecular mechanisms, is key to understanding choice of progestin contraceptive for HIV- prevention. Methods: Regulation of selected genes was investigated in cervical tissue explants and peripheral blood mononuclear cells (PBMCs) by qrt- PCR, western blotting and Luminex assays, in response to physiologically relevant doses of progestogens. Infection assays were performed in the absence and presence of HIV- using HIV- BAL-RENILLA or HIV pnl4.3 IMCs. The GR specific antagonist RU486 or GR sirna knockdown were used to determine the role of the GR in modulating ligand-specific effects. Results: In PBMCs, MPA like dexamethasone (DEX, a GR specific agonist), showed anti-inflammatory effects, decreasing pro-inflammatory IL6, IL8 and RANTES levels and increasing anti-inflammatory GILZ gene expression levels, while NET and progesterone (P4) did not. In primary cervical tissue explants, DEX and MPA repressed IL6 and IL8 and increased GILZ gene expression levels. Differential gene expression by MPA versus NET and P4 were mediated via the GR in PBMCs. Similarly, MPA and DEX, unlike NET and P4, increased HIV- replication in viable PBMCs. In primary cervical explants, MPA, but not NET increased HIV- replication. Conclusions: Collectively, the data suggest that NET, unlike MPA, would be a safer choice of injectable progestin contraceptive in young women in high risk areas for HIV- infection. The molecular basis for this choice most likely involves differential effects of MPA as compared to NET and P4, on transcription of immunomodulatory genes, due to their differential actions via the ubiquitous GR. 94 HIV Research for Prevention 204 HIV R4P

95 Oral Abstract Session 20: Reproductive Hormones and HIV Risk Wednesday, 29 October OA20.03 Hormonal Regulation of Early Innate Immune Responses to HIV Infection by Dendritic Cells from the Human Female Reproductive Tract Marta Rodriguez-Garcia, Zheng Shen, John V. Fahey, Austin W. Boesch 2, Margaret E. Ackerman 2,3, Charles R. Wira Geisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United States, 2 Thayer School of Engineering, Hanover, NH, United States, 3 Geisel School of Medicine at Dartmouth, Microbiology and Immunology, Hanover, NH, United States OA20.04 Injectable Contraceptive Use Correlates with Increased HIV Target Cells at the Cervix in Young South African Women Elizabeth H. Byrne, Melis Anahtar, Kathleen Doherty,2, Gregory Olson, Brittany Bowman, Nikita Padavattan 3, Zaza Ndhlovu,3, Musie Ghebremichael, Bruce Walker, Thumbi Ndung u,3, Krista Dong, Douglas S. Kwon Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 Vanderbilt University School of Medicine, Nashville, TN, United States, 3 HIV Pathogenesis Programme, Durban, South Africa Background: Dendritic cells (DC) in tissues differ considerably from classical in vitro models. DCs are key for capturing and transferring HIV to CD4+ T cells as well as inducing adaptive immune responses. A major gap remains in our knowledge regarding phenotype, innate responses and regulation of DCs in the female reproductive tract (FRT), one of the main portals of entry for HIV. Here we investigated hormonal regulation of DC from different anatomical sites in the FRT and their early innate responses to HIV. Methods: Tissues from the endometrium (EM), endocervix (CX) and ectocervix (ECX) were digested prior to DC purification using CD4 and CDa magnetic bead selection. Cells were analyzed by flow cytometry or stimulated with HIV for 3 and 24 hr after estradiol (E 2 ) treatment (5x0-8 M). Supernatants were analyzed for 9 different antimicrobials with anti-hiv activity, using a multiplex assay developed by us. Results: We identified distinct populations of DC/macrophages in the FRT, based on their expression of CDc and CDb. Population distribution was different between tissues, with DCs more abundant in EM and ECX than CX. Higher expression of HLA-DR was detected in DC from the ECX, but no differences were found in CD4 and CD03 expression between tissues. Analysis of supernatants revealed that CDa+ and CD4+ cells constitutively produced HNP-3, SLIP, RANTES, Elafin, Lactoferrin, and MIP3alpha. Treatment with E 2 for 24h increased production of HNP-3, RANTES, MIP3alpha and SLPI. HIV stimulation induced HNP-3, RANTES and Elafin within 3hr, but interestingly when DC were pretreated with E 2, the stimulatory effect of HIV was undectable. Conclusions: Different DC populations exist in the FRT that may play different roles in HIV pathogenesis. DC produce antimicrobials shortly after HIV challenge and these responses are modified by E 2. Overall, our results indicate hormonal regulation of innate immune responses by DC to HIV, which may have consequences for alterations in HIV susceptibility in the FRT. Background: The use of progestin-only injectable contraception (IC) has been epidemiologically associated with increased risk of HIV acquisition, but it remains unclear whether this relationship is causal and, if so, the mechanisms by which progestin-only contraceptives increase risk. Methods: Demographic and behavioral data along with blood and female genital tract (FGT) samples were obtained from a cohort of 8-23 year old HIV uninfected women in Durban, South Africa. Cervical cytobrush cells were analyzed by flow cytometry and plasma progesterone measured by chemiluminescence. Cytokines in cervicovaginal lavage (CVL) were measured by luminex. Results: Women using IC disproportionately accounted for new HIV infections during the study (RR 2.55 [ ], p=0.008). While 3.5% of women in the cohort used IC, 72.7% of women who became HIV positive used IC (8 of ); 9.09% of infected women did not use contraception. This difference was not accounted for by behavioral or demographic differences. The use of IC was significantly associated with a higher frequency of CD4+CCR5+ T cells of CD45+ cells in the cervix relative to those not on hormonal contraception (.8% [0.33, 4.53] vs. 0.29% [0.3, 0.48], respectively; p=0.036). Women with high endogenous progesterone (>0.3ng/mL, luteal phase) in the absence of IC use had significantly increased frequency of CD4+CCR5+ T cells in the cervix relative to those with low progesterone (follicular phase;.6% [0.5, 3.76] of CD45+ cells vs 0.29% [0.3, 0.48], respectively; p=0.042). High endogenous progesterone was also associated with a higher frequency of cervical CD4+CCR5+HLADR+CD38+ T cells (p=0.034). There was no observed difference based on IC use in soluble cytokine or chemokine levels in CVL. Conclusions: The increased frequency of HIV target cells at the site of exposure in the context of IC and high endogenous progesterone provides a potential biological mechanism for the increased HIV acquisition risk for women using injectable progestin-only contraceptives. ORAL ABSTRACT SESSIONS 95

96 Oral Abstract Sessions Oral Abstract Session 20: Reproductive Hormones and HIV Risk OA20.05 LB OA20.06 ORAL ABSTRACT SESSIONS Increased Number of HIV Target Cells in Zimbabwean vs US Women Sharon L. Achilles,2, Felix G. Mhlanga 3, Allen T. Matubu 3, Kevin A. Stoner 2, May A. Beamer 2, Z. Mike Chirenje 3, Sharon L. Hillier,2 University of Pittsburgh, Obstetrics, Gynecology and Reproductive Sciences, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 3 UZ - UCSF, Harare, Zimbabwe Background: Hormonal contraception (HC), especially depot medroxyprogesterone acetate (DMPA), has been associated with increased HIV risk. Several investigators are evaluating the impact of HC on immune cells and immune mediators in the female genital tract (FGT). Understanding baseline differences in these immune factors in different populations is critical to the evaluation of the impact of HC. Our objective was to compare immune cells from a Sub-Saharan African (SSA) population of women and a North American (US) population of women using identical sampling and analytical methods. Methods: Women in Pittsburgh, PA (US) and in Harare, Zimbabwe (SSA) aged 8-34 who had not used hormonal or intrauterine contraception for >30 days and DMPA for >0 months were enrolled into a study designed to quantify systemic and FGT immune cell populations. Enrolled women screened negative for HIV, gonorrhea, chlamydia, trichomonas and active herpes. Immune cell populations collected from blood and endocervical cytobrushes were quantified by flow cytometry. All flow cytometric data were gated in a single lab. Results: Eighty US women and 80 SSA women were enrolled. SSA women had a higher mean number of CD4 cells 3443 vs 674, p < 0.00), CD4CCR5+ cells (696 vs 887, p=0.002), and activated CD4 cells (expressing CD69) (2066 vs 233, p< 0.02) in the cervix compared to US women. SSA women also had more CD4CCR5+ cells (3775 vs. 547, p< 0.00) and CD4CCR6+ cells (,053 vs. 3869, p< 0.00), and had fewer activated CD4 cells (expressing CD69) (2085 vs. 295, p< 0.0) in the PBMCs compared to US women. Conclusions: There are significant differences in the numbers and activation status of both peripheral and local CD4 cells in US and SSA women, with SSA women having both greater mean CD4 and CCR5 expressing T-cells compared to US women. Studies evaluating the impact of HC on FGT immune cells should consider baseline cellular populations. HC induced changes in immune cells may be mitigated by the number and type of cells at baseline. Hormonal Contraception Use and Risk of Female-to-Male HIV Transmission Risk in a Zambian Cohort Kristin M. Wall,2, William Kilembe 3, Htee Khu Naw 3, Ilene Brill 3, Bellington Vwalika 3, Elwyn Chomba 3, Brent Johnson, Lisa Haddad, Amanda Tichacek 2, Susan Allen 2 Emory University, Atlanta, GA, United States, 2 Rwanda Zambia HIV Research Group, Atlanta, GA, United States, 3 Rwanda Zambia HIV Research Group, Lusaka, Zambia Background: The association between hormonal contraception (HC) use and risk of both male-to-female and female-to-male transmission HIV transmission is highly debated. Very little data exists related to femaleto-male transmission risk and HC use. USAID, FHI360, and the World Health Organization have called for more evaluations of this association. Methods: HIV discordant couples in which the man was negative and the woman was positive (M-F+) were identified after couples voluntary HIV counseling and testing from in Lusaka, Zambia. Discordant couples were followed longitudinally and demographic, behavioral, clinical, and family planning measures were collected at baseline and 3-monthly. Men were re-tested for HIV every three months. Multivariate Cox models evaluated time to HIV acquisition among men. Results: Among 654 M-F+ couples, 226 incident infections occurred over 3366 couple-years (6.7/00 couple-years; 95%CI: ). 7 (76%) infections were genetically linked to the study partner. No interaction between genital ulceration/inflammation and contraception was observed. Use of injectables (HR=0.7; 95%CI:0.4-.4), OCPs (HR=.3; 95%CI:0.7-2.), or implants (HR=0.8; 95%CI: ) in the past three months was not associated with genetically linked HIV transmission from women to men relative to non-hc controlling for: woman s age, number of previous pregnancies, woman s log viral load at baseline, male circumcision status, pregnancy status, sex frequency with and without a condom, sperm on a wet mount, male and female genital inflammation and ulceration, and time interval since enrollment. Conclusions: Our results add to a small and inconclusive body of literature. Over 7 years of follow-up, we found no statistically significant association between HC use and female-to-male HIV transmission. These findings support the continued use of HC methods for pregnancy prevention and Prong 2 of PMTCT. 96 HIV Research for Prevention 204 HIV R4P

97 Oral Abstract Session 2: Viral Transmission Studies Thursday, 30 October OA2.0 Transmission of Pre-adapted Viruses Determines the Rate of CD4 Decline in Seroconverters from Zambia Daniela Monaco, Dario Dilernia, Malinda Schaeffer, Kristine Dennis, Jonathan Carlson 2, Jessica Prince, Daniel Claiborne, William Kilembe 3, Shabir Lakhi 3, James Tang 4, Matt Price 5, Paul Farmer, Richard Kaslow 4, Jill Gilmour 6, Susan Allen, Paul Goepfert 4, David Heckerman 2, Eric Hunter Emory University, Atlanta, GA, United States, 2 Microsoft Research, Redmond, WA, United States, 3 Zambia Emory HIV Research Project, Lusaka, Zambia, 4 University of Alabama-Birmingham, Birmingham, AL, United States, 5 IAVI, San Francisco, CA, United States, 6 IAVI, London, United Kingdom OA2.02 The Sequence of the α4β7-binding Motif on Gp20 of Transmitted/Founder Viruses Contributes to the Dependence on the Integrin for HIV Infection Simone I. Richardson,2, Elin Gray, Nonhlanhla Mkhize,2, Daniel Sheward 3, Bronwen Lambson,2, Kurt Wibmer,2, Lindi Masson 3, Lise Werner 4, Nigel Garett 4, Jo-Ann Passmore 3, Salim Abdool- Karim 4, Carolyn Williamson 3, Penny Moore,2, Lynn Morris,2 Centre for HIV and STI s, National Institute for Communicable Diseases, Johannesburg, South Africa, 2 School of Pathology, University of the Witwatersrand, Johannesburg, South Africa, 3 Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 4 Centre for the AIDS Programme of Research in South Africa (CAPRISA), Nelson R Mandela School of Medicine, University of KwaZulu-Natal, Durban, South Africa Background: HIV escapes adaptive cellular immunity by selecting mutations that are associated with the individual s HLA-I alleles. These mutations can be transmitted but the impact of this process on pathogenesis is poorly understood. Methods: In 69 transmission pairs, we studied the transmission of HIV polymorphisms in Gag, Pol and Nef by Sanger sequencing of population amplicons in the donor (D) and the linked-recipient (LR) ( 3 months post-transmission). Polymorphisms statistically-linked to HLA alleles or located in well-defined CTL epitopes were quantified according to each LR s HLA alleles and associated with their set-point VL and CD4 counts. Results: The majority of polymorphisms (83.6%) were transmitted from the D to the LR and a significant fraction (7.3%) was already adapted to the LR s HLA (.6% escape and 6.2% epitope-located). A Spearman correlation analysis showed that transmission of Pol polymorphisms irrelevant to the LR s HLA was associated with a diminished set-point VL (p=0.003). This association was lost (p=0.4) when other variables known to determine set-point VL (gender-p=0.0; B*57-p=0.02; HLA-B sharing-p=0.006; replicative capacity (RC)-p=0.008) were included in a Generalized Linear Model. An in-depth analysis of survival curves (log-rank test) for different CD4 endpoints ( cells/ul) showed that the proportion of transmitted HLA-linked polymorphisms relevant to the LR HLA in Gag was consistently associated with a faster CD4 decline (p=0.0004). When other factors (gender, protective alleles, allele sharing, RC and set-point VL) were considered in a Cox Proportional Hazard Model, the proportion of transmitted HLA-linked polymorphisms in Gag remained the only variable significantly associated with CD4 decline (p=0.03). Conclusions: Because most Gag, Pol and Nef polymorphisms are transmitted, newly infected individuals can receive a pre-adapted variant that leads to an accelerated disease progression (faster CD4 decline) without showing a significant effect on set-point VL. Background: The integrin α4β7, which mediates the trafficking of T lymphocytes to the gut associated lymphoid tissue (GALT), a site of rapid HIV replication, has been described as an attachment factor for the V2 loop of the envelope protein gp20. We aimed to study the factors that influence dependence on α4β7 for replication of transmitted/founder viruses including cytokine levels in cervicovaginal lavage (CVL), STI infections and the sequence of the tripeptide α4β7-binding motif. Methods: All-trans retinoic acid-activated CD4+ T cells were incubated with or without HP2/ (anti-α4 antibody) or Act- (anti-α4β7 antibody) prior to adding virus. Infectious virus was prepared using envelope genes of the transmitted/founder (T/F) virus from 8 individuals in the CAPRISA Acute Infection cohort. Replication was monitored by p24 ELISA. Changes in viral sequence were generated by site-directed mutagenesis. Results: T/F viruses with the highest dependence on α4β7 for replication had P/SDI/V motifs while those with lower dependence were LDI/L. Mutation of viruses with LDI/L motifs to P/SDI/V resulted in increased dependence on α4β7 for replication while the reverse mutation restricted the ability of the viruses to enter cells. T/F viruses from individuals diagnosed with bacterial vaginosis (BV) at the time of virus isolation had significantly higher dependence on the integrin for replication. Levels of IL-7, a cytokine that upregulates α4β7 expression, correlated with α4β7 dependence in the CVL shortly after transmission. Both BV status and high IL-7 levels in the CVL were associated with the P/SDI/V motifs in a larger cohort of 28 CAPRISA 002 participants. Conclusions: P/SDI/V motifs are more common among South African HIV subtype C viruses accounting for 35% of variants. These data suggest that viruses with P/SDI/V motifs favour α4β7 reactivity at transmission influenced by the presence of BV and IL-7 cytokine levels. These findings may lead to vaccine and therapeutic opportunities in which α4β7 reactivity is exploited. ORAL ABSTRACT SESSIONS 97

98 Oral Abstract Sessions Oral Abstract Session 2: Viral Transmission Studies OA2.03 OA2.04 ORAL ABSTRACT SESSIONS HIV Replicative Capacity of Transmitted Viruses Is Associated with Early Immune Activation, Exhaustion and Establishment of the Viral Reservoir Jessica L. Prince, Daniel T. Claiborne, Gladys Macharia 2, Luca Micci, Benton Lawson, Eileen Scully 3, Jakub Kopycinski 4, Thomas Vanderford, Jianming Tang 5, Tianwei Yu, Shabir Lakhi 6, William Kilembe 6, Guido Silvestri, Paul Goepfert 5, Matthew A. Price 2,7, Marcus Altfeld 8, Mirko Paiardini, Jill Gilmour 2, Susan Allen,6, Eric Hunter Emory University, Atlanta, GA, United States, 2 International AIDS Vaccine Initiative (IAVI), London, United Kingdom, 3 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States, 4 Imperial College London, London, United Kingdom, 5 University of Alabama at Birmingham, Birmingham, AL, United States, 6 Zambia Emory HIV Research Project, Lusaka, Zambia, 7 UCSF, San Francisco, CA, United States, 8 Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany Background: Determining the host and viral factors that shape the trajectory of early HIV- pathogenesis is key for developing rational prevention strategies. Previously, we showed that in individuals recently infected with HIV- subtype C, low viral replicative capacity (RC) as defined by the transmitted Gag sequence, was associated with a delayed loss of CD4 T cells independent of set point VL and host immunogenetic factors. We hypothesize that low RC leads to a muted inflammatory response characterized by reduced immune activation, might attenuate infection of memory T cell subsets and preserve critical CD4 T cell homeostasis. Methods: Levels of plasma cytokines at seroconversion were measured using a Luminex platform. Flow cytometry was used to assess markers of activation (CD38+HLADR+), exhaustion (PD- and CD57), and proliferation (Ki-67) on CD4 and CD8 cells. Cell associated viral DNA in CD4 memory populations was quantified with qpcr. Results: RC was positively correlated with levels of inflammatory cytokines in plasma and with activation of both CD8 (p=0.0) and central memory (CM) CD4 T cells (p=0.002). Low RC was associated with CD8 T cells that were less exhausted (p< 0.00) and more cytotoxic (p=0.002). RC was positively correlated with proliferation (p=0.003) and with the level of cell associated viral DNA in CM CD4 T cells (p=0.0), a population highlighted to be integral for the maintenance of latency and preferentially spared in non-pathogenic SIV infection. Consistent with previous studies, we observed that cellular immune activation, proliferation, exhaustion, and cell associated viral DNA in CM CD4 T cells were all associated with the rate of disease progression. Conclusions: This study highlights the integral role that RC of the transmitted virus plays in defining several facets of HIV- immunopathology. Understanding the complex interactions between HIV and the immune system will be crucial for designing innovative prevention strategies. In-vitro Fitness of HIV- Transmitted/Founder versus Non-transmitted Full-length Genome Infectious Molecular Clones Martin J. Deymier, Zachary Ende, Daniel T. Claiborne, William Kilembe 2, Susan Allen,2, Eric Hunter,2 Emory University, Atlanta, GA, United States, 2 Zambia Emory HIV Research Program, Lusaka, Zambia Background: In ~80% of heterosexual transmissions of HIV-, an infected individual with a diverse viral quasispecies transmits a single viral variant, the Transmitted/Founder (TF), to a naïve host. Evidence is building that TF variants are enriched for certain genetic and phenotypic characteristics that presumably enhance the efficiency of transmission. However, the mechanisms involved are largely ambiguous, partially because studies using full-length genomes in transmission pairs are lacking. Methods: We have performed HIV near full-length (NFL) single genome amplification from six subtype C acutely infected individuals and each of their chronically infected virologically linked partners in the Zambia- Emory HIV Research Project. Phylogenetic analysis performed on the 8 NFL genomes (mean 8/transmission pair) confirms epidemiologically linked transmission as well as infection by a single viral variant in each case. We have generated 5 TF & 34 non-transmitted (NT) full-length infectious molecular clones from 5 transmission pairs and assayed for particle infectivity by dividing the virus titer on TZM-bl cells by the RT activity of the virus stock. Results: The particle infectivity of the TF compared to the median of the NT variants for all matched transmission pairs was not statistically significant (p=0.22). However, particle infectivity correlated with the amount of glycosylation on the Env V-V4 region (R=0.40, p= 0.0) as well as with replication in PBMCs for a subset of tested viruses (R=.823 p=0.0), suggesting that previous findings showing less glycosylation on TF viruses could mean lower replicative capacities in vitro. However, preliminary data suggests that lower replicating, less glycosylated viruses, may preferentially productively infect monocyte-derived dendritic cells. Conclusions: Understanding the characteristics of TF viruses that allow for efficient transmission will aid in prophylaxis and early intervention efforts. JP and DC contributed equally to this work 98 HIV Research for Prevention 204 HIV R4P

99 Oral Abstract Session 2: Viral Transmission Studies Thursday, 30 October OA2.05 Genetic Footprints within the HIV- Envelope Glycoprotein Associated with Transmission in Men who Have Sex with Men OA2.06 LB Cryptic Multiple HIV- Infection Revealed by Early, Frequent, and Deep Sampling during Acute Infection Damien C. Tully, Colin B. Ogilvie, Rebecca Batorsky, Karen A. Power, Hunter Bedard, Aaron Seese, Molly Amero, Sue Bazner 2, Jake Tinsley 3, Niall J. Lennon 4, Matthew R. Henn 4, Eric Rosenberg 2, Kenneth H. Mayer 3, Heiko Jessen 5, Marcus Altfeld,6, Todd M. Allen Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 Massachusetts General Hospital, Infectious Disease, Boston, MA, United States, 3 The Fenway Institute, Fenway Health, Boston, MA, United States, 4 The Broad Institute of MIT and Harvard, Cambridge, MA, United States, 5 HIV Clinic Praxis, Jessen, Berlin, Germany, 6 Heinrich- Pette-Institut, Viral Immunology, Hamburg, Germany Background: The global spread of HIV - has been fueled by sexual transmission with the epidemic disproportionately affecting men who sex with men (MSM). As the epidemic in MSM continues unabated, understanding the virus-host interactions responsible for transmission may be critical for the development of an HIV vaccine and other prevention strategies. Methods: To elucidate the nature of the transmitted/founder (TF) virus following rectal transmission, we developed a novel analytical strategy utilizing deep sequencing data from a cohort of 67 acutely infected MSM subjects. Results: Empirical analyses revealed that deep sequencing could not only reliably infer the TF virus but also discriminate between single and multiple HIV infections. Using this approach we found that most transmissions resulted from a single infection with only 6% of individuals exhibiting evidence of multiple variant transmissions. We extended this study to identify signature mutations that may be favored at transmission between viruses originating from heterosexual exposure versus those from MSM. Here, we focused on a comprehensive analysis of Env sequences from 25 early subjects (Fiebig I-III) to discern the genetic imprint on the underlying composition of the viral quasispecies. A number of genetic signatures were identified in gp20 and the gp4 cytoplasmic tail. One signature pattern specifically enriched in TF viruses from MSM was the loss of an N-linked glycosylation site at position 362 in the C3 region adjacent to the CD4 binding site. The loss of this glycosylation motif has previously been associated with chronic infection and implicated in increased cell-to-cell fusion activity and a high apoptosis inducing phenotype. Conclusions: Taken together, these findings provide unique insight into the events of early transmission in MSM and reveal potentially important mechanistic differences that may exist between the different routes of sexual transmission that are not yet fully understood. Gustavo Hernan Kijak,2, Eric Sanders-Buell,2, Agnes-Laurance Chenine,2, Michael Eller,2, Nilu Goonetilleke 3, Rasmi Thomas,2, Sivan Leviyang 4, Elizabeth Harbolick,2, Meera Bose,2, Phuc Pham,2, Celina Oropeza,2, Kultida Poltavee,2, Anne Marie O Sullivan,2, Melanie Merbah,2, Margaret Costanzo,2, Hui Li 5, Will Fischer 6, Feng Gao 7, Leigh Anne Eller,2, Robert J. O Connell 8, Samuel Sinei 9, Lucas Maganga 0, Hannah Kibuuka, Sorachai Nitayaphan 8, Morgane Rolland,2, Bette Korber 6, Francine McCutchan 2, George Shaw 5, Nelson Michael, Merlin Robb,2, Sodsai Tovanabutra,2, Jerome Kim U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Silver Spring, MD, United States, 2 U.S. Military HIV Research Program (MHRP)/ Henry M. Jackson Foundation, Silver Spring, MD, United States, 3 School of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 4 Department of Mathematics and Statistics, Georgetown University, Washington, DC, United States, 5 Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States, 6 Theoretical Biology, Los Alamos National Laboratory, Los Alamos, NM, United States, 7 Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, United States, 8 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 9 Walter Reed Project, Kericho, Kenya, 0 Mbeya Medical Research Programme, Mbeya, Tanzania, United Republic of, Makerere University-Walter Reed Project, Kampala, Uganda, 2 Independent Consultant, Silver Spring, MD, United States Background: In acute HIV- infection (AHI) single genome sequencing (SGS) revealed a strong bottleneck at transmission, with 60-90% of sexual infections being established by a single transmitted/founder (T/F) virus. We combined early and frequent sampling with targeted deep sequencing (TDS) to study viral evolution during AHI. Methods: We studied 7 HIV(-) at entry high-risk RV27 volunteers (2 M and 5 F, all with sexual risk) with documented HIV nucleic acid (NA) conversion after twice-weekly testing. Starting at d2-7 (d0: first NA+ date), we studied 8-9 consecutive plasma samples (mean sampling interval: 4.d; peak viremia: d0-8; -5 samples were from pre-peak viremia) by HIV SGS and TDS (Ion Torrent; limit of detection: 0.5%). Results: 6/7 persons had pre-peak viremia SGS profiles consistent with infection by a single T/F virus. However, in 4 persons, additional variants were detected by TDS: in 3 persons at d2-7 (frequency: %), and in one at d2. Viral populations evolved at dramatic rates, but with different patterns. In #,the minor variant circulated at < 5% until d7, then increased to 57% by d3. In #2, the minor variant increased from 3.5% (d7) to 93% (d2), and then decreased to < 0.5% by d42. In #3 the minor variant was at 0.5-% between d7-6, then became undetectable, but was 53% at d8. In participant #4, 2 minor variants were detected at d2 (0.5-.4%), increasing by d28 to 6-36%, respectively. Full length genetic distances between cognate major and minor variants were.0-2.2%, consistent with acquisition of multiple viruses from the same donor. Inter-variant recombinants were detected from d2 onward. During early AHI, both major and minor variants acquired CTL-escape mutations. Conclusions: We show that in apparent single infections minor variants can occur at levels not detectable by SGS (i.e., cryptic multiple infection). Furthermore these variants contribute to viral evolution, which may have profound implications for HIV pathogenesis, cure, treatment, and vaccine design. ORAL ABSTRACT SESSIONS 99

100 Oral Abstract Sessions Oral Abstract Session 22: Cell and Tissue Models of ARVs for Prevention OA22.0 Oral Maraviroc and Tenofovir for HIV Prevention in Women: An Ex vivo and Translational Approach Melanie R. Nicol, Heather MA Prince 2, Cindi W. Emerson, Julie AE Nelson 2, Kristine B. Patterson 2, Elizabeth J. Geller 2, Myron S. Cohen 2, Angela D.M. Kashuba,2 University of North Carolina at Chapel Hill, Eshelman School of Pharmacy, Chapel Hill, NC, United States, 2 University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, NC, United States OA22.02 Pharmacodynamic Activity in Ectocervical and Colonic Tissue of Dapivirine, Maraviroc, and Combination Topical Gels for HIV Prevention Charlene Dezzutti,2, Sarah Yandura 2, Lin Wang 2, Brid Devlin 3, Jeremy Nuttall 3, Lisa C. Rohan,2 University of Pittsburgh, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 3 IPM, Silver Springs, MD, United States ORAL ABSTRACT SESSIONS Background: Our previous studies demonstrate the protective concentration of tenofovir diphosphate (TFVdp) in vaginal explants is >0-fold higher than in TZM-bl cells. Here we investigate maraviroc s (MVC) efficacy in cells and vaginal explants, and determine the explant s prediction potential of a dose-challenge study from biopsies of volunteers given an oral dose of MVC+ tenofovir disoproxil fumurate (TDF). Methods: TZM-bl cells (n=3) and vaginal explants (n=5 donors) were incubated 24h in MVC ug/mL prior to challenge with HIV- JR- CSF. Combination MVC+tenofovir (TFV) was also used in cells to define the effects of drugs combined. Compared to undosed controls, efficacy was assessed using a luciferase reporter assay in cells, and spliced RNA 24-72h post-inoculation in explants. A dose-challenge study was performed in 6 HIV-, pre-menopausal women administered a single 600mg MVC+600mg TDF dose. 24h post-dose, 4 vaginal+cervical biopsies were collected for viral challenge and evaluated for infection in the same manner as explant tissue. HIV protection was defined as spliced RNA within one standard deviation of background. Results: In vaginal explants, MVC protective efficacy waned after 24h. Within 24h, MVC EC 50 was 9.7ug/mL, which was >000-fold higher than the EC 50 in TZM-bl cells (0.006 ug/ml). Additivity of MVC+TFV was confirmed for HIV protection. The TZM-bl model and the explant model predicted 00% and 6% efficacy, respectively, 24h after a 600mg MVC+TDF dose. In the healthy volunteers, protection was observed in 50% (3/6) of vaginal biposies and 67% (4/6) of cervical biopsies, with 50% (3/6) of women having complete protection against HIV challenge. Conclusions: Similar to TFVdp, cell models overestimated the efficacy of MVC in vaginal explants. Data from TZM-bl cell monolayers over predicted, and tissue explants under predicted, efficacy in a healthy volunteer dose-challenge study. Tissue concentrations at 24h after single high-dose of MVC+TDF were moderately protective against HIV infection. Background: Dapivirine (DPV), a non-nucleoside reverse transcriptase inhibitor, and maraviroc (MVC), a CCR5 antagonist, were formulated into aqueous gels to prevent mucosal HIV transmission. We hypothesize the combination gel will have more potency against HIV infection of mucosal tissue as compared to either single drug gel. Methods: Dilutions of 0.05% DPV, 0.% MVC, and 0.05% DPV/0.% MVC gels were evaluated on polarized ectocervical and colonic mucosal explant cultures exposed to HIV- BaL. After an overnight culture, the explants were washed and medium replenished in the basolateral compartment. Every 3 to 4 days, supernatant was collected and replenished for up to 2 days. HIV- replication was monitored in culture supernatant by p24 ELISA. Results: Dilutions of the gels for ectocervical tissue began at :20 for DPV concentrations of ~75900 nm and MVC concentrations of ~97500 nm, while dilutions for colonic tissue began at :2000; a 00-fold more dilute than what was used for the ectocervical tissue. For ectocervical tissue, 7590 nm of DPV resulted in complete tissue protection while nm of MVC was partially protective (6 of 8 explants showed no HIV replication). The combination gel at 7590 nm of DPV/9750 nm of MVC completely protected the tissue, while 759 nm of DPV/975 nm of MVC was partially protective (6 of 8 explants showed no HIV replication). For colonic explants, DPV gel diluted to 759 nm completely protected the tissue, higher dilutions showed no protection. MVC gel diluted to 975 nm showed no substantial protection of the colonic tissue. The combination gel diluted to 759 nm of DPV/975 nm of MVC completely protected the colonic tissue while the 0-fold higher dilution was partially protective (6 of 8 explants showed no HIV replication). Conclusions: Combining both drugs in a single formulation demonstrated modest synergy. Collectively, these data provide a rationale for further testing of these products as dual compartment microbicides. 00 HIV Research for Prevention 204 HIV R4P

101 Thursday, 30 October Oral Abstract Session 22: Cell and Tissue Models of ARVs for Prevention OA22.03 Expression, Activity, and Regulation of Phosphorylating Enzymes in Genital and Colorectal Tissues and Immune Cells Minlu Hu,2, Tian Zhou,2, Charlene S. Dezzutti,3, Sharon L. Hillier,3, Lisa C. Rohan,2,3 Magee-Womens Research Institute, Pittsburgh, PA, United States, 2 University of Pittsburgh School of Pharmacy, Department of Pharmaceutical Sciences, Pittsburgh, PA, United States, 3 University of Pittsburgh, Department of Obstetrics, Gynecology, and Reproductive Sciences, Pittsburgh, PA, United States OA22.04 Transport and Transport Properties of Tenofovir from Microbicide Gels into Vaginal Tissue: Analysis Using Raman Spectroscopy Oranat Chuchuen, Marcus H. Henderson, Marinella G. Sandros 2, Angela D.M. Kashuba 3,4, David F. Katz,5 Duke University, Biomedical Engineering, Durham, NC, United States, 2 University of North Carolina at Greensboro, Nanoscience, Greensboro, NC, United States, 3 University of North Carolina at Chapel Hill, Eshelman School of Pharmacy, Chapel Hill, NC, United States, 4 University of North Carolina School of Medicine, Department of Infectious Diseases, Chapel Hill, NC, United States, 5 Duke University, Department of Obstetrics and Gynecology, Durham, NC, United States Background: Studies of oral tenofovir (TFV) have revealed that TFV and tenofovir diphosphate (TFV-DP) levels are 00 higher in colorectal tissue than in cervical/vaginal tissue after a single oral dose. Multiple phosphorylating enzymes (PEs) play a role in TFV activation. However, limited data is available regarding the expression and activity of these enzymes in the female genital and colorectal tissue relative to immune cells. Methods: mrna expression for 7 PEs (AK2, AK4, NME, NME2, CKMT, CKMT2, CKB) in fresh surgical human tissue samples (cervical n=6, vaginal n=5, colorectal n=5), a vaginal epithelial cell line (VK2), and a T cell line (PM), was evaluated using qrt-pcr. Intracellular TFV-DP formation was tested in VK2 and PM cells with or without medroxyprogesterone acetate (MPA) and progesterone (P4) using an LC- MS/MS method. Differences in TFV-DP conversion were assessed using Student s t-test. Results: Vaginal, ectocervical, and colorectal tissues had similar expression of PEs except for AK2, which was present at 5-28 higher levels in colorectal tissue than in ectocervical or vaginal tissues (p< 0.05). The vaginal epithelial cell line was shown to have 0-0,000 higher expression of CKB, CKMT, CKMT2, AK2, and AK4 as compared to levels found for the T cell line (p< 0.05). MPA treatment resulted in a 3-fold increase in TFV-DP in the epithelial cell line (p< 0.0) and a 30% decrease in the T cell line (p< 0.0) as compared to controls. P4 treatment resulted in a nearly 4-fold increased TFV-DP level in the epithelial cell line (p< 0.0) and no change in the T cell line. Conclusions: The increased levels of AK2 in colorectal tissue suggest that AK2 may contribute to the increased levels of TFV-DP observed in colorectal tissues. The higher level of PEs observed in vaginal epithelial cell line compared to T cell line, suggests that TFV-DP found in tissues may be predominantly associated with epithelial cells. The impact of reproductive hormones on PEs warrants further investigation. Background: Common drug release assays use a liquid sink receptor compartment. Permeability assays measure net transport through tissue specimens of varying thickness. These do not give concentration vs. depth in tissue, nor distinguish drug partitioning at the vehicle-tissue interface from rate of transport in tissue. We developed a rapid, noncontact method using confocal Raman spectroscopy to measure drug partitioning and concentration vs. depth in intact tissue layers (epithelium vs. stroma) and to translate such data to drug diffusion coefficients. We report here on results for Tenofovir released from its clinical gel. Methods: Fresh porcine vaginal tissue specimens were treated with % Tenofovir gel in a Transwell assay for 2-8 hr at 37 C. Gel was applied to either epithelial or stromal tissue surfaces. Results for spatiotemporal concentration profiles were fit to a drug diffusion model to obtain diffusion coefficients in epithelium and stroma. To determine partition coefficients, tissue specimens were incubated by submersion in % Tenofovir gel and equilibration over 6 h. Results: Tenofovir concentrations exhibited diffusion-like time- and depth-dependent distributions in tissue. Diffusion and partition coefficients in epithelium ranged 7x x0-8 cm 2 /s, and , respectively. Initial measurements gave log increase in diffusion coefficient in stroma. Measurements were referenced to classical permeability data. Conclusions: This standardizable label-free method characterizes drug concentration distributions in tissue and gel vehicles, determining the fundamental gel-tissue partition coefficient and diffusion coefficients in gel, epithelium and stroma. Results suggest that the epithelium presents a potential rate-limiting barrier to Tenofovir permeation across vaginal mucosa. This is more incisive and pharmacologically useful information than results of traditional methods that do not distinguish transport across the two layers; transport parameters can be input to computational PK models. ORAL ABSTRACT SESSIONS 0

102 Oral Abstract Sessions Oral Abstract Session 22: Cell and Tissue Models of ARVs for Prevention OA22.05 OA22.06 LB ORAL ABSTRACT SESSIONS Mucosal Tissue Explants as Surrogates for in vivo Efficacy of Microbicides Carolina Herrera, Ronald Veazey 2, Angela Kashuba 3, Javier García Pérez 4, José Alcamí 4, Karl Malcolm 5, Robin Shattock Imperial College, Infectious Diseases, London, United Kingdom, 2 Tulane National Primate Research Center, Tulane, LA, United States, 3 The University of North Carolina, Chapel Hill, NC, United States, 4 Instituto de Salud Carlos III, Madrid, Spain, 5 Queen s University Belfast, Belfast, United Kingdom Background: Validity of the non-human primate (NHP) model is often questioned due to the lack of correlation with clinical trials in humans. We hypothesize that in vivo dosing of candidate microbicide conferring protection against HIV-/SIV transmission in mucosal sites, can be predicted with a surrogate model of ex vivo tissue explant cultures, through intra-tissular drug pharmacological measurements and ex vivo infection of tissue explants Methods: Gel-formulated nucleotide reverse transcriptase inhibitor tenofovir (TFV), and entry inhibitor, maraviroc (MVC), alone or in combination at fully and partially inhibitory doses were tested. Rhesus macaque and human cervicovaginal and colorectal tissue explants were exposed to gels for h followed by addition of virus for 2 h. Wild type isolates (BaL, YU.2, SIVmac32H and RT-SHIV) and NRTI-escape mutants (point mutations K65R +/- M84V in YU.2 and SIVmac32H) were used. Tissue concentrations (PK/PD parameters) of TFV, TFV diphosphorylated (dp) and MVC were assessed at different time points during 5 days of culture. Infection was assessed by measurement of p24/p27 in culture supernatants Results: In NHP and human explants high tissue drug levels and low rates of elimination were predictive of drug antiviral efficacy with negative linear dose-response relationships observed between explant drug levels and p24/p27 concentrations. Greater potency with combination gels was seen in NHP than in human tissue. Opposite drug distribution was observed between both species with higher PK values in colorectal than in cervicovaginal explants in NHP. Greater loss of viral replication fitness was seen with SIV RT mutations in NHPs than in human explants with mutant HIV- Conclusions: Ex vivo dose-challenge studies with human and NHP explants confirmed robustness of the explant model and its potential as surrogate for in vivo studies refining the prediction of candidate microbicides efficacy in clinical trials and reducing the number of NHP euthanized Predicting Effective Truvada PrEP Dosing Strategies With a Novel PK-PD Model Incorporating Tissue Active Metabolites and Endogenous Nucleotides (EN) Mackenzie L. Cottrell, Kuo H. Yang, Heather M.A. Prince, Craig Sykes, Nicole White, Stephanie Malone, Evan S. Dellon 2, Ryan D. Madanick 2, Nicholas J. Shaheen 2, Julie A. Nelson 3, Ronald Swanstrom 3, Kristine B. Patterson 2, Angela D.M. Kashuba University of North Carolina Eshelman School of Pharmacy, Chapel Hill, NC, United States, 2 University of North Carolina School of Medicine, Chapel Hill, NC, United States, 3 University of North Carolina Center for AIDS Research, Chapel Hill, NC, United States Background: Failure of daily tenofovir(tfv) disoproxil fumarate(tdf)±emtricitabine(ftc) chemoprophylaxis in women has been attributed to poor adherence. Yet as few as 2 doses/week has been shown effective in MSM. Here, we provide dosing strategy predictions for female genital tract and colorectal tissue(rt) utilizing a PK-PD model that incorporates mucosal tissue TFV diphosphate(dp) and FTC triphosphate(tp) PK, concentrations of ENs(dATP and dctp) and their respective molar ratios that prevent HIV infection. Methods: TZM-bl cells were treated with µg/ml TFV and µM FTC for 24h before HIV- JR-CSF challenge. TFVdp, FTCtp, datp and dctp were quantified and infection measured by luciferase. R was used to fit an Emax model of TFVdp:dATP or FTCtp:dCTP vs %inhibition of infection. 48 women were given a single dose of TDF or FTC at 50, 00 or 200% of the licensed dose. TFVdp, FTCtp, datp and dctp were measured in cervical(ct), vaginal(vt) and RT tissue over 48h using LC-MS/MS. NONMEM7.3 was used to fit a population PK model and subsequent monte-carlo simulations. Results: From TZM-bl cells, the EC 90 (0.086 for TFV and for FTC;p< 0.00) was used as the clinical target. An 8 compartment linear model best described tissue PK. By 7 days FTC200mg daily achieved ratios >EC 90 for >85, 50 and 75% of the population in VT, CT and RT; and TDF300mg daily achieved ratios >EC 90 for < 50%, < 50% and 00% of the population in VT, CT and RT. For RT 2 doses/week maintained >EC 90 in 00% of the population. TDF+FTC maintains >EC 90 for >75 and 50% of the population in VT and CT with daily dosing and 00% in RT with 2 doses/week. Conclusions: This study is the first to model TDF/FTC dosing strategies utilizing only in vitro and mucosal tissue pharmacokinetic data. This model predicts available clinical trial data, whereby TDF/FTC is ~70% effective in women with 80% adherence and >90% effective in MSM with ~30% adherence. We believe a-priori utilization of this novel paradigm can enhance clinical trial design and outcomes. 02 HIV Research for Prevention 204 HIV R4P

103 Thursday, 30 October Oral Abstract Session 23: Pregnancy Intentions, Safe Conception and PMTCT OA23.SY PrEP for Women: Indications and Worldwide Implementation for Women OA23.0 Evaluation of HIV- Neutralizing Antibodies in Maternal-infant Transmission in Thailand Erika Aaron Drexler University College of Medicine, United States PrEP has been shown to be efficacious in populations of women and offers a promising female-controlled method of prevention. Women continue to be a risk of HIV acquisition due to biological, behavioral, and cultural factors with unacceptable rates of new infection. Currently recommended sexual HIV-prevention strategies for women include abstinence, condom use, and treatment as prevention; all require the willingness of the male partners and none allows for conception. HIV-affected couples who want to have children face the challenge of preventing HIV transmission to the sero-negative partner within a serodiscordant couple. While there are reproductive technologies that can help HIV-affected couples to safely conceive with minimal risk of HIV transmission to their partner, for most couples such technologies are neither geographically nor economically accessible. Periconception PrEP may be a useful adjunct for serodiscordant couples. This presentation will describe the benefits and potential role of PrEP for women, the use of periconception PrEP for HIV-serodiscordant couples, and the need to improve and scale up the implementation of PrEP in the US and worldwide. Lindsay Wieczorek,2, Brittani Barrows,2, Agnès-Laurence Chenine,2, Martine Braibant 3, Kriengkrai Srithanaviboonchai 4, Panita Pathipvanich 5, Shelly Krebs,2, Nelson L. Michael,6, Sodsai Tovanabutra,2, Jerome H. Kim,6, Merlin Robb,2, Victoria Polonis,6 Military HIV Research Program, Bethesda, MD, United States, 2 Henry M Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 3 Université François-Rabelais, Tours, France, 4 Chiang Mai University, Chiang Mai, Thailand, 5 Lampang Regional Hospital, Lampang, Thailand, 6 Walter Reed Army Institute of Research, Silver Spring, MD, United States Background: The role of neutralizing antibody (NAb) in mother-to-child transmission (MTCT) of HIV- remains unclear. Previous studies suggest that maternal NAb might reduce HIV- transmission. Higher NAb titers to MBA, a CRF0_AE strain with an unusually long V2 domain, were found to correlate with lower rates of intrapartum MTCT. However, findings from different MTCT studies are inconsistent, and further work is required to clarify the impact of NAb in MTCT. Methods: In this study, we evaluated NAb breadth and potency in plasma from 0 HIV+, ART-naïve mothers (22 transmitters and 79 nontransmitters) collected at delivery, and from 5 of their infants (6 HIV+ and 35 HIV-) collected two months after birth. Pseudovirus (PV) assays were employed using a panel of six CRF0_AE isolates, including MBA and RV44 vaccine strains TH023 and CM244. NAb activity is reported as ID50 titer or positive area under the curve (+AUC), useful for evaluating samples with low NAb activity. Results: Contrary to previously published results, maternal geometric mean NAb titers and +AUC trended higher for transmitters compared to non-transmitters for five of the six PV tested (including MBA), with a significant difference observed for CM244 (p=0.047). Maternal NAb breadth was also increased in transmitters (p=0.047) and directly correlated with viral load (p=0.037). As expected, infant NAb +AUC was increased for HIV+ infants compared to those that did not seroconvert for two pseudoviruses CM244 (p=0.042) and (p=0.09). The relationship between mother and infant NAb activity is currently being evaluated. Conclusions: Greater magnitude maternal NAb titers were unexpectedly associated with MTCT transmission of HIV, but correlated with higher viral load. Further work is required to understand the development, specificity, and function of NAb in MTCT of HIV. ORAL ABSTRACT SESSIONS 03

104 Oral Abstract Sessions Oral Abstract Session 23: Pregnancy Intentions, Safe Conception and PMTCT OA23.02 PMTCT Adherence in Pregnant South African Women: The Role of Depression, Social Support, Stigma and Structural Barriers to Care OA23.03 Barriers and Promoters to Uptake of Safer Conception Strategies among HIVserodiscordant Couples with Fertility Intention in Mbarara, Uganda ORAL ABSTRACT SESSIONS Christina Psaros, Nzwakie Mosery 2, Jennifer A. Smit 2, Faith Luthuli 3, Janna R. Gordon 4, Ross Greener 3, Kara Bennett 5, David R. Bangsberg 6, Steven A. Safren Massachusetts General Hospital / Harvard Medical School, Psychiatry, Boston, MA, United States, 2 MatCH Research (Maternal, Adolescent and Child Health Research), Obstetrics and Gynecology, Durban, South Africa, 3 MatCH Research (Maternal, Adolescent and Child Health Research), Durban, South Africa, 4 Massachusetts General Hospital, Psychiatry, Boston, MA, United States, 5 Bennett Statistical Consulting, Inc., Ballston Lake, NY, United States, 6 Massachusetts General Hospital / Harvard Medical School, Center for Global Health, Boston, MA, United States Background: Depression is a robust predictor of non-adherence to antiretroviral therapy, essential in PMTCT. Women in resource-limited settings are likely to face additional barriers to PMTCT adherence, including stigma and structural barriers. While structural barriers may be circumvented by social support; depression and stigma may make access difficult. Understanding modifiable factors that contribute to PMTCT adherence can inform interventions. Methods: 67 HIV-infected women enrolled in PMTCT (median age 28 years) completed an interview at > 28 weeks of pregnancy assessing depression, stigma, social support and structural barriers to PMTCT. An adherence score was created using principal components analysis on the response to four questions assessing adherence over the past 30 days. Depression was defined as a Hopkins score >.75 and was examined as a predictor of the adherence score in a linear regression model. Separate linear regression models also examined relationships between () social support and structural barriers (income and time spent traveling to clinic) and (2) depression and stigma as predictors of social support. Results: Participants with elevated depressive symptoms had significantly lower adherence scores (p< 0.0). Neither income (p=0.0) nor time spent traveling to clinic (p=0.28) predicted adherence; thus, moderation with social support was not examined. Depression significantly predicted social support (est= p< 0.0): those with elevated depressive symptoms had a lower social support score. Similarly, a higher stigma score was significantly associated with a lower social support score (est=-0.09, p< 0.0). Conclusions: While PMTCT programs are effective, adherence to these services is suboptimal. Depression may play an important role in adherence to these behaviors. HIV infected pregnant women with elevated depressive symptoms may also suffer from low social support and high stigma; interventions targeting these factors may support maternal and fetal health. Angela Kaida, Jasmine Kastner, Courtney Ng 2, Naomi Sanyu 3, Adrine Kusasira 3, Jerome Kabakyenga 3, David R. Bangsberg 3,4, Lynn T. Matthews 4 Simon Fraser University, Faculty of Health Sciences, Vancouver, BC, Canada, 2 Massachusetts General Hospital (MGH), Center for Global Health, Boston, MA, United States, 3 Mbarara University of Science and Technology, Mbarara, Uganda, 4 MGH Center for Global Health & Division of Infectious Disease, Boston, MA, United States Background: We investigated barriers and promoters to uptake of a safer conception approach to pregnancy among HIV sero-discordant couples in Mbarara, southwestern Uganda. Methods: We recruited HIV-infected men and women (index) receiving antiretroviral therapy (ART) from the Uganda Antiretroviral Rural Treatment Outcomes cohort who reported an uninfected or unknown status partner (partner), serostatus disclosure to the partner, and personal or partner desire for a child within 2 years. We conducted 40 separate in-depth interviews with 20 couples to explore periconception risks and awareness of specific safer conception strategies. Data were translated, transcribed, and analyzed using content analysis. Results: 2/20 index participants were women, with median age of 36 yrs [IQR 29-4], and median recent CD4 of 433 cells/mm 3 [IQR: ]. Median partner age was 34yrs [IQR 30-40]. Awareness of HIV prevention strategies beyond condoms and abstinence was limited, however, some participants described timed intercourse and ART as prevention as ways to reduce HIV transmission. Participants were motivated to learn more about safer conception strategies. Key barriers included limited couple communication about childbearing plans and understanding of HIV sero-discordance. Fatalism about eventual HIV acquisition by the uninfected partner or a sense of protection due to strong blood or God s will were common perceptions that decreased motivation to practice HIV prevention. Many participants prioritized pregnancy with minimal perceived options for reducing HIV risk. The more vulnerable partner (HIV-infected and/or female) was often eager to pursue pregnancy to secure the relationship, regardless of HIV acquisition or transmission risks. Conclusions: Awareness of ART for prevention and high interest in other safer conception strategies presents opportunity to encourage mutual status disclosure, contravene normative expectations of eventual seroconversion, and promote strategies to minimize periconception HIV risks. 04 HIV Research for Prevention 204 HIV R4P

105 Thursday, 30 October Oral Abstract Session 23: Pregnancy Intentions, Safe Conception and PMTCT OA23.04 I Would Say it Does Concern Me and on the Other Hand it Doesn t. Perceptions of South African Learners Experiences with Sex, Pregnancy, and HIV Cecilia Milford, Lizzie Moore, Mags Beksinska, Muriel Kubeka, Kedibone Sithole, Sibusiso Sibiya, Faith Smangele Luhthuli, Jennifer Smit MatCH Research, Department of Obstetrics and Gynaecology, University of the Witwatersrand, Durban, South Africa Background: HIV/AIDS, sexually transmitted infections (STIs) and teenage pregnancy are concerns for South Africa s youth. Adolescent pregnancy is a major cause of interrupted schooling and drop-out despite pregnant learners being protected by law. Incomplete education and early pregnancy are risk factors for HIV acquisition. This study reports on perceptions of learners experiences with sex, pregnancy, and HIV. Methods: Focus groups were held with male and female learners (n=4, 4 groups), parents (n=9, 2 groups), educators (n=, 2 groups) and community members (n=9, 2 groups) recruited through two schools in ethekwini District, KwaZulu-Natal, South Africa. Discussions were transcribed, translated and data coded. Results were organised according to key themes and NVivo used to facilitate data analysis. Results: Almost half the learners (n=7), aged 6-2, had initiated sex, most common age of first sex was 5 (n=5). Four learners had been pregnant. Substance use, transactional sex and low/inconsistent condom use were the main risk factors for pregnancy and STIs. Although learners knew about HIV, some were not concerned about it, there is something you can use to reduce it, however stigma was a barrier to accessing HIV-related services. While teachers discussed HIV with learners, across groups, most felt that parents should provide advice on abstinence, protection during sex and monogamy. However some parents lacked information and others feared discussing HIV with their children. Teenage pregnancy was reportedly common in schools, mostly unplanned but some perceived to access government grants. Pregnancy led to drop-out and gaps in schooling. Conclusions: Teenage learners are practicing unprotected sex despite being educated about HIV and pregnancy. Barriers to accessing services put them at risk. There is a need for improved access to services, better access to information for parents, and improved relationships with parents to address gaps and influence behaviour. ORAL ABSTRACT SESSIONS 05

106 Oral Abstract Sessions Oral Abstract Session 24: Mucosal Responses OA24.0 Vaccine Induced Responses in a SIV Model Can Impact Challenge Outcomes OA24.02 Local HIV-specific IgA Antibody Production in the Penile Urethra Mucosal Compartment ORAL ABSTRACT SESSIONS Megan Wise, Michele Kutzler, Natalie Hutnick, Zina Moldoveanu 2, Meredith Hunter 3, Jian Yan 4, Bapi Pahar 3, Devin Myles, Amir Khan 4, David Montefiori 5, Michael Betts, Niranjan Sardesai 4, Jiri Mestecky 2, Preston Marx 3, David Weiner University of Pennsylvania, Philadelphia, PA, United States, 2 University of Alabama at Birmingham, Birmingham, AL, United States, 3 Tulane National Primate Research Center, Covington, LA, United States, 4 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States, 5 Duke University, Durham, NC, United States Background: The use of cytokine gene adjuvants to tailor the immune response is a strength of EP DNA vaccination. This has been established through the recent HVTN080 trial which demonstrated the potency of IL-2 with EP delivered DNA to drive T cell responses. However, for an HIV vaccine it is important to also induce strong humoral responses. To address the possible role of serum and vaginal IgA in HIV acquisition we utilized an adjuvanted DNA vaccine previously shown to drive IgA induction in a non-human primate vaginal challenge model. Methods: Groups of 5 Indian rhesus macaques received a psivmac239 gag/pol and psive660 gp20 alone, or with plasmids - pccl25 (TECK), pccl27 (CTACK) or pccl28 (MEC), genetic adjuvants, at weeks 0, 6, 2, 8 and 48. Animals were challenged with 500 TCID SIVsmE660 intravaginaly twice a week for two weeks for 4 challenges. Results: We observed higher vaginal IgA titers in gene adjuvanted animals compared with DNA vaccine alone. Following challenge, we observed an overall protection rate of 68% for all vaccinated animals. However, this protection rate was different for each vaccination regiment. Animals vaccinated with CCR0Ls, (CCL27 and CCL28,) exhibited robust control of set point viremia and chronic viremia (p< 0.05) with 89% of animals controlling infection compared with only 40% in unadjuvanted animals and 4% in naïve challenge controls. However, CCR9L (CCL25) vaccinated animals resisted challenge in 60% of animals. Irrespective of vaccine group, animals that controlled viremia had the highest vaginal IgA and IgG levels post-vaccination. Conclusions: Inclusion of immune plasmid adjuvants encoding mucosal chemokines in EP DNA vaccine regiments can improve challenge outcomes. Collectively these adjuvant approaches likely have importance for the development of next generation DNA vaccines and the data illuminates the need for continued research into the role of vaginal antibodies and protection from viral infection in NHP models. Kadryn Kadasia, Joseph Politch, Matt Schoen 2, Amy Chung 2, Galit Alter 2, Deborah Anderson Boston University, School of Medicine, Boston, MA, United States, 2 Ragon Institute of MIT, MGH and Harvard, Cambridge, MA, United States Background: Whereas the genital mucosa serves as the first immune barrier to sexually transmitted pathogens, little is known about the production and role of antibodies in the human mucosal environment. Previous studies have shown an abundance of IgG + and IgA + plasma cells associated with penile urethral glands suggesting that this is a site of local immunoglobulin (Ig) production in men. We compared HIV-specific Ig isotypes (A, G) and subclasses (G, G3) in blood plasma (BP), seminal plasma (SP) and urethral preejaculatory (PE) secretions from HIV-infected men and controls to determine whether a distinct population of HIVspecific antibodies is present in the genital mucosa during HIV infection. Methods: A multiplex bead-based Luminex assay incorporating a panel of seven HIV-specific antigens was used to compare titres, isotypes, and subclasses of HIV-specific antibodies in PE, SP and BP from nine HIVinfected men and nine healthy controls. We also conducted antibodydependent cell phagocytosis (ADCP) and cytotoxicity (ADCC) assays to evaluate antibody function in the different compartments. Results: There was an abundance of HIV-specific IgG in all three fluids, with similar titres, subclasses and specificity profiles. Similarly, ADCC and ADCP activity did not differ between the three sample types. In contrast, a subset of PE samples from HIV-infected men had significantly higher (5-50X) anti-gp4 IgA titres than found in SP and BP, providing evidence for local urethral production of HIV-specific IgA during HIV infection. Conclusions: IgG HIV antibody profiles in male genital secretions reflect those in the systemic circulation, whereas IgA HIV antibodies may be locally produced in the urethral mucosa. More research is needed on strategies to elicit urethral antibody production with vaccines, and the function of IgA antibodies in HIV prevention. 06 HIV Research for Prevention 204 HIV R4P

107 Oral Abstract Session 24: Mucosal Responses OA24.03 Targeting Mucosal Fc-Fc Receptor Interactions as Vaccine Strategy against Mucosal HIVtransmission Thursday, 30 October OA24.04 DNA and Protein Co-immunization Improves the Magnitude, Longevity, and mucosal Dissemination of Immune Responses Magdalena Sips,2, Marina Krykbaeva, Brittany Bowman, Thomas Diefenbach, Douglas Kwon, Peter Brouckaert 2, Galit Alter Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 Ghent University, Ghent, Belgium Background: To ultimately end the AIDS pandemic, effective means of preventing transmission by its principal genital and rectal routes must be developed. Non-neutralizing antibody (nnab) responses, which may be easier to induce than broadly neutralizing antibodies via vaccination, have shown some protection, pointing toward a role for nnab functions, including Fc-Fc receptors (FcRs) interactions. The protective efficacy of Fc-effector function is critically dependent on innate immune cells; therefore we sought to define cell repertoires expressing FcRs in the female genital tract (FGT) and intestinal tract (IT). Methods: Fixed tissue sections (rectum, vagina, cervix, uterus, lymph node) were stained for natural killer (NK) cells, macrophages, neutrophils and Fc receptors, FcγR: I, II, III and FcαR. HIV+ biopsy samples from IT and lymph nodes were examined for changes in FcR repertoires. Flow cytometric evaluation of FcR+ cells was performed on freshly isolated cells from enzymatically digested colon and cervical tissues. Results: NK cells were very infrequent in IT and FGT. Moreover, none of the FcR was found on NK cells, suggesting that mucosal NK cells have limited capacity to mediate antibody-driven-effector functions. In contrast, macrophages and neutrophils were present at much higher frequencies in IT and FGT and were detected close to epithelial layers. They robustly expressed FcRs with macrophages expressing FcγRI, FcγRII, FcαR but not FcγRIII and neutrophils expressing FcγRIII and FcαR. Changes in expression pattern in HIV-infected subjects suggest specific antibody therapeutic opportunities for harnessing Fc-FcR interactions. Conclusions: The ability of nnabs to provide protection at mucosal barriers is centrally linked to FcR+ cells available within vulnerable tissues. Surprisingly, our data suggest that non-adcc mediated mechanisms, such as phagocytosis and neutrophil activation, are more abundant and potentially represent important mechanisms by which vaccine induced nnabs may offer protection. George N. Pavlakis, Jinyao Li, Antonio Valentin, Rashmi Jalah, Vainav Patel, Margherita Rosati, Viraj Kulkarni, Candido Alicea, Diego A. Vargas-Inchaustegui 2, Yongjun Guan 3, David Venzon 4, Niranjan Sardesai 5, Timothy R. Fouts 6, Abraham Pinter 7, Marjorie Robert-Guroff 2, David C. Montefiori 8, Xiaoying Shen 8, Georgia D. Tomaras 8, Barbara K. Felber National Cancer Institute at Frederick, Frederick, MD, United States, 2 National Cancer Institute, NIH, Bethesda, MD, United States, 3 Institute of Human Virology, Baltimore, MD, United States, 4 National Cancer Institute, NIH, Biostatics and Data Management Section, Bethesda, MD, United States, 5 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States, 6 Profectus Biosciences, Inc., Baltimore, MD, United States, 7 Public Health Research Institute, University of Medicine and Dentistry of New Jersey, Newark, NJ, United States, 8 Duke University Medical Center, Durham, MD, United States Background: Determining the quality and longevity of vaccine-induced immune responses is essential for improving the prospects of AIDS vaccines. DNA and protein (inactivated viral particles) co-immunization regimen induced systemic and mucosal Ab responses, which correlated with slower virus acquisition upon challenge, and potent T cell responses providing protection against chronic viremia. We are evaluating different regimens of DNA&protein vaccines using purified SIV or HIV- Env to further dissect humoral and cellular responses including magnitude, breadth and mucosal dissemination. Methods: Macaques were vaccinated using DNA&protein coimmunization regimen in the presence of IL-2 DNA, coinjected into the same muscle. Humoral and cellular responses were monitored in blood and different tissues. Results: Co-immunization strategy of DNA&protein induced rapid and high humoral responses while maintaining robust cellular responses typically obtained with DNA vaccines. The vaccine induced Ab against both homologous and heterologous Env; high binding titers against scaffolded V/V2 env region; efficient dissemination to mucosal sites; high Env-specific IgG in saliva and Env-specific IgG and IgA in rectal mucosa. Analysis of cellular responses revealed the presence of cytotoxic memory T cells against several viral proteins. These cellular responses disseminated systemically as demonstrated by their presence not only in blood and lymphoid tissues, but also in bone marrow, liver, lung (effector site) and, importantly, rectal and vaginal mucosa. The longevity of the cellular responses induced by this co-immunization regimen was significantly improved, with SIV-specific T cells detected >5 yrs after the vaccination. Conclusions: Intramuscular DNA&protein co-delivery increases the magnitude and longevity of systemic and mucosal humoral immune responses in immunized macaques and is proposed as a practical and efficient method for human vaccination. ORAL ABSTRACT SESSIONS 07

108 Oral Abstract Sessions Oral Abstract Session 24: Mucosal Responses OA24.05 Antibody Isotypes Differ in their Capacity to Bind, Capture and Aggregate HIV- Virions OA24.06 LB Transcriptional Signatures of Viral Control in HIV- Infected South African Women ORAL ABSTRACT SESSIONS Sandra G. Okala, Deborah F. King, Paul M. Rogers, Robin J. Shattock Imperial College London, Medicine, London, United Kingdom Background: Recently, Env-specific monomeric (m)iga was correlated with increased risk of HIV- infection in the RV44 vaccine trial and inhibited IgG effector functions. In contrast, mucosal dimeric (d)iga has been associated with resistance to infection in highly exposed uninfected individuals, and viral aggregation by Env specific antibodies in mucosal secretions has been often proposed as an alternative mechanism to block HIV- infection at the portal of viral entry. The current study was designed to determine whether anti-hiv- IgG, miga2 and diga2 with the same epitope specificity differ in their ability to bind, capture and aggregate HIV- BaL. Methods: Bio-Layer interferometry was used to measure kinetics parameters of the different forms of b2, CH3, 2F5 and 7B2 mabs to soluble HIV- BaL gp40 Env. Virus capture by the panel of mabs was quantified by ELISA and antibody mediated viral aggregation (AMVA) was determined using Nanoparticle Tracking Analysis. Results: Interestingly, IgGs captured more virions than both migas and digas with the same epitope specificity. Although capture did not correlate with binding affinity (Kd), data indicates that virions capture correlated with association rate constant (kon). No relationships was found between binding affinity and AMVA, however a significant correlation was observed between the number of binding sites and the proportion of aggregates, highlighting improved aggregation capacity of dimeric mabs (P= 0.008). Further, the data demonstrated that AMVA was dependent on epitope accessibility with a classical prozone effect. Conclusions: Overall, our findings indicate that antibody isotype influences effector functions, with greater capacity of IgGs to capture HIV- particles and Env specific digas to induce viral aggregation. Thus, the ability of an antibody-based vaccine to prevent HIV- infection may be dependent on the isotype of the antibody, and the effector functions most relevant to the biological compartment. Nikki L. Gentle,2, Sarah Djebali 3, Neil A. Martinson 4, David Spencer 5, Roderic Guigo 3,6, Caroline T. Tiemessen,2 University of the Witwatersrand, Faculty of Health Sciences, Johannesburg, South Africa, 2 National Institute for Communicable Diseases, Centre for HIV and STIs, Johannesburg, South Africa, 3 Centre for Genomic Regulation, Bioinformatics and Genomics Group, Barcelona, Spain, 4 University of the Witwatersrand, Perinatal HIV Research Unit, Johannesburg, South Africa, 5 Right to Care, Johannesburg, South Africa, 6 Universitat Pompeu Fabra, Barcelona, Spain Background: The characterization of host genetic factors that allow small subsets of individuals to naturally suppress HIV- viral replication in the absence of antiretroviral treatment remains a priority, as an understanding of the immune mechanisms employed by these individuals to control viral replication could provide insights into potential therapeutic targets. Methods: Therefore, in order to identify genes involved in the control of HIV- viral replication during chronic infection, mrna was extracted from peripheral blood mononuclear cells isolated from 4 Black female South African HIV- controllers. Paired-end RNA sequencing of 00 bp fragments was performed using the HiSeq 2000 and the data obtained were processed using the GRAPE analysis pipeline. Differences in gene expression were evaluated using the R/Bioconductor package, DESeq; with genes exhibiting at least a 2-fold difference in expression and p-values of p< 0.00 considered to be differentially expressed. Results: Genes found to be differentially expressed between the eight individuals with viral loads below 400 copies/ml and six individuals with viral loads above 400 copies/ml included several previously implicated in the control of chronic viral infections. These include LAG3, a regulator of T cell responsiveness, and genes involved in the regulation of the interferon type I antiviral response (for example IFI6, IFIT3, IFI44 and OASL). Several of the identified genes also encoded as yet functionally uncharacterized large intergenic non-coding RNAs, whose impact on HIV- viral control remains to be evaluated. Conclusions: Collectively, these data describe a transcriptional signature of immune activation in chronic HIV- infection, which suggests the involvement of innate immune mechanisms previously shown to display substantial sex-based differences. However, whether these mechanisms directly regulate HIV- viral replication, or rather are activated in response to increased viral loads, remains to be established. 08 HIV Research for Prevention 204 HIV R4P

109 Oral Abstract Session 25: Adjuvants and Immunogens Thursday, 30 October OA25.0 Adjuvant Dependent Mucosal V2 Responses and RAS Activation in Vaccine Induced Protection from SIV mac25 Acquisition RAS, a signal transducer that facilitates cross talk among B-cells, T-cells and antigen presenting cells, was demonstrated to be a biomarker of vaccine efficacy in the alum group. Conclusions: These data highlight the importance of the quality of the mucosal antibodies to V2 in protection and suggest that activation of RAS may constitute a novel approach to improve vaccine efficacy against HIV. Monica Vaccari, Shari N. Gordon, Slim Fourati 2, Luca Schifanella,3, Mark Cameron 2, Brandon F. Keele 4, Xiaoying Shen 5, Georgia Tomaras 5, Erik Billings 6, Mangala Rao 6, Namal P.M. Liyanage, Diego A. Vargas-Inchaustegui 7, Steve Whitney 8, Melvin N. Doster, Nicolo Binello, Poonam Pegu,6, David C. Montefiori 9, Kathryn Foulds 0, David S. Quinn 0, Mitzi Donaldson 0, Frank Liang 0, Karin Lore 0, Mario Roederer 0, Richard Koup 0, Adrian McDermott 0, Zhong-Min Ma, Miller Christopher, Tran B. Phan 2, Donald N. Forthal 2, Matthew Blackburn, Francesca Caccuri, Guido Ferrari 9, Marjorie Robert- Guroff 7, Silvia Ratto-Kim 6, Jerome Kim 6, Nelson Michael 6, Sanjay Phogat 3, Susan W. Barnett 4, James Tartaglia 3, David Venzon 5, Donald M. Stablein 6, Rafick-Pierre Sekaly 2, Genoveffa Franchini National Cancer Institute, Bethesda, MD, United States, 2 Vaccine & Gene Therapy Institute, Port St. Lucie, FL, United States, 3 Universita degli Studi di Milano, Milan, Italy, 4 National Cancer Institute SAIC, AIDS and Cancer Virus Program, Frederick, MD, United States, 5 Duke Human Vaccine Institute, Durham, NC, United States, 6 Walter Reed Army Institution, U.S. Military HIV Research Program, Silver Spring, MD, United States, 7 National Cancer Institute, Immune Biology of Retroviral Infection Section, Bethesda, MD, United States, 8 ABL, Rockville, MD, United States, 9 Duke University, Durham, NC, United States, 0 Vaccine Research Center, Bethesda, MD, United States, California National Primate Research Center, Davis, CA, United States, 2 University of California, Irvine School of Medicine, Irvine, CA, United States, 3 Sanofi Pasteur, Swiftwater, IL, United States, 4 Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United States, 5 National Cancer Institute, Biostatistics and Data Management Section, Bethesda, MD, United States, 6 The EMMS Corporation, Rockville, MD, United States Background: The RV44 HIV vaccine trial resulted in limited, but significant protection, from HIV acquisition. Serum antibodies directed to the Env variable regions and 2 (V/V2) inversely correlated with the risk of HIV- infection and sieve analysis demonstrated immunologic pressure on two regions of the V2. Prior macaque studies demonstrated that a similar vaccine for SIV (ALVAC-SIV) induced protection from SIV mac25 acquisition in a low dose neonatal challenge model, but not in adult high dose challenge models. Methods: We challenged ALVAC-SIV/gp20/alum vaccinated adult macaques intrarectally with repeated low doses of SIV mac25 in a study powered to allow benchmarking against the results of RV44. An additional arm of the study evaluated these vaccines together with the oil-in-water emulsion MF59 adjuvant. Results: We found that alum protected macaques from SIV mac25 acquisition while MF59 did not despite its ability to elicit higher systemic T-cell and antibodies responses. MF59 altered homing of antibody producing cells and increased the frequency of CXCR3 + plasmablasts in blood that positively correlated with anti-envelope IgA serum levels and phagocytosis. Alum, in contrast, increased the frequency of plasmablasts expressing the mucosal integrin a4b7 that positively correlated with IgA responses to cyclic V2 in rectal mucosa. In the alum group mucosal IgG to cyclic V2 correlated with lower risk of SIV mac25 acquisition. However, mucosal IgG to linear and cyclic V2 correlated with an increased risk of SIV mac25 acquisition in the MF59 group. The two adjuvants modulated distinct signaling pathways and ORAL ABSTRACT SESSIONS 09

110 Oral Abstract Sessions Oral Abstract Session 25: Adjuvants and Immunogens OA25.02 IgG Glycosylation Is Programmed and Remembered after Immunization with TLR Stimulating Adjuvants OA25.03 CD4-induced Epitopes Are Exposed on Cell-bound HIV-: The Key to Fc Receptor Mediated Humoral Immunity? Alison E. Mahan, Hamid Mattoo 2, Kendall Dionne, Jacquelynne Tedesco, Shiv Pillai 2, Galit Alter Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 Harvard Medical School, Center for Cancer Research, Charlestown, MA, United States Meron Mengistu, George K. Lewis, Anthony L. DeVico Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, United States ORAL ABSTRACT SESSIONS Background: The N-linked glycan in the CH2 domain of IgG potently affects antibody effector functionality by mediating its affinity for innate immune cell receptors. In particular, glycosylation is important for antibody dependent cellular cytotoxicity (ADCC) and complement deposition, as well as general inflammation. Vaccine correlates analyses support an important role for effector functions, especially ADCC, in protection from HIV infection. Therefore, understanding how to program IgG-glycans through vaccination may prove to be of critical importance. Methods: Mice were immunized with NP-antigen formulated with a variety of adjuvants, including TLR4, TLR5 and TLR7/8 agonists mixed with alum. At 28, 48 and 60 days post immunization, transcriptional profiles of antigen-specific B cells were analyzed using microarray and targeted qpcr. In addition, glycosylation of antigen-specific IgGs was analyzed by capillary electrophoresis. Results: We observed that animals that received TLR7/8 agonist adjuvants produced more inflammatory IgG-glycans and this corresponded to transcriptional levels of important glycosylation enzymes, particularly the galactosylation enzyme B4GALT. Additionally, the effector function modifying sugar fucose and its enzyme transcript, FUT8, were decreased by the TLR5 agonist adjuvant, suggesting that IgG effector functionality can be tuned by specific signals during immunization. Importantly, by boosting some animals at 28 days with antigen alone, we observed that these glycosylation profiles are remembered since these animals maintained the same IgG glycan profile with or without repeated administration of adjuvant. Conclusions: These data are the first to provide evidence that glycosylation can be tuned and remembered after immunization with TLR agonist adjuvants. This is important for understanding how HIV vaccines can be designed to elicit antibodies with good effector function glycan profiles in antigen-specific IgGs. Background: The partially successful RV44 vaccine trial produced non-neutralizing antibodies that mediate ADCC against HIV-. These antibodies recognize the CD4-induced (CD4i) C region of gp20. However, such findings are enigmatic in view of previous arguments that CD4i epitopes are hidden on viral trimers before, and during interaction with host cell. It is therefore critical to understand the antigenicity of these conserved epitopes that become exposed during productive viral replication, to provide important guidance for designing antiviral strategies such as vaccines to raise protective antibody responses. Methods: We studied the antigenicity of conserved epitopes by visualizing their exposure on single HIV JRFL virus particles as they interact with target TZM-bl cells using confocal microscopy. Epitope exposure was probed by visualizing the binding of neutralizing antibodies b2 and 2G2, and CD4i antibodies A32, 7b and C. To reconcile the question of CD4i antigenicity, we examined the location of CD4i antibody attachment with 20nm precision using superresolution microscopy. Results: CD4i antibodies can access their cognate epitopes on TZM-bl cell-bound HIV JRFL virions. Patterns of exposure varied with antibody; however, in general the level of CD4i exposure was similar to neutralizing epitopes b2 and 2G2. CD4i epitopes appear distal to the virus-cell contact site, where they can be accessed by antibodies involved in ADCC. Conclusions: The patterns of CD4i epitope exposure are consistent with the ADCC activities of cognate antibodies against bound virions. CD4i antibodies were able to gain access to their targets due to the unexpected epitope exposure on gp20, distal to the site of contact with cell surface CD4. These findings indicate that HIV- exhibits a diversity of epitope exposure upon attachment that may provide unique insights for understanding how humoral immunity impacts HIV infection. 0 HIV Research for Prevention 204 HIV R4P

111 Oral Abstract Session 25: Adjuvants and Immunogens Thursday, 30 October OA25.04 OA25.05 Structural Evolution of HIV- gp20 Glycan Recognition by the PGT2 Lineage of Potent Broadly Neutralizing Antibodies Fernando Garces, Devin Sok 2, Leopold Kong, Ryan McBride 3, Helen J. Kim 4, Karen F. Saye-Francisco 2, Jean-Philippe Julien, Yuanzi Hua 4, Albert Cupo 5, John P. Moore 5, James C. Paulson 6, Andrew B. Ward, Dennis R. Burton 7, Ian A. Wilson 8 Dept of Integrative Structural & Computational Biol, The Scripps Research Institute, IAVI Neutralizing Antibody Center, Scripps CHAVI- ID, La Jolla, CA, United States, 2 IAVI Neutralizing Antibody Center, Scripps CHAVI-ID, Dept Immunology & Microbial Science, The Scripps Research Institute, La Jolla, CA, United States, 3 Dept of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, CA, United States, 4 Dept of Integrative Structural & Computational Biol, The Scripps Research Institute, La Jolla, CA, United States, 5 Dept of Microbiology & Immunology, Weill Medical College of Cornell University, New York, NY, United States, 6 Dept of Cell and Molecular Biology, The Scripps Reaserch Institute, La Jolla, CA, United States, 7 IAVI Neutralizing Antibody Center, Scripps CHAVI-ID, Dept Immunology & Microbial Science, The Scripps Research Institute, Ragon Institute of MGH, MIT and Harvard, Cambridge, MA 0239, La Jolla, CA, United States, 8 Dept of Integrative Structural & Computational Biol, The Scripps Research Institute, IAVI Neutralizing Antibody Center, Scripps CHAVI-ID, Skaggs Institute for Chemical Biology, La Jolla, CA, United States Background: The HIV- envelope glycoprotein trimer is the sole target of the neutralizing antibody response and, therefore, the prime platform for vaccine design. The PGT2 lineage of antibodies (PGT2-24; PGT33-34) exhibits exceptionally potent and broad neutralizing activity against HIV- at serum concentrations achievable by vaccination. Recently, the crystal structure of PGT22 in complex with BG505 SOSIP.664 gp40 provided a view of how an affinitymatured antibody from the PGT2 family recognizes gp20. PGT24, a bnab from the same family, appears to represent an alternative branch in the antibody maturation process. Therefore, we have an opportunity to investigate the different ways in which high affinity recognition of a complex epitope involving glycans and protein surfaces can evolve from the same germline precursor. Methods: Towards this end, we used X-ray crystallography and EM to unveil the molecular details of the PGT24-gp20 interaction, and deep sequencing, virus neutralization, calorimetry (ITC), and glycan arrays to add additional functional and biochemical insights. Results: The crystal structure of PGT24 in complex with the gp20 at 3.3Å resolution reveals a novel mode of recognition involving primarily the glycan at position N332 and a GDIR protein motif at the V3 loop base. We also discovered that residues on PGT24 that recognize this epitope are conserved throughout the PGT2 lineage and may represent the initial requirement for selection and subsequent evolution of this antibody family. Interestingly, residues on PGT22 that recognize additional protein regions and N-linked glycans appear to evolve separately and are conserved only on one branch of the maturation tree. Conclusions: The PGT2 family shows divergent glycan recognition profiles during antibody maturation, presumably in response to virus escape and sequence evolution. This information should enable the design of better scaffolds and variants of native-like SOSIP trimers leading to a successful vaccine. Refocussing Antibody Responses by Chemical Modification of Vaccine Antigens Torben Schiffner, Karolis Leonavicius 2, Heiko Schuster 2, Helen J. Kim 3, Leopold Kong,3, Regis Saliba 2, Florian Brod, Frank Wegmann, Po-Ssu Huang 4, Guillaume B. Stewart-Jones 5, William R. Schief 3,4,6, Andrew B. Ward 3, John P. Moore 7, Rogier W. Sanders 7,8, Benjamin G. Davis 2, Quentin J. Sattentau University of Oxford, Sir William Dunn School of Pathology, Oxford, United Kingdom, 2 University of Oxford, Department of Chemistry, Oxford, United Kingdom, 3 IAVI Neutralizing Antibody Center at The Scripps Research Institute, San Diego, CA, United States, 4 University of Washington, Washington, WA, United States, 5 University of Oxford, The Weatherall Institute of Molecular Medicine, Oxford, United Kingdom, 6 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 7 Weill Cornell Medical College, New York, NY, United States, 8 University of Amsterdam, Amsterdam, Netherlands Background: The HIV- envelope glycoprotein (Env) has developed several immune-evasion mechanisms to avoid the induction of neutralizing antibodies, including immuno-dominant non-neutralizing epitopes, conformational flexibility of conserved epitopes, and spontaneous subunit dissociation. Here, site-specific immuno-silencing by glycan masking and chemical fixation of native-like Env trimers are explored to overcome these obstacles. Methods: Immunogens, including next-generation BG505 SOSIP.664 trimers, were chemically modified in vitro by either site-directed glycan addition or chemical cross-linking, and effects on antibody recognition were characterized in vitro by ELISA and SPR. Immunogenicity was tested in mice and refocussing of antibody responses was analysed by cross-competition with monoclonal antibodies. Results: In vitro, glycan masking led to selective reduction in binding of antibodies recognizing epitopes containing, or proximal to, modification sites (lysine residues) whereas binding of antibodies to epitopes devoid of lysines remained unchanged. Chemical cross-linking of native-like gp40 trimers led to reduced binding of non-neutralizing antibodies including V3-loop directed antibodies, which further significantly increased the existing differential in antibody binding between neutralizing and non-neutralizing antibodies. Immunization with modified antigens led to reduced immunogenicity of silenced epitopes compared to unmodified controls. In contrast, some epitopes that were unaffected by chemical modification showed significantly increased cross-competition with sera from immunized animals. Conclusions: The chemical modifications developed here resulted in improved antibody recognition profiles in vitro and led to selective refocussing of antibody responses in vivo. Thus, chemical modification of vaccine antigens to stabilize antigen and refocus B cell responses presents an attractive tool for vaccine immunogen design. ORAL ABSTRACT SESSIONS

112 Oral Abstract Sessions Oral Abstract Session 25: Adjuvants and Immunogens OA25.06 LB Native-like BG505 SOSIP.664 Trimers Induce Autologous Tier-2 NAbs against Complex Epitopes in Rabbits and Macaques John P Moore Weill Medical College, New York, NY, United States ORAL ABSTRACT SESSIONS Background: Our goal is to induce bnabs by using soluble, recombinant gp40 mimics of the native Env trimer. At present, the only way to make true trimer mimics is to ensure that the gp40 proteins are cleaved between the gp20 and gp4 subunits, and then stabilized via SOSIP mutations. Our team has described the production and properties of native-like BG505 SOSIP.664 trimers, including their high-resolution X-ray and cryo-em structures. Methods: As no native-like, soluble recombinant trimer has previously been evaluated as an immunogen, we tested the BG505 SOSIP.664 trimers in rabbits and macaques. NAb titers were quantified using the TZM-bl cell assay, and their specificity assessed by several techniques, including the use of competitor ( dump-in ) antigens. Results: The BG505 SOSIP.664 trimers induce strong and consistent NAb responses to the autologous Tier-2 BG505.T332N virus (20/20 rabbits, 3/4 macaques), as well as heterologous Tier- NAb responses. Cross-reactive Tier-2 NAbs were not induced. Comparator, non-native BG505 Env proteins performed much worse, exemplified by an uncleaved gp40 that induced autologous Tier-2 NAbs in 0/4 rabbits. The Tier- and Tier-2 NAb titers were not correlated among the animals, implying that they are entirely different responses. A variety of dumpin competitors were used to characterize the NAb epitopes. Tier-, but not Tier-2, NAbs were substantially depleted by simple V3 peptides. The most prevalent Tier-2 response was influenced by C3 sequences present in escape variants that emerged in the BG505 HIV--infected infant. In one rabbit the key Tier-2 NAb epitope was a composite of the V/V2/ V3 regions. Conclusions: In infected people, bnabs evolve from a narrow-specificity autologous Tier-2 response. To induce the Tier-2 breadth necessary for an effective vaccine, we will explore several approaches including the use of new, native-like SOSIP.664 trimers based on subtype B and C sequences. GMP-grade BG505 trimers are now being developed for human trials. 2 HIV Research for Prevention 204 HIV R4P

113 Thursday, 30 October Oral Abstract Session 26: Microbicides and Multipurpose Prevention Technologies OA26.0 OA26.02 Introduction of the SILCS Diaphragm as a Multipurpose Technology in South Africa: Potential Users, Perceived Benefits, and Barriers to Use Cecilia Milford, Letitia Rambally, Muriel Kubeka, Lizzie Moore, Mags Beksinska, Maggie Kilbourne-Brook 2, Jennifer Smit MatCH Research, Department of Obstetrics and Gynaecology, University of the Witwatersrand, Durban, South Africa, 2 PATH, Seattle, WA, United States Background: SILCS, a single-size diaphragm designed to improve women s options for non-hormonal barrier contraception, is being evaluated as a reusable delivery system for microbicide gel, allowing SILCS to be a multipurpose prevention technology (MPT). In the context of high unplanned pregnancies and HIV prevalence in South Africa (SA), a health systems assessment was conducted to identify opportunities and challenges for future introduction of SILCS as a MPT in SA. Potential future users, perceived benefits, and barriers to use of SILCS were identified and described. Methods: Key informant interviews were held nationally (with regulatory authority, policymakers, healthcare providers, trainers, trialists, advocacy groups; n=3) and 3 focus groups were held with potential users ( male, 2 female; n=24) at a primary healthcare clinic in ethekwini District, KwaZulu-Natal. Discussions were transcribed and translated. Data were coded, results organised according to key themes, and NVivo used to facilitate data analysis. Results: Respondents described the advantage of SILCS as a MPT it protects from so many things. It was seen as a woman-initiated method which can be inserted covertly when condom negotiation is difficult. Participants described multiple potential user types those who want to avoid hormonal contraceptives, find it difficult to access health care, educated/mature women, not afraid to insert vaginal products and interested in birth spacing. Barriers to use included misunderstandings of vaginal anatomy, fit ( Will it not disappear here inside? ), concerns about semen spillage, potentially painful insertion, and partner response during sex. There were questions about SILCS care, durability, and use in multiple rounds of sex. Conclusions: Potential SILCS users include a variety of women, but uptake may be based on individual preferences and needs. Most barriers to use can be addressed via healthcare provider training and education of potential users and their partners. A Simple Intravaginal Ring Pump for Sustained Vaginal Release of ARV Microparticles and Macromolecular Agents Ryan Teller,2, Rachna Rastogi 2, Patrick Kiser Northwestern University, Evanston, IL, United States, 2 University of Utah, Salt Lake City, UT, United States Background: Intravaginal ring technology is generally limited to releasing low molecular weight species that can diffuse through the IVR elastomer. To increase the diversity of the drugs that can be delivered from IVR, we sought to create a controlled delivery technology that could uncouple the mechanism of drug release from the interaction of the antiretroviral with the elastomer and provide near zero order release of any stable molecule. We designed an IVR pump that contains pellets of a mixture of micronized drug and hydroxypropyl cellulose (HPC) that are contained in the hollow core of the IVR. In this system orifices control the hydration rate of the pellet and flux of the drug-containing HPC gel that is forced through the orifice by polymer swelling and which then distributes in the vaginal canal. Methods: Each IVR-pump contained ARV/HPC pellets within polyether urethane tubing containing orifices that were sealed by induction welding. We formulated several of the leading PrEP ARV: TDF, tenofovir, maraviroc, and IQP-0528 and several macromolecules. We evaluated IQP-0528 IVR-pumps in sheep for drug release rate, and drug concentration and distribution in the vaginal canal. Results: Altering the type of swelling polymer, drug loading, orifice design and the mass of pellets can control the drug release rate and duration. Formulations for high molecular weight species like IgG and linear polymers could be obtained that were nearly zero order for a month. Less controlled release profiles were achieved with more watersoluble drugs including maraviroc, tenofovir and TDF since the drugs possess much higher diffusivity in the hydrated polymer matrix. We observed mg per day release rates in sheep with an average IQP-0528 concentration in vaginal fluid of 270 µg/ml (~0 5 x the IC 90 ). Conclusions: This device provides an adaptable platform for vaginal drug delivery. We have been able to deliver impressive quantities of hydrophobic drugs as microparticles and high molecular weight agents for a month in animal models. ORAL ABSTRACT SESSIONS 3

114 Oral Abstract Sessions Oral Abstract Session 26: Microbicides and Multipurpose Prevention Technologies OA26.03 OA26.04 ORAL ABSTRACT SESSIONS Development of Reservoir Vaginal Rings Containing Dapivirine or Hormonal Contraceptive Steroids Clare McCoy, Peter Boyd, Susan Fetherston 2, Ian Major, Diarmaid Murphy, Andrew Brimer 3, Jonathon Holt 3, Brid Devlin 3, Wendy Blanda 3, Tiffany Derrick 3, Chris Gimour 3, Jeremy Nuttall 3, Karl Malcolm Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 University of Ulster, School of Pharmacy and Pharmaceutical Sciences, Coleraine, United Kingdom, 3 International Partnership for Microbicides, Silver Spring, MD, United States Background: By extending the use duration of vaginal rings (VRs), the overall production cost per patient per month can be greatly reduced. Matrix-type ring designs produce an initial burst of drug release, and concentrations during the burst must be maintained within safe limits. This constrains both the maximum drug load and, in turn, the use duration of a matrix ring. By contrast, reservoir-type VRs comprise a drug-loaded reservoir surrounded by a rate-controlling non-medicated sheath, such that initial burst release is minimised and constant daily release rates are achieved. In this way, greater drug loadings and longer use periods are possible. Here, we report the development of reservoir VRs formulations containing dapivirine (DPV) and various contraceptive steroids. Methods: Reservoir VRs were manufactured with injection molded cores containing ethinyl estradiol (EE), etonogestrel (ETO), levonorgestrel (LNG) or DPV. Addition- and condensation-cured silicone cores were tested. All cores were overmolded with a blank.5 mm addition-cured silicone sheath. In-vitro release was assessed for up to 60 days. Results: VRs comprising addition cured silicone cores and blank addition cured silicone sheath layers provided zero order release of DPV (95 µg/ day) and LNG (30 µg/day), but not EE (detectable release on days 2 to 0 only) or ETO (no detectable release at any time). Rings comprising a condensation-cured silicone core provided zero order release of DPV (9 µg/day), EE (328 µg/day, days 4 to 8), ETO (422 µg/day, days 4 to 8) and LNG (69 µg/day). Conclusions: Reservoir-type VRs offering zero order release kinetics have potential as a multipurpose prevention technology (MPT) product for contraception and HIV-prevention with use duration of several months. However, the contraceptive hormones require careful selection of the silicone polymer system to ensure adequate release, while DPV is readily released from both types tested. Engineering a Virus Trap and Safety Net Microbicide Stella E. Aniagyei, Jill M. Steinbach University of Louisville, Bioengineering, Louisville, KY, United States Background: Multipurpose drug delivery systems (MDDS) have the potential to impart a variety of attributes to microbicide delivery. We have previously developed a safe and effective microbicide, using poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) that encapsulate short interfering (sirna) targeting host receptors, to significantly increase survival in a murine model of HSV-2 infection; and have more recently developed surface-modified NPs that enhance cell internalization by 70%. Building upon these abilities in our new lab, we seek to create a proof-of-concept MDDS that integrates some of the protective attributes of mucus, including structure and pathogen adhesion, to target the integral stages of HIV- and HSV-2 infection. To achieve this, our MDDS incorporates 3 components: () a biological lectin, Griffithsin (GRFT), to immobilize and debilitate HSV-2 and HIV upon entry ( trapping ); and safety nets of: (2) polymeric electrospun fibers (EFs) and (3) NPs to provide prolonged release of encapsulated antiviral drugs and sirna, respectively. Methods: EFs were electrospun with different solvents and compositions, containing our model small molecule antiviral, Acyclovir (ACV). A portion of EFs were surface modified with our trapping lectin, GRFT. Virus assays were conducted to test the efficacy of ACV EFs and blank GRFT-EFs against HSV-2 infection in vitro. Results: EFs with a range of diameters were produced, depending on solvent selection and electrospinning conditions. We achieved high ACV encapsulation, with cumulative release spanning 20-60%. EFs encapsulating ACV inhibited HSV-2 infection by >90%, while GRFT-EFs immobilized HSV-2. We are currently expanding this platform to inhibit both HSV-2 and HIV infection. Conclusions: We have complemented our sirna NP and surfacemodified NP microbicide platform, to create novel unmodified and GRFT-EFs for use in a MDDS. We are excited to further develop and combine these technologies to eventually capture, prevent, and treat STIs in one platform. 4 HIV Research for Prevention 204 HIV R4P

115 Thursday, 30 October Oral Abstract Session 26: Microbicides and Multipurpose Prevention Technologies OA26.05 A Single Dose Novel Formulation of Maraviroc Using Electrospun Fabrics Designed for Instant and Multi-day HIV Prevention Cameron Ball, Shih-Feng Chou, Yonghou Jiang, Kim A. Woodrow University of Washington, Bioengineering, Seattle, WA, United States OA26.06 LB Mass Spectrometry Imaging of Hair Strands Allows for Evaluation of Long Term Antiretroviral Adherence Corbin Thompson, Elias Rosen 2, Mark Bokhart 2, Heather Prince, Craig Sykes, David C. Muddiman 2, Angela D.M. Kashuba University of North Carolina at Chapel Hill, Chapel Hill, NC, United States, 2 North Carolina State University, Raleigh, NC, United States Background: Current microbicide formulations fail to provide both instant and multi-day sustained ARV delivery with a single dose. Such products could be particularly useful for delivering maraviroc (MVC), which is required at high doses for efficacy but absorbed rapidly in the vagina. To address this need, we electrospun a composite fabric that provides both a rapid burst and a multi-day sustained release of MVC. Fully tunable sustained release is achieved via novel core-shell fiber architectures with high drug loading, and layering with rapid-release fibers allows independent control over the ratio of burst to sustained release. Methods: MVC was formulated into coaxially electrospun fibers with core-shell nano-architectures for sustained delivery. Release of MVC was measured under sink conditions with multiple release media. We probed the mechanism for sustained release from core-shell fibers by measuring material physical and chemical properties, hydration, and mass loss. Finally, core-shell fabrics were combined with rapidly dissolving MVC fibers using either alcohol welding or physical pressure to create soft, single dose fabrics for both rapid and sustained MVC release. Results: Core-shell fibers were loaded with up to 37.5 wt% MVC and provided near linear release for 5 days. Adjusting the drug loading and the ratio of shell-to-core content in fibers provided direct control over the dose and timing of drug release, respectively. The primary mechanism for sustained drug delivery from core-shell fibers, unlike core-shell IVRs, was controlled surface wetting. Combining rapid and sustained delivery fabrics into single doses provided full control over both pericoital (20 min) and sustained (5 days) release. Conclusions: These electrospun MVC composites are the first microbicides to realize fully adjustable biphasic release with a single dose. Translatability to other drugs and ease of manufacturing scale up make this microbicide platform suitable for simultaneous rapid and sustained multipurpose prevention. Background: Successful pre-exposure prophylaxis requires antiretroviral (ARV) adherence. Therapeutic drug monitoring provides accurate, but short term adherence data. Hair strand (HS) analysis with traditional LC/ MS provides long term adherence information, but fails to discriminate ARV presence at the time of HIV exposure. Mass spectrometry imaging (MSI) offers the ability to visualize ARV exposure in tissues, allowing for identification of distinct distribution patterns. Here, as proof of concept, we use MSI to visualize tenofovir (TFV), emtricitabine (FTC), and efavirenz (EFV) in HS from HIV positive subjects. Methods: HS (20-30) were taken from 5 HIV positive, virologically suppressed subjects receiving Atripla for > year. HS incubated in TFV, FTC, and EFV for 24 hours or HS never exposed to ARVs served as positive and negative controls, respectively. After collection, HS were analyzed using an infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) source coupled to a Thermo Q-Exactive mass spectrometer. Signal intensity between HS from a single subject was compared to assess intra-subject variability. MSI data were analyzed using MSiReader software. Results: MSI experiments demonstrate continuous EFV signal along HS for all 5 subjects. MS/MS analysis and the use of a negative control confirmed EFV specificity. Variability in signal intensity was similar within (~ 2.5 fold) and between (~ 4 fold) subjects. TFV and FTC were not visualized in HS from dosed subjects, though FTC and EFV were both detected in the positive control sample. Conclusions: EFV distribution in HS demonstrates consistent ARV exposure in these subjects, and is in agreement with their suppressed viral loads. Confirmation of EFV signal with MS/MS and low intra-subject variability showcase the accuracy and precision of this method. We show for the first time that MSI of HS is a promising approach for measuring ARV adherence. Future studies will examine ARV hair distribution patterns in non-suppressed patients. ORAL ABSTRACT SESSIONS 5

116 Oral Abstract Sessions Oral Abstract Session 27: PrEP: Self-testing, Safety and Modeling OA27.0 Selection of Rilpivirine Resistant HIV- in a Seroconverter on Long-acting Rilpivirine (TMC278LA) from the Lowest Dose Arm of the SSAT 040 Trial Kerri J. Penrose, Urvi M. Parikh, Kristen A. Hamanishi, Constantinos Panousis, Laura Else 2, David Back 2, Marta Boffito 3, Akil Jackson 3, John W. Mellors University of Pittsburgh, Pittsburgh, PA, United States, 2 University of Liverpool, Liverpool, United Kingdom, 3 SSAT Research, Chelsea & Westminster Hospital, London, United Kingdom OA27.02 Uptake of HIV Self-testing among People Receiving PrEP in Kenya Kenneth Ngure,2, Renee Heffron 2, Nelly Mugo 2,3, Elizabeth Irungu 4, Njambi Njuguna 4, Lawrence Mwaniki 4, Kerry Thompson 5, Connie Celum 2,6,7, Jared M. Baeten 2,6,7 Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya, 2 University of Washington, Global Health, Seattle, WA, United States, 3 Kenya Medical Research Institute, Center for Clinical Research, Nairobi, Kenya, 4 Partners in Health Research and Development, Thika, Kenya, 5 University of Washington, Seattle, WA, United States, 6 University of Washington, Epidemiology, Seattle, WA, United States, 7 University of Washington, Medicine, Seattle, WA, United States ORAL ABSTRACT SESSIONS Background: Rilpivirine (RPV) is a potent second-generation NNRTI whose injectable long-acting formulation (TMC278LA) is a candidate for PrEP. A participant in the lowest dose arm of a single dose TMC278LA PK study unexpectedly seroconverted 84 days (d) post-injection. We evaluated plasma samples for residual RPV and HIV- drug resistance after seroconversion. Methods: SSAT 040 evaluated plasma PK of TMC278LA following a single IM dose of 300, 600 or 200mg in 60 low-risk HIV-negative women. RPV plasma concentrations were assessed post-injection at regular intervals. Plasma HIV- RNA levels (VL) and drug resistance by standard genotype and allele-specific PCR were determined retrospectively in the participant who seroconverted. Recombinant infectious virus containing full-length RT sequences from plasma was tested for drug susceptibility in TZM-bl cells. Results: HIV- infection occurred in of 20 participants in the 300mg arm. Post-seroconversion testing illustrates selection of K0E after infection: d84 (seroconversion) VL 75,060 copies/ml, < 0.% K0E, 7.5ng/ml plasma RPV; d5 (TDF/FTC/DRV/r initiation) VL 664,925 copies/ml, 9% K0E, 4.ng/ml plasma RPV; d5 VL 6,204 copies/ ml, K0EK mixture, 3.8ng/ml plasma RPV; d99 VL 3,558 copies/ ml, < 0.% K0E, 6.0 ng/ml plasma RPV. Plasma-derived HIV- clones (d5) containing K0E had 4-fold decrease in susceptibility to RPV, compared to d5 clones without K0E, and exhibited cross-resistance to ETR (4-fold), NVP (8-fold), and EFV (4-fold). Conclusions: A 300mg single dose of TMC278LA did not prevent HIV- infection in one participant in the SSAT 040 trial, but sustained low levels of RPV (less than the protein adjusted IC 90 ) post-infection selected RPV resistant HIV-. This is a unique instance of well-documented infection with wild-type HIV- and subsequent selection of resistant virus by persistent drug exposure post-infection. Evaluation of different dose regimens of TMC278LA is ongoing to optimize drug levels for its use as a PrEP agent. Background: Implementation of pre-exposure prophylaxis (PrEP) by HIV uninfected people will require routine HIV testing to minimize PrEP use during acute HIV infection and the potential development of antiretroviral resistance. HIV self-testing could provide an efficient method to increase testing frequency with minimal increase in cost and participant burden. Methods: In the Partners Demonstration Project, an open-label study of PrEP for HIV prevention among high risk HIV discordant couples, HIV testing occurs and PrEP prescriptions are filled at quarterly clinic visits. In this sub-study at the Thika, Kenya site, HIV self-testing training and kits are offered to HIV uninfected participants using PrEP for use during the two months between each quarterly clinic visit. Results: As of April 204, 20 HIV uninfected partners (93 females and 27 males) have enrolled in the HIV self-testing sub-study. The median age is 30 years (interquartile range, IQR: 26-35) and number of years of education is 0 (IQR: 8-2). Among the 58 participants with at least one follow-up visit to date, 89.7% reported conducting HIV self-testing at least once and 92.3% of these reported that using the self-test kit was easy or very easy.7.3% of participants reported that they selftested when they opened a new bottle of PrEP but the majority (78.9%) reported that self-testing did not coincide with a specific event. The majority (57.7%) of participants reported testing alone while 38.5% of participants reported that their study partner was with them. More than half (59.5%) of participants reported sharing the self-test results with the study partner while 36.5% did not share the results with anyone. All self-test results were HIV negative. Conclusions: Within a PrEP demonstration project conducted in a peri-urban African setting, HIV uninfected partners successfully used HIV self-testing with ease. Self-testing may be a feasible, cost effective, adjunctive method to increase the frequency of HIV testing in conjunction with PrEP use. 6 HIV Research for Prevention 204 HIV R4P

117 Oral Abstract Session 27: PrEP: Self-testing, Safety and Modeling Thursday, 30 October OA27.03 The Better it Is, the More it Will Be Used - The Synergistic Relationship between Product Efficacy, Level of Uptake and Overall Impact Fern Terris-Prestholt, Matt Quaife, Sinead Delany-Moretlwe 2, Helen Rees 2, Charlotte Watts, Peter Vickerman,3 London School of Hygiene & Tropical Medicine, Global Health and Development, London, United Kingdom, 2 Wits Reproductive Health and HIV Institute (WRHI), Johannesburg, South Africa, 3 University of Bristol, School of Social and Community Medicine, Bristol, United Kingdom OA27.04 Calcutta HIV Model Projections and the Impact of PrEP Zindoga Mukandavire, Kate Mitchell, Smarajit Jana 2, Peter Vickerman,3 London School of Hygiene and Tropical Medicine, Social and Mathematical Epidemiology Group, London, United Kingdom, 2 Sonagachi Research & Training Institute, Kolkata, India, 3 University of Bristol, School of Social and Community Medicine, Bristol, United Kingdom Background: Mathematical models show the important role product efficacy, user uptake and adherence play in the impact of new HIV prevention technologies (NPTs). However, these models rely on uptake assumptions that at best are based on expert opinion, levels of use in trials or of existing technologies, and none are able to consider how uptake may be implicitly related to how good the product is. Methods: This study uses predicted product uptake from a choice experiment (CE) in South Africa to model microbicide effectiveness: average population protection provided by microbicides with different efficacies. It incorporates differential uptake of products with different efficacies and the degree to which condom users say they will switch to the microbicide. We compare with projections assuming 30% uniform uptake among non-condom users only. Results: For a population of women with 20% condom use at baseline, we predict introduction of a moderately effective microbicide (55% ) will only increase the population protection provided over an average sex act from 7% to 23%, with 6% and % uptake among women who do not and do use condoms, respectively. For 60% condom use average protection is 53%, only.5% higher than without microbicides. The uniform uptake model predicts 30% average protection for a moderately effective microbicide at low condom use and 58% protection for the 60% condom use scenario. If the product is 95% efficacious, added microbicide protection is 3% at low condom use versus 7% at high condom use. Here the uniform uptake model predicts 23% and % added protection, respectively. Conclusions: Microbicide impact assuming uniform uptake is likely to be overestimated for moderately effective products while underestimated for highly effective products. Ignoring this differential effect is likely to lead to biased models and inefficient allocation of resources. In the absence of observed uptake, this study proposes the use of hypothetical CE data to strengthen our impact models. Background: Despite low national HIV prevalence in India, HIV prevalence remains high among high risk groups. We assessed the potential impact of pre-exposure prophylaxis (PrEP) use for HIV prevention among female sex workers (FSWs) in Calcutta, India. Methods: A mathematical model was developed to simulate the transmission of HIV among FSWs and their commercial clients in Calcutta. We parameterised the model using data from Calcutta, with any data gaps being filled with data from Bangalore or Mumbai. The model was fit to FSW HIV prevalence from Calcutta for 992 to 200. The model was analysed to estimate the efficiency and impact of PrEP among FSWs assuming PrEP use starts in 204. We considered different efficacy and coverage scenarios for PrEP. Results: Our model projections suggest that with existing interventions, FSW HIV prevalence is on the decline in Calcutta. The different PrEP intervention scenarios suggest that PrEP use could result in rapid and substantial decreases in HIV prevalence and incidence compared to the baseline scenario of no PrEP, with a 60% reduction in HIV incidence achievable over 20 years with a PrEP efficacy of 60% and coverage of 60%. However, the efficiency of PrEP is low with about 40 years of PrEP being required per life year gained in the most optimistic PrEP scenario (60% coverage and efficacy). Conclusions: PrEP interventions could be important for controlling HIV in FSW driven epidemics in Calcutta and India in general, but may not be cost-effective in many scenarios. ORAL ABSTRACT SESSIONS 7

118 Oral Abstract Sessions Oral Abstract Session 27: PrEP: Self-testing, Safety and Modeling OA27.05 The Potential Impact of Long-acting PrEP on HIV Transmission, Mortality and Drug Resistance in KwaZulu-Natal, South Africa: Model-based Analyses OA27.06 LB A Phase Open Label Safety, Acceptability, Pharmacokinetic, and Pharmacodynamic Study of Intramuscular TMC278 LA (the MWRI-0 Study) ORAL ABSTRACT SESSIONS Robert Glaubius, Greg Hood 2, Ume Abbas Cleveland Clinic, Infectious Disease and Quantitative Health Sciences, Cleveland, OH, United States, 2 Pittsburgh Supercomputing Center, Pittsburgh, PA, United States Background: Long-acting injectable rilpivirine (RPV) pre-exposure prophylaxis (PrEP) could be pivotal for optimizing PrEP effectiveness for HIV prevention. The impact of RPV PrEP on HIV transmission, mortality and the spread of drug resistance are unknown. Methods: We examine the scale-up of RPV PrEP, ART and male medical circumcision (MMC) using a mathematical model of the HIV epidemic in KwaZulu-Natal (KZN). We compare a baseline scenario of ART and MMC scaled up to 80% coverage by 207 to three scenarios of ART and MMC plus ten years of PrEP (90% efficacy; 35% cross-resistance) rollout starting in 205: a) 0% overall coverage of susceptible adults (20% of KZN s HIV budget at $65 per person-year of PrEP), either unprioritized or b) prioritized to women aged 20-29; or c) 80% coverage of high-risk adults (< 2% of the HIV budget). Outcomes include infections averted and drug resistance prevalence over ten years, and lifetime life-years (LY) lived, cost and cost-effectiveness. We classify scenarios as very cost-effective if the incremental cost per LY saved is less than South Africa s per capita GDP of $7,500, and costsaving if costs decrease while life-years increase. Results: Unprioritized PrEP averts 6.8% of infections over ten years and saves.9 million LY in the PrEP-exposed cohort ($605/LY saved). Age-prioritization improves PrEP s impact, averting 2.6% of infections and saving 3.5 million LY ($3/LY saved). Risk-prioritized PrEP reduces costs by 0.7%; it averts more infections (8.4%) and saves more LY (2.3 million) than unprioritized PrEP while using < 0% as many person-years of PrEP. Drug resistance decreases with unprioritized or age-prioritized PrEP (%;.7%), and increases by 2.5% with risk-prioritized PrEP compared to baseline (358,000 cases at 2025). Conclusions: RPV PrEP is potentially very cost-effective, and may be cost-saving when prioritized to high-risk persons. Drug resistance from PrEP is limited compared to ART, though risk-prioritized PrEP may increase overall resistance. Ian McGowan, Aaron Siegel 2, Kathy Duffill 2, Cory Shetler 2, Charlene Dezzutti, Nicola Richardson-Harman 3, Kaleab Abebe, David Back 4, Laura Else 4, Amy Herrick 5, Peter Williams 6, Khaja K Rehman 2, Ross D. Cranston University of Pittsburgh School of Medicine, Pittsburgh, PA, United States, 2 Magee Women Research Institute, Pittsburgh, PA, United States, 3 Alpha StatConsult LLC, Damascus, MD, United States, 4 University of Liverpool, Liverpool, United Kingdom, 5 University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA, United States, 6 Janssen Research and Development, Beerse, Belgium Background: Long acting (LA) injectable antiretroviral agents are being evaluated as a potential strategy for pre-exposure prophylaxis of HIV infection. This study was undertaken to characterize the safety, acceptability, pharmacokinetic (PK), and pharmacodynamic (PD) profile of TMC278 LA. Methods: Healthy HIV- seronegative participants were enrolled into 2 cohorts. 2 women and 6 men received an intramuscular dose of either 200 mg (Cohort ; N=8) or 600 mg (Cohort 2: N=8) of TMC278 LA. Plasma and tissue (rectal [RT], cervical [CT], and vaginal [VT]) were collected before and after exposure to TMC278 LA. Participants were followed for 4 months after receiving TMC278 LA. Safety, acceptability, multicompartmental PK, and PD (ex-vivo explant challenge with HIV- BaL ) were monitored throughout the study. Results: Thirty six participants were enrolled. There were 83 adverse events of which 79 (97.8%) were Grade /2. Two Grade 3 and 2 Grade 4 events were unrelated to study product. The mean acceptability score was 7 out of a maximum of 0. Geometric mean (GM; 90% CI) plasma TMC278 concentrations at Day 28 post-dose for the 200 mg and 600 mg cohorts were 53 (38-67) ng/ml (female) / 43 (23-63) ng/ml (male) and 28 (9-37) ng/ml (female) / 7 (9-24) ng/ml (male) respectively. For the 200 mg dose the GM tissue: plasma ratios at Day 28 were 0.68 (VT), 0.77 (CT),.2 (female RT), and.27 (male RT). Exposure to TMC278 LA was associated with significant suppression of viral replication in RT (p < 0.000) that persisted for up to four months following exposure to TMC278 LA, but viral suppression was not seen in VT or CT. Conclusions: Single dose administration of TMC278 LA was safe and well tolerated with dose dependent plasma PK exposure. The TMC278 tissue: plasma ratio for RT was approximately two-fold higher than VT or CT. This is the first study to demonstrate prolonged suppression of explant viral replication following systemic antiretroviral exposure. Multiple dosing studies of TMC278 LA are planned. 8 HIV Research for Prevention 204 HIV R4P

119 Oral Abstract Session 28: Treating and Preventing: the Role of ARVs Thursday, 30 October OA28.0 High Initiation and Adherence among High Risk African HIV Discordant Couples in a Demonstration Project of PrEP and ART for HIV Prevention Renee Heffron, Connie Celum, Nelly Mugo 2, Elly Katabira 3, Elizabeth Bukusi 2, Stephen Asiimwe 4, Kenneth Ngure 5, Nulu Bulya 3, Josephine Odoyo 2, Edna Tindimwebwa 4, Deborah Donnell 6, Jessica E. Haberer 7, Lara Kidoguchi, Jennifer Morton, Jared M. Baeten, for the Partners Demonstration Project Team University of Washington, Global Health, Seattle, WA, United States, 2 Kenya Medical Research Institute, Nairobi, Kenya, 3 Makerere University, Kampala, Uganda, 4 Kabwohe Clinical Research Center (KCRC), Kabwohe, Uganda, 5 Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya, 6 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 7 Massachusetts General Hospital, Boston, MA, United States OA28.02 Evaluation of a Risk Score Tool to Identify Higher-risk HIV- Serodiscordant Couples for Antiretroviral-based HIV- Prevention Elizabeth M. Irungu, Renee Heffron 2, Nelly Mugo 2,3, Kenneth Ngure 2,4, Elly Katabira 5, Nulu Bulya 5, Elizabeth Bukusi 2,3, Josephine Odoyo 3, Stephen Asiimwe 6, Edna Tindimwebwa 6, Deborah Donnell 2,7, Connie Celum 2, Jared M. Baeten 2, Partners Demonstration Project Team Kenyatta National Hospital, Nairobi, Kenya, 2 University of Washington, Seattle, WA, United States, 3 Kenya Medical Research Institute, Nairobi, Kenya, 4 Jomo Kenyatta University of Agriculture and Technology, Ruiru, Kenya, 5 Makerere University, Kampala, Uganda, 6 Kabwohe Clinical Research Center (KCRC), Kabwohe, Uganda, 7 Fred Hutchinson Cancer Research Center, Seattle, WA, United States Background: Pre-exposure prophylaxis (PrEP) and antiretroviral therapy (ART) have high efficacy for HIV prevention among heterosexual African HIV discordant couples. Assessing initiation and adherence to antiretroviral-based HIV prevention strategies outside of clinical trial settings is a priority. Methods: Enrollment of high-risk HIV discordant couples into an openlabel PrEP and ART demonstration project in Kenya and Uganda began in November 202. Without PrEP and ART, anticipated HIV incidence in this cohort would be 5% per year. ART is offered per national guidelines and PrEP is offered as a bridge to ART, provided between study enrollment and the time when the HIV-infected partner is likely to have achieved viral suppression. ART adherence is assessed through biannual HIV RNA measurements and MEMS caps are used to assess PrEP adherence. Results: As of April 204, 74 high risk couples were enrolled, 67% with an HIV infected woman, and 42% with an HIV infected partner eligible to initiate ART. At enrollment, 96% of HIV uninfected participants initiated PrEP and 92% of ART-eligible participants planned to start ART. To date, 5% of couples have had 6 months of expected follow-up and retention through 6 months is 88% for HIV uninfected and 93% for infected participants. Among HIV uninfected participants on PrEP, 77% took 80% of expected PrEP doses by MEMS between enrollment and their 6 month clinic visit. Travel and relationship dissolution are common reasons for missing PrEP doses, reported at 27% and 8% of visits with missed doses. Among HIV infected participants initiating ART at enrollment, 85% had a plasma HIV RNA concentration < 400 copies/ ml by the 6 month visit. Six months after enrollment, 85% of couples were using PrEP and/or ART: 45% PrEP only, 4% ART only, and 26% both PrEP and ART. Conclusions: PrEP and ART initiation and early adherence are high among high-risk African HIV discordant couples participating in an open-label demonstration project of PrEP and ART for HIV prevention. Background: Antiretroviral therapy (ART) and pre-exposure prophylaxis (PrEP) significantly reduce HIV transmission within heterosexual HIV serodiscordant couples. To maximize impact and minimize costs, ART and PrEP interventions should be offered to couples at highest risk of HIV transmission. Methods: The Partners Demonstration Project is an open-label, prospective cohort study of PrEP and ART for HIV prevention among high risk HIV serodiscordant couples in Kenya and Uganda. We evaluated the feasibility of using a validated empiric risk score (Kahle et al JAIDS 203) that uses a combination of variables (age, number of children, male circumcision status, plasma HIV levels, condom use, marital status) to identify couples at highest risk for HIV transmission. Results: Since November 202, 27 couples have screened and 74 enrolled. Of the screened couples, 274 (23%) scored 0-4 (anticipated HIV incidence of 2% per year), 493 (4%) scored 5-6 (anticipated HIV incidence of ~5% per year) and 450 (37%) scored 7 (anticipated HIV incidence of 7% per year). Couples scoring 5 were eligible for enrollment and 76% entered the study. The median age of the HIV uninfected partner was 29.5 [IQR 26, 36] years and the HIV seronegative partner was male in 67% of partnerships. Over half (57%) had no children, 92% had unprotected sex in the month prior to screening and 34% of HIV susceptible men were uncircumcised. Among HIV infected members of the couples, the median CD4 count was 436 [IQR 274,634] cells/mm 3 and 4.3% had a plasma viral load >50,000 copies/ml. Conclusions: An empiric risk score derived from observational analyses of African HIV serodiscordant couples identified higher risk HIV discordant couples for a demonstration project of PrEP and ART for HIV prevention. Three-quarters of the higher risk couples who were eligible have enrolled. Risk scores could be useful for recruitment of higher risk persons and couples into future HIV prevention trials and demonstration projects. ORAL ABSTRACT SESSIONS 9

120 Oral Abstract Sessions Oral Abstract Session 28: Treating and Preventing: the Role of ARVs OA28.03 OA28.04 ORAL ABSTRACT SESSIONS Facing the Realities of HIV Universal Test and Treat: Early Lessons Learnt from the Delivery of the HPTN07/ PopART Study Intervention in Zambia Kwame Shanaube, Sam Griffith 2, Musonda Simwinga, Ephraim Sakala, Chepela Ngulube, Sandra Simbeza, Martin Mutonyi, Sarah Fidler 3, Richard Hayes 4, Helen Ayles,5, HPTN07-PopART study Team ZAMBART Project, Lusaka, Zambia, 2 FHI 360, Science Facilitation Department, Washington, NC, United States, 3 Imperial College, Department of Infectious Disease Epidemiology, London, United Kingdom, 4 London School of Hygiene & Tropical Medicine, Department of Infectious Disease Epidemiology, London, United Kingdom, 5 London School of Hygiene & Tropical Medicine, Department of Clinical Research, London, United Kingdom Background: The HPTN07 (PopART) is a 5-year community randomized study of a combination HIV prevention package in 2 communities in Zambia and South Africa. HPTN07 is one of the first studies to evaluate a combined HIV prevention package (including universal HIV testing and ART for all HIV+ individuals irrespective of CD4 count (UTT)) on HIV incidence at community-level. Methods: 2 communities were randomly assigned to one of three study arms. The study intervention consists of door to door voluntary UTT with the offer of immediate ART to all individuals who test positive, plus other HIV preventive methods. Community HIV care providers (CHiPs) are responsible for the delivery of the intervention and linkage to care. Multi-disciplinary groups have been closely engaged in designing and developing the study protocol and include community representatives, community advisory boards, policy makers, government, ministries of health, care and service providers, funders and research teams. Results: Trial initiation was preceded by two years of preparatory work. Early rapid formative research to identify local catalysts and barriers that could influence intervention proved invaluable, as have regular door to door and community mobilization activities. Study preparation required strengthening existing health care systems and improvement of supply chain management. Hiring and training of approximately 430 community based staff to deliver the intervention was done. Mechanisms to best deliver some clinic activities-routinely and in optimal time- are still being identified. Regular feedback on progress and challenges at all levels through sharing of process data is useful. Synchronization with other ongoing HIV preventive initiatives in these communities has been occurring. Conclusions: Initiating a complex combination-prevention trial at a population level, that is integrated within current DoH facilities and therefore sustainable, requires multi-disciplinary approaches, time and effort. HIV Transmission in Discordant Couples in Non Research Settings: Rwanda Experience Etienne Karita, Rachel Parker 2, Sabin Nsanzimana 3, Placidie Mugwaneza 3, Roger Bayingana, Robertine Sinabamenye, Gisele Umvirigihozo, Nuri Ahmed, Kristin Wall 4, Amanda Tichacek 2, Eric Hunter 2, Susan Allen 2 Project San Francisco, Kigali, Rwanda, 2 Emory University, School of Medicine, Atlanta, GA, United States, 3 Rwanda Biomedical Center, Kigali, Rwanda, 4 Emory University, Rollins School of Public Health, Atlanta, GA, United States Background: Most new HIV infections in sub-saharan Africa occur through heterosexual transmission in stable couples. The HPTN 052 trial provided evidence that antiretroviral treatment (ART) significantly reduces the risk of transmission among discordant couples (DC). However, little is known of its effectiveness in the real world. The objective of our study was to measure the risk of HIV transmission among DC followed in public health centers (PHC) in Kigali, Rwanda. Methods: The Rwandan Ministry of Health has established Couples Voluntary Counseling and Testing (CVCT) for HIV as a national policy in antenatal care. Since November 20, Project San Francisco (PSF) is providing mentorship to 20 high volume PHC in Kigali for the follow-up of DC. In this program, HIV-infected partners are offered ART, and HIVnegative partners are offered risk reduction counseling and repeat HIV testing on a quarterly basis. Results: From November 20 to March 204, 3025 DC were registered in the 20 PHC. Of these, the index partner in 367 (45%) were on ARV. 827 (60%) couples (070 M+F- and 757 M-F+) had at least one followup visit, of whom 940 were on ARV at enrollment and 467 initiated ART during follow-up. During follow-up, 39 previously HIV- women and 0 men seroconverted. Viral linkage analysis was performed on 6 out of the 20 transmission pairs in whom the index partner was on ART, and 8% were linked. The overall HIV incidence rate was 0.7 per 00 person-years (95% CI: 0.4,.3) when the index partner was on ARV, and 5.33 (95% CI: 3.54, 7.70) when the index partner was off ARV. Conclusions: Our program data show that in the real world if CVCT and routine follow-up are well established and ART can be offered, ART is associated with an 87% reduction in the risk of HIV transmission from discordant partnerships. However, as transmission from treated index partners is being observed, ongoing risk reduction and ART adherence counseling should be re-emphasized in Treatment as Prevention programs. 20 HIV Research for Prevention 204 HIV R4P

121 Oral Abstract Session 28: Treating and Preventing: the Role of ARVs OA28.05 Mathematical Modelling to Estimate the Impact and Cost-effectiveness of TasP, PrEP and Condom Promotion for Serodiscordant Couples in Nigeria Thursday, 30 October Kate M. Mitchell, Aurélia Lépine, Fern Terris-Prestholt, Emmanuel Alhassan 2, John Idoko 2, Peter Vickerman,3 London School of Hygiene and Tropical Medicine, Global Health and Development, London, United Kingdom, 2 National Agency for the Control of AIDS, Abuja, Nigeria, 3 University of Bristol, Bristol, United Kingdom Background: In Nigeria, only 35% of those in need receive antiretroviral therapy (ART). We used mathematical modelling to estimate the impact and cost-effectiveness of treatment as prevention (TasP), pre-exposure prophylaxis (PrEP) and condom promotion for discordant couples in Nigeria. Methods: A deterministic model of HIV transmission within discordant partnerships and to/from external partners was parameterised using data from Nigeria and other African settings. The impact (infections averted (IA) and disability-adjusted life-years (DALYs) averted) and incremental cost-effectiveness ratios (ICER) were estimated for offering condom promotion, PrEP to HIV-negatives and/or TasP to HIV-positives, compared with a baseline scenario where ART was offered at current national guidelines (CD4< 350 cells/µl) to all HIV positive partners. Full costs (US$202) were estimated from a provider perspective, with future costs and impacts discounted. Frontier analysis was used to determine optimal intervention combinations. Results: Large benefits came from offering ART to all HIV positive partners at current national guidelines compared with current ART coverage (36 IA (23% of infections), 600 DALYs averted over the lifespans of 000 couples and their external partners). Condom promotion was the most cost-effective strategy to provide after offering ART at national guidelines (US$650/DALY), with the addition of TasP with condom promotion being the next best investment (ICER US$050/DALY). The addition of PrEP was not cost-effective, but provided substantial additional impact in terms of IA (an additional 7 IA when added to TasP + condom promotion). Conclusions: The best first intervention for discordant couples in Nigeria would be offering ART at current national guidelines to all HIV-positives. Additional impact could be efficiently achieved from condom promotion and TasP for HIV-positives. PrEP would avert additional infections, but is not cost-effective in terms of DALYs averted once other interventions are in place. ORAL ABSTRACT SESSIONS 2

122 Oral Abstract Sessions Oral Abstract Session 29: T Cell Immunity OA29.0 OA29.02 ORAL ABSTRACT SESSIONS Dendritic Cells from HIV- Neutralizers Efficiently Induce the Generation of CXCR5 + CXCR3 + PD Lo CD4 T Cells with B Cell Helper Function Enrique Martin-Gayo, Taylor Hickman, Zhengyu Ouyang, Rafael Cubas 2, Madelene Lindqvist, Daniel Kauffman, Elias Haddad 2, Bruce Walker,3, Mathias Lichterfeld 4, Xu G. Yu Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States, 2 Vaccine and Gene Therapy Institute (VGTI) of Florida, Port Saint Lucie, FL, United States, 3 Howard Hughes Medical Institute (HHMI), Boston, MA, United States, 4 Infectious Disease Division, Massachusetts General Hospital, Boston, MA, United States Background: Cellular mechanisms supporting the evolution of broadly neutralizing antibody activity (bnabs) against HIV- are poorly understood. Here, we investigated if conventional dendritic cells (cdc) from HIV- patients who naturally develop bnabs have superior ability to prime naïve CD4 T cells into Follicular helper (TFH)-like cells. Methods: cdcs isolated from healthy persons, HIV-+ chronic progressors (CP) and controllers with and without detectable bnabs in plasma (Neutralizers and Non Neutralizers, respectively) were co-cultured with identical allogeneic naïve B cells and T cells. TFH phenotypic and functional characteristics of primed T cells were compared to peripheral TFH-like cells isolated from human blood. Results: cdcs from all patient cohorts similarly induced the generation of PD hi CXCR3 + TFH-like cells. However cdcs from Neutralizers promoted more efficiently the development of PD lo CXCR3 + TFH-like cells and subsequent maturation of B cells. Both PD hi and PD lo cells expressed TFH markers such as ICOS, Bcl6 and CXCR5, but PD lo cells preferentially expressed central-memory and T memory stem cells markers, and had improved abilities to survive in vitro, as opposed to PD hi cells that were highly susceptible to apoptosis and exhibited an effector-memory phenotype. Interestingly, upon Ag stimulation, PD- lo TFH-like cells isolated from peripheral blood differentiated into PD hi TFH-like cells. Importantly, PD lo and PD hi TFH-like cells were both detectable in peripheral blood and lymphoid tissues, but higher proportions of PD lo TFH-like cells were found in the blood of HIV controllers who developed neutralizing breadth. Conclusions: cdcs from Neutralizers have increased abilities to prime the generation of PD lo TFH-like cells, which serve as progenitors of effector PD hi TFH-like cells, but also provide help to B cells. Higher proportions of PD lo TFH-like cells in the blood of HIV- neutralizers suggest an important role of these cells for supporting the development of bnabs Massive Activation, Expansion and Subsequent Apoptosis of CD8 T-cells in Acute HIV Infection Zaza Ndhlovu,2, Philomena Kamya 2, Nikoshia Mewalal, Thandeka Nkosi, Nasreen Ismail, Amber Moodley, Krista Dong, Thumbi Ndung u, Bruce Walker 2 University of KwaZulu-Natal, HIV Pathogenesis Programme, Durban, South Africa, 2 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States Background: The initial CD8 T cell response following acute HIV infection suppresses virus replication, but viremia persists in the vast majority of people. We used pre-infection and very early HIV infection longitudinal samples (ranging from day to day 60 post Fiebig I) to interrogate the dynamics and fate of the initial T cell response, during the initial rise and subsequent decline in plasma viremia. Methods: High risk uninfected women in KwaZulu Natal, South Africa were screened twice weekly by HIV RNA, as part of a comprehensive HIV prevention program. Pre-infection and post infection blood samples were evaluation by intracellular cytokine staining (Cd4, CD8, Ki67, Bcl- 2, CD38, HLA-DR) as well as IFN-gamma ELIspot, and class I tetramer staining assays. Results: Acute HIV infection rapidly induced massive activation and expansion of HIV-specific CD8 T cells such that more than 90% became activated by day 4 days post Fiebig I, upregulating CD38, HLA-DR, and Ki67. In contrast, Influenza and CMV-specific CD8 T cells were not activated. At peak activation HIV-specific CD8 T cells selectively expressed high levels of the pro-apoptotic marker CD95, low antiapoptotic molecule BCL-2 and failed to upregulate the IL-7 receptor. Overnight in vitro incubation of peak activation CD8 T cells resulted in high levels of spontaneous cell death compared to cells collected after the resolution of T cell activation. Furthermore, activated CD8 T cells were also less responsive to ex-vivo stimulation compared to samples collected after resolution of activation, suggesting maximal in vivo activation. Conclusions: These data indicate that primary HIV infection induces massive CD8 T cell expansion, which declines despite persistence of detectable viremia. The disappearance of early responses may be attributed to persistent elevation of antigen-stimulation driving virusspecific cells towards apoptosis. Therefore strategies aimed at blocking apoptosis might be a viable option for rescuing or strengthening early responses. 22 HIV Research for Prevention 204 HIV R4P

123 Oral Abstract Session 29: T Cell Immunity OA29.03 Thursday, 30 October OA29.04 Reduction in Breadth and Not Polyfunctionality or Proliferative Capacity of CD8+ T Cells Is Associated with Loss of Virologic HIV Control Catherine K. Koofhethile, Zaza Ndlhovu,2, Nasreen Ismail, Zanele Mncube, Lungi Maphumulo, Mary van de Stok, Christina Thobakgale, Julia G. Prado 3, Bruce B.D. Walker,2, Phillip J.R Goulder,4, Thumbi Ndung u University of KwaZulu Natal, HIV Pathogenesis Programme, Durban, South Africa, 2 Ragon Institute, Massachusetts General Hospital Harvard Medical School, Boston, MA, United States, 3 AIDS Research Institute IrsiCaixa, Hospital Universitari Germans Trias i Pujol, Barcelona, Spain, 4 University of Oxford, Department of Paediatrics, Oxford, United Kingdom Background: CD8 + T cell properties associated with virologic control of HIV- infection are not fully understood. In cross-sectional studies, T-cell factors associated with control include breadth of responses to Gag, T cell polyfunctionality and proliferative capacity. However, events responsible for the loss of control in some individuals remain unknown. We characterized longitudinally the immunological features that may explain divergence in disease outcome in 35 HIV- C-clade chronically infected individuals with protective alleles. Methods: Participants were enrolled in Durban, South Africa. Viral loads, CD4 counts and HLA typing performed by standard methods. HLAs A*74:0, B*57, B*58:0, B*8:0 were considered protective. HIV-specific CD8 + T-cell immune responses were quantified using the interferon gamma (IFN-g) enzyme-linked immunosorbent spot (ELISPOT) assay after stimulation with overlapping peptides (OLP) covering the whole HIV proteome. T cell polyfunctionality and proliferation upon stimulation with Gag peptide pool was assessed by flow cytometry and CFSE assay respectively. Results: At baseline, 5% were viremic controllers (VCs) (< 2,000 RNA copies/ml) and % were progressors (P) (> 00,000 RNA copies/ ml). Over time, 35% of VCs lost viral control. There was no significant difference in the overall magnitude or breadth of HIV- CD8 + T cell responses between P and VCs at baseline but VCs had higher breadth of Gag than P (p=0.0038). Among VCs that subsequently lost viral control, we noted a drop in the overall breadth of HIV- CD8 + T cell responses (p=0.0059). No significant differences were noted in T cell polyfunctionality or proliferation. Conclusions: Our data indicates that sustained HIV- control in C-clade infected patients with protective alleles is related to the breath of HIV- CD8 + T-cell responses against Gag. The loss of virologic control is related to a reduction in the total breath of CD8 + T cell responses in the absence of differences in polyfunctionality and proliferation. Paradigm-violating HLA Class II-restricted CD8 T-cells in HIV-infection Srinika Ranasinghe, Isaiah Davis, Damien Soghoian, Pedro Lamothe, John Sidney 2, Alessandro Sette 2, Mary Carrington, Hendrik Streeck 3, Louis Picker 4, Daniel Kaufmann, Bruce Walker Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 La Jolla Institute for Allergy and Immunology, La Jolla, CA, United States, 3 US Military HIV Research Program/WRAIR, Silver Spring, MD, United States, 4 Oregon Health & Science University, Portland, OR, United States Background: Classically, CD8 T cells recognize short peptides presented by a single HLA class I molecule, while CD4 T cells recognize longer peptides bound to HLA class II. However, Hansen et al Science 203 recently reported the induction of unconventional class II-restricted CD8 T cell responses in RhCMV/SIV strain 68- vector vaccinated macaques (reviewed in Ranasinghe & Walker, Nat Biotech 203). The detection of class II-restricted CD8 T cells in vaccinated macaques raises a critical question: do similar paradigm-violating class II-restricted CD8 T cell responses exist in natural HIV infection? Methods: We detected a class II-restricted CD8 T cell population in a treatment-naive HIV-infected individual. The response was identified using a novel CD8 HLA-DR ELISpot. In this assay, CD8 T cells exclusively targeted a single peptide, Gag4, presented by LCL stably expressing human recombinant HLA class II DRB* (in N=/22 DRB* subjects). Antibody blocking of HLA class I and II, and flow cytometric staining with a class II DRB*-Gag4 tetramer confirmed the class II CD8 restriction. Results: Strikingly, the DRB*-Gag4 tetramer specifically bound 2% of total CD8 T cells directly ex vivo (with no T cell expansion). The class II-restricted CD8αβ T cells exhibited a highly differentiated T EMRA memory phenotype. They also expressed high levels of Perforin and Granzyme B cytolytic proteins, and a distinct polyfunctional profile when compared intra-patient to the conventional class I-restricted B57-KF CD8 T EM response. Our data suggests the class II-restricted CD8 response is qualitatively and quantitatively distinct from conventional class I- CD8 T cells. Conclusions: In our exploratory study, we have demonstrated the first proof-of-principle detection of a large, unequivocal class II-restricted Gagspecific CD8 T cell response in an HIV-infected individual. Elucidating class II-restricted HIV-specific CD8 T cells that violate immunologic paradigms is likely to be important in future HIV vaccine design. ORAL ABSTRACT SESSIONS 23

124 Oral Abstract Sessions Oral Abstract Session 29: T Cell Immunity OA29.05 LB Anti-viral CD8 T-cells with B-cell Follicle Homing Potential Contribute to Vaccinemediated Enhanced Control of Pathogenic SIV Infection OA29.06 A Novel T-cell Vaccine Eliciting T-cell Specificities Associated with Control of HIV- In Humans Is Highly Immunogenic in Mice and Macaques ORAL ABSTRACT SESSIONS Geetha Mylvaganam, Daniel Rios 2, Ifor Williams 2, Vijayakumar Velu, Rama Amara Emory University, Yerkes National Primate Research Center, Atlanta, GA, United States, 2 Emory University, Department of Pathology, Atlanta, GA, United States Background: The germinal center (GC) resident T follicular helper cells (Tfh) represent a significant fraction of total pool of HIV/SIV infected cells during chronic infection and HAART. The GC are generally thought to exclude CD8 T cells and anti-viral CD8 T cells with potential to migrate to GC (follicular CD8) may enhance HIV/SIV control and reduce viral reservoirs. Here we studied the follicular CD8 in a cohort of DNA/ MVA vaccinated rhesus macaques (RM) that controlled or did not control a pathogenic SIV infection. Methods: RM were vaccinated with a DNA/MVA SIV vaccine and challenged intrarectally with SIVmac25. Animals with viral load below,000 copies at set point were defined as controllers. All controller RM (n=9) were vaccinated and non-controller RM (n=8) consisted of both vaccinated and unvaccinated. Results: Post challenge, we observed an aberrant enrichment of SIV+ PD- hi CD4 T cells in the LN and rectum of non-controllers but not controllers. The enhanced viral control was associated with higher frequency of Gag CM9 Tet+ CD8 T cells in the LN of controller RM compared to noncontroller RM. This was not evident in blood. Interestingly, a significant fraction of anti-viral CD8 T cells in the controller RM co-expressed CXCR5 (required for homing to B cell follicles/gc). The frequency of Tet+ CXCR5+ granzyme B+ cells was also higher in the LN of controller RM and higher frequencies correlated with lower Tfh and enhanced viral control. Immunofluorescence staining revealed co-localization of CD8 T cells with PD- bright cells in IgD- GC, a phenomena not observed in the non-controller RM. Impressively, the CXCR5+ CD8 T cells from the controller RM restricted the anti-cd3 driven expansion of CM9 peptide pulsed Tfh cells in vitro suggesting their killing potential. Conclusions: Our results reveal a novel subset of anti-viral CD8 T cells that may contribute to enhanced control of pathogenic SIV infection by infiltrating to GC of lymphoid sites and limiting SIV replication in Tfh in a vaccine setting. Beatriz Mothe,2, Xintao Hu 3, Anuska Llano, Margherita Rosati 3, Alex Olvera, Viraj Kulkarni 3, Antonio Valentin 3, Candido Alicea 3, Niranjan J. Sardesai 4, Muntsa Rocafort, Manel Crespo 5, Jorge Carrillo, Andrés Marco 6, James I. Mullins 7, Lucy Dorrell 8, Tomáš Hanke 9, Bonaventura Clotet,2,0, George N. Pavlakis 3, Barbara K. Felber 3, Christian Brander,2, IrsiCaixa AIDS Research Institute-HIVACAT, Hospital Germans Trias i Pujol, Badalona, Spain, 2 Universitat de Vic Universitat Central de Catalunya, Vic, Spain, 3 Human Retrovirus Section-National Cancer Institute, Frederick, MD, United States, 4 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States, 5 HIV Unit, Hospital de la Vall d Hebrón, Barcelona, Spain, 6 Centres Penitenciaris BCN, Barcelona, Spain, 7 University of Washington, Seattle, WA, United States, 8 MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, The John Radcliffe, Oxford, United Kingdom, 9 Jenner Institute, University of Oxford, Oxford, United Kingdom, 0 Universitat Autònoma de Barcelona, Barcelona, Spain, Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain Background: A top-down strategy was used to search for beneficial viral targets in large human immunogenicity data and to identify potential decoy targets that should be avoided in future vaccine designs. Methods: Through the identification of beneficial T cell responses in more than 000 individuals, the HIVACAT T-cell immunogen (HTI) was designed to contain 6 HIV- protein segments of 0-70 amino acids in length, covering >50 optimal defined CD4 + and CD8 + T-cell epitopes with >40 different HLA restrictions, without overrepresentation of B27/B57/ B58 restricted epitopes. Heterologous prime-boost regimens combining DNA (D) and MVA (M) expressing HTI were assessed in C57BL/6 mice and four Indian rhesus macaques. Cellular responses were characterized using IFN-γ ELISPOT and intracellular cytokine assays. Results: In C57BL/6 mice, DNA.HTI induced broad CD4 + and CD8 + T-cell responses to all segments within Gag, Pol, Vif and Nef. These responses were strongly increased by using heterologous regimens consisting of 3x DNA.HTI prime followed by MVA.HTI boost compared to 3 or 4x DNA.HTI only (median magnitude DDDM of 3,05 vs,40 and,353 SFC/0 6 splenocytes in DDD and DDDD groups respectively, p=0.0087). In rhesus macaques, DDD (administered IM in combination with macaque IL-2 DNA as molecular adjuvant using in vivo electroporation) induced % of specific T cells that persisted over a period of 4.5 months. MVA.HTI boosted the responses by 3- to 20-fold, reaching % IFN-γ T-cells. DDDM induced central and effector memory responses with a significant fraction of the vaccine induced IFN-γ + CD8 + T cell being either CD07a + or GzmB +. Conclusions: HTI delivered in a DDDM regimen was highly immunogenic in mice and macaques. The responses were CD8 + and CD4 + effector T cells with cytotoxic potential as well as CD4 + central memory T cells indicating long-term immunological persistence. These data justify further testing of the HTI approach in human clinical trials. 24 HIV Research for Prevention 204 HIV R4P

125 Oral Abstract Session 30: Antibody Functions and Protection Thursday, 30 October OA30.0 Synthetic Nucleic Acid Antibody Prophylaxis with Electroporation Drives Biologically Relevant Anti-HIV- Envelope Responses in vivo OA30.02 Insights on Synergistic Antibody-dependent Cellular Cytotoxicity (ADCC) Activity Mediated by Mutant Human Monoclonal Antibodies against HIV- Env Kar Muthumani, Seleeke Flingai, Megan Wise, Colleen Tingey, Kenneth E. Ugen 2, Niranjan Y. Sardesai 3, Joseph J. Kim 3, David B. Weiner University of Pennsylvania School of Medicine, Pathology and Lab. Medicine, Philadelphia, PA, United States, 2 University of South Florida Morsani College of Medicine, Department of Molecular Medicine, Tampa, FL, United States, 3 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States Background: Monoclonal Ab s have demonstrated therapeutic utility against several malignancies and infectious diseases. A drawback of this strategy is the time-consuming and expensive process requiring purification and scale up production of the Ab s for clinical use. A method to produce antibodies in vivo would be significant improvement for this platform. It would be important if these Ab s could be administered with out induction of vector serology allowing repeated administrations. Furthermore, delivery in a non-permanent fashion would also have advantages. Methods: Here we report development of new synthetic optimized plasmid vector/improved EP encoding Abs genes for delivery in vivo. This strategy allows for in vivo synthesis and serum expression of such ex vivo developed antibodies. Results: An enhanced and optimized DNA plasmid generates immunoglobulin heavy and light chains (Fab) of an established neutralizing anti-hiv monoclonal antibody (VRC0). We demonstrate that the serum of transfected animals exhibited the ability to bind to HIV envelopes in ELISA and FACS analysis against diverse isolates and this serum possessed HIV neutralizing activity equivalent to the native VRC0 antibody in vivo. In vivo delivery seroconverted the animals with in a few hours and neutralizing activity lasted for weeks. Antibodies exhibited a broad neutralization profile. This technology has important advantages for in vivo antibody production which could compliment or circumvent the need for standard antigen based vaccination, particularly in situations where there is difficulty in generation of protective antibody responses by immunization. Conclusions: This is the first study we are aware of using synthetic DNA plus EP delivery to produce circulating bioactive antibody responses in a living animal. The study has implications for prophylactic and therapeutic strategies for HIV and other important diseases especially in resource limited settings where antibody therapy is cost prohibitive. Chiara Orlandi, Anthony L. DeVico, Yongjun Guan, George K. Lewis University of Maryland, Institute of Human Virology, Baltimore, MD, United States Background: Several reports showed the key role of ADCC in protection against HIV- infection as it inversely correlates with progression in natural infection and with protection in non-human primate and human vaccines. Of note, optimal ADCC activity is the result of efficient antibody bridging between infected cells and FcγR-bearing effectors. We report that mab pairs attenuated for FcγR binding by mutagenesis mediate potent synergistic ADCC when admixed. Methods: We engineered the human mabs C, N5-i5 and N2-i2, specific for HIV- Env highly conserved epitopes, to exhibit three levels of anti-hiv ADCC strength: wild type, LALA variant and Fab fragment. ADCC and binding potency were assessed with Gp20 sensitized CEM NKr CCR5 target cells and the affinity to FcγRs by Elisa. We also developed a high throughput screening of ADCC-mediating antibodies combinations, quantifying the synergy with Chou-Talalay method. Results: Our ADCC assay revealed stronger activity for gp20 region C specific C and N5-i5 mabs then the Co-Rbs N2-i2 mab. Interestingly, LALA mutations entirely abrogated only N2-i2 cytotoxicity, while C and N5-i5 LALA retained weak ADCC activity at the highest concentrations tested. As expected, Fab fragments did not induce any target cells killing. Binding assays showed no difference in the affinity of all mabs variants for their viral antigens on target cells, meaning that the diverse cytotoxicity potency did not reside in the Fab regions. Moreover, the higher ADCC potency of C and N5-i5 wt and LALA more likely was due to different levels of binding to FcγR. Finally, we report for the first time that pairs of LALA mabs synergize strongly for ADCC with patterns unique to each mab pair. Conclusions: These data show that the antigen binding nature can overcome the loss of energy for IgG-FcγR interaction in LALA mutants. The findings provide a new experimental tool to clarify antigen-antibody aggregation contribution to Fc-mediated effector function and for passive immunization studies design. ORAL ABSTRACT SESSIONS 25

126 Oral Abstract Sessions Oral Abstract Session 30: Antibody Functions and Protection OA30.03 The Impact of Antiretroviral Treatment on HIV--Specific Broadly Neutralizing Antibody Responses OA30.04 Topical Application of Broadly Neutralizing Monoclonal Antibodies Reduces HIV Infection of Mucosal Tissue ORAL ABSTRACT SESSIONS Nonhlanhla N. Mkhize,2, Maphuti Madiga, Raveshni Durgiah, Elin S. Gray, Penny L. Moore,2,3, Sengeziwe Sibeko 3, Salim Abdool Karim 3, Lynn Morris,2,3, CAPRISA Acute Infection Study Team National Institute for Communicable Diseases of the NHLS, Virology, Johannesburg, South Africa, 2 University of the Witwatersrand, Johannesburg, South Africa, 3 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa Background: Initiation of antiretroviral therapy (ART) in HIV-infected individuals is associated with control of plasma viremia and improved CD4 counts which impacts on immune responses. We identified individuals from the CAPRISA 002 cohort with broadly cross-neutralizing (BCN) antibodies, who all initiated treatment within 4-6 years of infection. In this study we compared the neutralization titers and antibody specificties present before and after initiating ART. Methods: Total IgG was isolated (to remove traces of antiretroviral drug) using Protein G columns from plasma collected just prior to and within 2 years of ART from 6 women with BCN antibodies. These BCN antibodies were shown to target an N60-dependent epitope, a glycan at position 332 and in the MPER region on Env. Purified IgG was tested for neutralization against a panel of pseudoviruses (7 clade C, 4 clade B and 3 clade A) in TZM-bl cells. Neutralizing antibody specificities were mapped using pseudoviruses with mutations in key residues within each epitope. VSV-G was used as a control. Results: All plasma samples had IgG-associated neutralizing activity, with pre-art and post-art IgG fractions neutralizing the same viruses in the panel. However, there was a significant decline in neutralization titers in the post-art longitudinal samples. None of the IgG fractions neutralized VSV-G, indicating no residual ARV drug in these samples. Antibody epitope mapping data suggested that IgG antibodies isolated pre-art and after ART targeted the same epitopes. There was a significant decrease in viral loads and an increase in CD4 cell count as early as 6 months post-art initiation. Conclusions: Despite the low antigenic stimulation following ART, neutralizing antibodies were still present in most individuals with BCNs, even after 2 months of therapy. However, the decline in neutralization titers with preservation of antibody specificities during ART treatment suggests that the maintenance of robust BCN antibody responses is largely dependent on viral load. Yanille Scott, Kevin Whaley 2, Charlene S. Dezzutti 3,4 University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA, United States, 2 Mapp Biopharmaceutical, Inc., San Diego, CA, United States, 3 University of Pittsburgh School of Medicine, Pittsburgh, PA, United States, 4 Magee-Womens Research Institute, Pittsburgh, PA, United States Background: Broadly neutralizing monoclonal antibodies (nabs) reduce HIV transmission in vitro and in animal models of HIV transmission. However their efficacy in preventing human mucosal transmission of HIV remains unknown. During sexual transmission of HIV, semen is the inoculum and previous studies show that semen contains factors that modulate HIV infection in vitro. We theorize that topically applied nabs can reduce HIV infection of ectocervical and colonic mucosa in the context of semen. Methods: nabs, 4E0, VRC0, PG6 and PG9, were evaluated against HIV- JR-CSF ex vivo in polarized human ectocervical and colonic tissues. Tissues were treated with nabs and then inoculated apically with virus in the presence or absence of 50% whole human semen. Viral replication in tissues was monitored by HIVp24 ELISA for up to 2 days post inoculation. Data are presented as the median HIVp24 (pg/ml) and 95% confidence interval of 3-7 tissues from individual donors. ANOVA and post hoc multiple comparison procedures were used to compare nab and semen treatment groups. Results:.5µM nabs were required to protect ectocervical tissue compared to 0.03µM nabs in colonic tissue ex vivo. Treatment of ectocervical tissue with.5µm nabs resulted in protection of 90%, 00%, 80% or 30% of tissues by PG6, PG9, VRC0 and 4E0, respectively. Treatment with 0.03µM PG6, PG9 or VRC0, resulted in protection of 00% of colonic tissues, but only 50% with 4E0. Our combined results show that nab potency follows the order PG6>PG9>VRC0>>4E0, consistent between cell-based assays and ex vivo tissue studies. Similar levels of viral inhibition were observed in tissues treated with nabs in the presence of semen. Conclusions: Collectively, these data suggest nabs are effective in the presence of semen and should be considered as a non-chemotherapeutic option for the prevention of HIV. Importantly nabs could be considered for use in persons already infected with HIV and seeking ways to mitigate transmission to their partners. 26 HIV Research for Prevention 204 HIV R4P

127 Oral Abstract Session 30: Antibody Functions and Protection Thursday, 30 October OA30.05 OA30.06 LB Isolation of Monoclonal Antibodies from a SHIV-AD8 Infected Rhesus Macaque with Broad Neutralizing Activity Rebecca M. Lynch, Takuya Yamamoto, Patrick Wong, Ivelin Georgiev, Rodrigo Matus-Nicodemos, Stephen Schmidt, Rajeev Gautam 2, Zizhang Sheng 3, Lawrence Shapiro 3, Yoshiaki Nishimura 2, Malcolm A. Martin 2, John R. Mascola, Richard A. Koup Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 2 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 3 Columbia University, Department of Biochemistry and Molecular Biophysics, New York City, NY, United States Background: To assess the ability of immunogens to elicit broadly neutralizing antibodies (bnabs) against HIV-, an appropriate animal model is necessary. It has been previously established that the CCR5- tropic SHIV-AD8-EO molecular clone is a pathogenic and mucosally transmissible virus in Rhesus macaques. Furthermore, some macaques infected with this SHIV develop bnabs; however the mechanism of eliciting bnabs in this model remains unclear. Methods: One macaque, JG7, slowly developed breadth on a multiclade 2-virus panel from weeks 47-3 post-infection. Frequency of antigen-specific germinal center B cells and T-follicular helper (T FH ) cells were evaluated at the early wk47 time-point as well as somatic hypermutation in these B cell receptors (BCRs). Monoclonal antibodies were isolated and characterized by ELISA binding and neutralization profile. Results: By wk47, JG7 had high viral load, and high levels of antigenspecific T FH and IgG+ B cells in the germinal center. A fingerprint analysis of sera neutralization data suggested a CD4-induced (CD4i) specificity at the early wk47 time-point. BCR sequencing revealed a large clonal family distinguished by high V-gene mutation frequency and long heavy chain CDR3s of 3 amino acids. Four BCRs were cloned and found to recapitulate most of the serum breadth at this time. Furthermore, these monoclonals competed with 7b for binding to gp20 and had enhanced neutralization activity in the presence of soluble CD4, confirming the CD4i prediction. Conclusions: Our findings support the hypothesis that in the presence of high antigen levels and T cell help, Rhesus macaques can develop cross-neutralizing anti-hiv antibodies with high levels of mutation and long CDRH3s akin to bnabs isolated from HIV infected humans. These data highlight the utility of the SHIV model for studying development of bnabs in Rhesus macaques. Heterogeneous Conformation of HIV- Envelopes on Individual Virions Anush Arakelyan, Debbie King 2, Jean-Charled Grivel, Robin J. Shattock 2, Leonid Margolis NIH/NICHD, Section of Intercellular Interactions, Bethesda, MD, United States, 2 Imperial College, London, United Kingdom Background: HIV- has 7-5 envelope spike glycoproteins (Env) per virion consisting of a trimer of non-covalently linked gp20-gp4 heterodimers. These functional spikes are essential to virion binding and fusion, however it is unclear as to number required to mediate efficient fusion. Indeed other non-functional Env conformations, including uncleaved precursors (gp60), aberrant oligmers, monomers and gp4 stumps devoid of gp20 are also thought to be displayed on virions. The extent to which functional and non-function forms of Env are co-displayed on individual virions is currently a matter of debate. Recently, we developed a new technique, Flow Virometry that allows the study of proteins on the surface of individual virions. We have applied this technique to probe the conformation of Envs on the surface of individual particles using a panel of anti-env antibodies that discriminate between different conformations of these molecules. Methods: HIV- virions of BaL strain grown in PM CD4 T cells were captured with 5 nm magnetic nanoparticles (MNPs) coupled to one of several monoclonal antibodies recognizing particular conformations of Env. Captured virions were then stained with fluorescent anti-env antibodies different from the capture antibody and separated from free antibodies on a magnetic column. A range of neutralizing and nonneutralizing antibodies targeting functional and non-function Env forms were used to stain virions and their representation on individual viral particles assessed by flow virometry. Results: Our data indicate a non-uniform distribution of functional and non-functional Env within a viral population, where differential patterns of antibody staining revealed virion populations that were homogenous or mosaic with respect to functional and non-functional forms of Env. Conclusions: These finding have important implications for understanding antibody binding and neutralization, as well as other antibody effector functions. ORAL ABSTRACT SESSIONS 27

128 Poster Discussions Poster Discussion 0: Community Engagement and Advocacy PD0.0 Supporting Participant Adherence through Structured Engagement Activities in the MTN- 020 (ASPIRE) Trial Katie Schwartz, Patrick Ndase 2, Kristine Torjesen, Ashley Mayo, Rachel Scheckter, Ariane van der Straten 3, Lydia Soto-Torres 4, Thesla Palanee 5, Jared Baeten 2 FHI 360, Durham, NC, United States, 2 University of Washington, Seattle, WA, United States, 3 Women s Global Health Imperative; RTI International, San Francisco, CA, United States, 4 NIAID/DAIDS, Bethesda, MD, United States, 5 Wits Reproductive Health & HIV Institute, Johannesburg, South Africa Background: Sub-optimal product use in previous HIV prevention trials highlights the need for strategies to foster participant adherence to protocol requirements. Early in ASPIRE, participant engagement activities were implemented across all 5 study sites to enhance participant commitment within the prevention trial setting. Methods: Monthly tracking tools, site presentations and assessment visit summaries were reviewed to describe engagement activities, lessons learned and staff feedback. Proportion of ASPIRE participants engaged is reported from event attendance logs. Participant feedback is taken from staff debriefing reports of qualitative interviews. Results: Across ASPIRE sites, 76% of enrolled participants attended at least one of 99 staff-facilitated events held between 3/3-4/4. Events fell into 6 broad categories: milestone/retention events, social events, group education, male involvement events, couples workshops and holiday events. Support was fostered through peer educators, motivational speakers and shared experiences. Discussion topics included ring experiences, contraceptive options, adherence, impact of HIV, trial disclosure and community rumors. Social activities included movies, games and employment/life skills training. Lessons learned include designing events to be flexible and participant-centered, ensuring a mix of attendees, while maintaining participant confidentiality and comfort in group settings. Staff reported increased rapport with participants and improved team morale. Preliminary data from 28 participant interviews suggest that participants perceive engagement events as having a positive impact on personal commitment to trial and product use, as well as improved staff relations. Conclusions: Meaningful participant engagement is essential to HIV prevention trials with long follow-up and ongoing product use. Monitoring of participant engagement activities in ASPIRE will help inform design of future participant engagement and product support strategies in prevention trials. PD0.02 Pre-screening Strategies to Enhance Accrual and Retention in the ASPIRE Trial at the Wits Reproductive Health and HIV Institute, Johannesburg Pranitha Ramchuran, Krishnaveni Reddy, Patience Ramalo, Nombulelo Maseko, Amukelani Vuma, Lizzy Gama, Sylvia Sibeko, Nosipho Duba, Helen Rees, Thesla Palanee Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: The ASPIRE trial aims to assess the safety and effectiveness of Dapivirine Vaginal Ring for the prevention of HIV infection in women. In clinical trials, participant experiences at screening, visit waiting times and management of clinic flow impact accrual and retention significantly. To optimize these factors and improve operational and cost efficiencies in ASPIRE, novel pre-screening (PS) strategies were implemented. Methods: These strategies include recruitment of known HIV negative participants from Primary Health Care clinics within the catchment area, provision of voluntary counselling and testing on-site prior to the conduct of ASPIRE screening procedures and the implementation of a sitespecific PS checklist. The PS checklist lists basic ASPIRE eligibility criteria as well as documents/information required for screening (identification document, proof of contraception, details of 3 contacts). This tool is administered by community health workers (CHWs) during recruitment and allows determination of potentially eligible participants prior to their arrival at the clinic for screening procedures. It is also reviewed by the study administrators at the clinic when potential participants present for study screening in the event that information has changed. Results: The implementation of the above PS strategies results in the screening of only potential participants who meet basic ASPIRE eligibility criteria and reduces the number of participants who are screened out at a later stage. In addition, the PS Checklist reduces the frequency of split visits due to inadequate documentation/information by alerting participants to bring in all required information at their scheduled screening visit. Conclusions: These reductions in the number of unsuccessful/split screening visits reduce the clinic workload and participant waiting times. As a result, participant study experience is improved and study operational and cost efficiencies are optimized. This in turn impacts positively on accrual and retention. POSTER DISCUSSIONS 28 HIV Research for Prevention 204 HIV R4P

129 Poster Discussion 0: Community Engagement and Advocacy Wednesday, 29 October PD0.03 Why Should I Take Drugs for your Infection: Outcomes of Formative Research on Use of PrEP in Nigeria PD0.04 Consultation beyond the CAB: Engaging the Greater Community in Rectal Microbicide Clinical Trial Design and Planning John Idoko, Morenike O. Ukpong 2, Nancin Yusufu Dadem 3, Grace O. Kolawole 3, James Anenih 4, Emmanuel Alhassan 5 National Agency for the Control of AIDS, Abuja, Nigeria, 2 Institute of Public Health, New HIV Vaccine and Microbicide Advocacy Society, Ile-Ife, Nigeria, 3 Jos University Teaching Hospital, Department of AIDS Prevention Initiative Nigeria (APIN), Jos, Nigeria, 4 National Agency for the Control of AIDS, Strategic Knowledge Management, Jos, Nigeria, 5 National Agency for the Control of AIDS, Resource Mobilisation, Abuja, Nigeria Background: The study aimed to explore public opinion about PrEP including community interests and perceptions about the use of PrEP. Methods: The study used a mixed method approach. This included conduct of telephone interviews, in-depth interviews, focus group discussions, consultative meetings with critical stakeholders engaged in HIV prevention, treatment, care and support programs in Nigeria and an online survey to explore opinions about the design of the PrEP demonstration project in Nigeria. Information required include appropriate target group for the use of PrEP, community interest and perception about the use of PrEP as HIV prevention tool, how best to communicate about PrEP; concerns about PrEP use by sero-discordant couples; and suggestion for the design and implementation of the PrEP demonstration project Results: HIV sero-discordant couples were identified as the appropriate target group for the use of PrEP for a demonstration project in Nigeria. Most respondents felt PrEP would bring about added gains to the country by reducing HIV incidence and scaling-up of HIV prevention for key affected populations. Electronic and print media were identified as important means for massive public education about PrEP to prevent stigma and create awareness about PrEP. Stigma was identified as a major concern and a potential barrier for the uptake and use of PrEP by HIV sero-discordant couples. Most times, identified index partner is the sero-discordant relationship is a HIV positive female diagnosed during pregnancy. In a chauvinistic society like Nigeria, HIV negative male spouses may resist enrolling on a PrEP program because of the HIV positive status of the female partners. Conclusions: PrEP uptake and use by HIV sero-discordant couples in Nigeria may face challenges which are surmountable. A lot depends on appropriate actions taken by multiple players including motivation of HIV negative male partners to use PrEP, and effective public education programs to address stigma. Clare Collins, Cindra Feuer 2, Jerome T. Galea 3, Pedro Gonzales 4, Brian Kanyemba 5, Udom Likhitwonnawut 6, Jonathan Lucas 7, Steve Miralles 3, Benjamin Perkins 8, Jim Pickett 9, Pongpun Saokhieo 0, Wipas Wimonsate Microbicide Trials Network, Pittsburgh, PA, United States, 2 AVAC: Global Advocacy for HIV Prevention, New York, NY, United States, 3 Epicentro, Lima, Peru, 4 IMPACTA, Lima, Peru, 5 Desmond Tutu HIV Foundation, Cape Town, South Africa, 6 Consultant, Chiang Mai, Thailand, 7 Microbicide Trials Network, Durham, NC, United States, 8 Fenway Health, Boston, MA, United States, 9 International Rectal Microbicide Advocates, Chicago, IL, United States, 0 Research Institute for Health Sciences, Chiang Mai, Thailand, Thailand MOPH - U.S. CDC Collaboration, Bangkok, Thailand Background: Poor communication with key stakeholders and lack of community support and engagement can jeopardize HIV prevention research. This was demonstrated by earlier PrEP trials in Cambodia and Cameroon which were halted in response to pressure from advocates and activists. Indeed, these events might not have escalated to crisis levels had community and civil society representatives been invited to take part in the protocol development process or if they had been engaged before and during the course of these trials. Methods: Prior to the launch of the first ever Phase II rectal microbicide clinical trial (MTN-07), the Microbicide Trials Network (MTN) partnered with trial sites, some that were new to the MTN, and advocacy organizations to hold a series of civil society consultations. Six face-toface consultations were held during the study s protocol development stage in South Africa, Thailand, Peru and the U.S. Results: Consultation attendees (n= 8) gave feedback on an early draft of the MTN-07 study design and through in-depth discussions provided unique insight about regional and local social norms and practices, which resulted in significant changes to the study s design and implementation plan. For example, the protocol team heard from community stakeholders that the initial study design was confusing and that the eligibility criteria should include transgender women. Conclusions: The MTN-07 consultations moved beyond the traditional model focused on Community Advisory Boards (CABs) by securing feedback from individuals who were more closely representative of potential study participants and the community at large, benefitting the communities, trial sites and the MTN. This approach exemplifies the tenets of Good Participatory Practice Guidelines (GPP) for Biomedical HIV Prevention Trials jointly developed by AVAC and UNAIDS, which maintain effective communication and meaningful community engagement are essential for the successful and ethical conduct of HIV prevention trials. POSTER DISCUSSIONS 29

130 Poster Discussions Poster Discussion 0: Community Engagement and Advocacy PD0.05 The Impact of a Decade of Investment in HIV Prevention Research in Uganda Emmanuel Mugisha, Daniel Murokora 2, Bahati Prince Ngongo 3, Leslie Nielsen 4, Bonnie Bender 3 Independent Consultant, Country Director PATH, Kampala, Uganda, 2 Independent Consultant, Uganda Women s Health Initiative, Kampala, Uganda, 3 International AIDS Vaccine Initiative, Nairobi, Kenya, 4 International AIDS Vaccine Initiative, Kampala, Uganda Background: We reviewed the outcome of 0 years of investment in HIV new prevention technology (NPT) research including vaccine, microbicides and Pre-exposure prophylaxis (PrEP) in Uganda to identify areas of impact. Methods: In-depth interviews and focus group discussions were held with 9 key informants using the Research Impact Framework (London School of Hygiene and Tropical Medicine) to identify where HIV NPT investment has had the greatest impact. Secondary data was collected from four key research institutions in Uganda. Results: Investment in NPT research from has had a positive impact in Uganda. Respondents highlighted areas of high impact such as scientific contributions, including data demonstrating efficacy of Voluntary Medical Male Circumcision and PrEP; improved understanding of key populations epidemiology and increased scientific productivity as evidenced by increased publications. There have been significant gains in skills development and expansion of expertise; more than,000 clinicians, nurses, and counselors trained in Good Clinical Practice, specialized training of more than 800 lab scientists and technicians and sponsorship of more than 50 higher degrees. Improved clinical trial infrastructure and standards of community engagement were also highlighted. The investment impact was less than anticipated in the area of health system strengthening. Health benefits were limited to volunteer populations and to facilities in the vicinity of trial sites. In a few cases, improvements were not sustainable, expiring once projects were completed. Some respondents perceived international collaborations as being driven by external priorities resulting in upstream research and early product development being deprioritized. Conclusions: There have been clear benefits to HIV NPT research investment in Uganda. Future investments should consider how to incorporate objectives of local investigators, including upstream research, and ensure the sustainability of health system strengthening. POSTER DISCUSSIONS 30 HIV Research for Prevention 204 HIV R4P

131 Wednesday, 29 October Poster Discussion 02: Correlates of Protection in Highly Exposed Seronegative People PD02.0 Genital Proteome Correlates of Highly HIV- Exposed Uninfected African Women Romel D. Mackelprang, Kenzie Birsie 2, Mary J. Emond 3, Adam Burgener 4, Blake Ball 2, Jared Baeten 3, Connie Celum 3, Craig R. Cohen 5, Nelly R. Mugo 6, Jairam R. Lingappa 3, Partners PrEP Study Team University of Washington, Global Health, Seattle, WA, United States, 2 University of Manitoba, Winnipeg, MB, Canada, 3 University of Washington, Seattle, WA, United States, 4 University of Manitoba, Winnipeg, WA, United States, 5 University of California at San Francisco, San Francisco, CA, United States, 6 Kenya Medical Research Institute, Nairobi, Kenya Background: Identification of biologic factors robustly associated with resistance to HIV- infection among highly HIV exposed seronegative (HESN) individuals is a priority. We quantified HIV exposure among HIV- uninfected women in HIV serodiscordant couples and evaluated protein abundances as potential biomarkers of HIV resistance by comparing HESN versus low exposure (non-hesn) women. Methods: 33 vaginal swabs from 0 HESN (6 swabs) and 0 non- HESN (7 swabs) women in the placebo arm of the Partners PrEP Study had tandem mass spectrometry (MS) to assess frequency of 435 human and 247 bacterial proteins. We assessed differences by generalized estimating equations (GEE) adjusting for bacterial vaginosis, and calculated false discovery rates (FDRs). Results: Of HESN women, 00% reported unprotected sex versus 0% of non-hesn and had HIV infected partners with higher median plasma HIV RNA log 0 copies/ml (5.0 [interquartile range (IQR):4.9,5.]) than non-hesn women (.6 [IQR:.6,.9]). Based on these factors, HESN had >00-fold higher risk of infection than non-hesn. Hierarchical clustering of differentially abundant proteins distinguished HESN from non-hesn with 88% and 76% sensitivity and specificity, respectively. Nine proteins had FDR< 5%, including elafin, azurocidin and β2 integrin. Elafin was 8 times more abundant among HESN (95% CI=[ ];FDR=8.9x0-7 ), while β2 integrin was 4 times less abundant among HESN (95% CI=[.9-9.2];FDR=0.046).. Conclusions: The vaginal proteome differs in HESN and non-hesn women. Consistent with an earlier study, elafin, a mucosal anti-protease that may inhibit HIV, is more abundant among purportedly HIV resistant women; β2 integrin, which may facilitate cell-to-cell transfer of HIV, was decreased in HESN despite high exposure. Further research is needed to clarify if these factors mark HIV exposure or host resistance to HIV. PD02.02 Mx2 Expression Is Associated with Reduced Susceptibility to HIV Infection in Highly Exposed HIV Seronegative Sex Workers from Nairobi, Kenya Derek R. Stein, Souradet Y. Shaw 2, Max Abou 3, Stuart J. McCorrister 4, Garrett R. Westmacott 4, Francis A. Plummer 5, T. Blake Ball 3 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 2 University of Manitoba, Community Health Sciences, Winnipeg, MB, Canada, 3 Public Health Agency of Canada, National HIV and Retrovirology Laboratories, Winnipeg, MB, Canada, 4 Public Health Agency of Canada, Mass Spectrometry and Proteomics Core Facility, Winnipeg, MB, Canada, 5 Public Health Agency of Canada, Winnipeg, MB, Canada Background: Recent studies have identified Mx2 as a novel HIV- innate restriction factor that inhibits proviral integration. In these same studies Mx was found not to have an effect on HIV- in vitro. An initial proteomic study of immune cells from highly exposed HIV seronegative individuals (HESN) enrolled in the Pumwani sex worker cohort identified Mx and Mx2 as potential correlates of HIV protection. A detailed population level analysis of the role Mx and Mx2 contribute to reduced susceptibility to HIV infection was performed. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from 02 HESN women and 00 high-risk negative controls enrolled in a Nairobi-based sex worker cohort. Whole cell lysates were prepared and assayed for Mx and Mx2 expression by commercial ELISA. Bivariate and multiple linear regression analyses were conducted to control for age, pregnancy, menopause, contraception, recent infections, and medication use. Results: Mx2 was significantly overexpressed in HESN women compared to high-risk negative controls (p=0.027). No associations with Mx expression were observed. After multiple linear regression analyses accounting for age, menopause, pregnancy, Depo-Provera use, recent infections and medication usage, Mx2 expression was still significantly over-expressed in the PBMC of HESN women (p=0.04). Multivariate analysis also indicated that HESN women who use Depo-Provera have significantly higher levels of Mx2 than controls. Conclusions: This is the first epidemiological report of Mx2 and its association with altered susceptibility to HIV infection. These data corroborate findings from the in vitro HIV infection studies identifying Mx2 as a potent innate HIV restriction factor and suggests this molecule might have utility in HIV prevention strategies. POSTER DISCUSSIONS 3

132 Poster Discussions Poster Discussion 02: Correlates of Protection in Highly Exposed Seronegative People PD02.03 Low Immune Activation in the Ectocervix of HIV-exposed Seronegative Commercial Sex Workers Kristina Broliden, Anna Gibbs, Taha Hirbod, Terry B. Ball 2, Rupert Kaul 3, Joshua Kimani 4, Annelie Tjernlund Karolinska Institutet, Stockholm, Sweden, 2 University of Manitoba, Winnipeg, MB, Canada, 3 University of Toronto, Toronto, ON, Canada, 4 University of Nairobi, Nairobi, Kenya PD02.04 Plasma and PBMC mirna Profile in Sexually HIV- Exposed Seronegative Individuals Mara Biasin, Sara Yahyaei, Mariacristina De Luca, Irma Saulle, Federica Gnudi, Salomè Ibba, Micaela Garziano, Angela Berzi, Veronica Rainone, Daria Trabattoni, Sergio Lo Caputo 2, Francesco Mazzotta 2, Mario Clerici 3 University of Milan, Biomedical and Clinical Sciences, Milan, Italy, 2 S. Maria Annunziata Hospital, Florence, Italy, 3 Don Gnocchi Foundation, Milan, Italy Background: Heterogeneity in susceptibility to HIV infection has been demonstrated in HIV-exposed seronegative (HESN) individuals and resistance against infection may partly be attributed to a low mucosal immune activation state. Methods: Cervical tissue samples were collected from HESN female sex workers (FSW) and HIV seropositive FSW (HIV + FSW) from the Pumwani cohort of FSW, Nairobi, Kenya. HIV seronegative lower risk women (HIV - LR) from the same geographical region were also included as controls. Protein and mrna expression of selected immune markers were assessed by immunohistochemistry and qpcr, respectively, and compared between the study groups. The thickness of the ectocervical epithelium was analysed on sequential tissue sections. Results: The mrna levels of the cell makers CD3, CCR5, HLA-DR, DC-SIGN, Langerin, CD69 and of the pro-inflammatory cytokine markers IFN-alpha, IFN-gamma, TNF-alpha, IL-6, IL-7 and IL-22 were comparable between the HESN FSW and the HIV - LR study groups. Most of these markers were however significantly lower in the HESN FSW study group as compared to the HIV+FSW study group. Furthermore, the HESN FSW group had significantly lower mrna expression as well as protein expression of CD4 + cells as compared to the other two groups and they had significantly more mrna and protein expression of CD8 as compared to the HIV - LR group. The ectocervical epithelium of the HESN FSW samples was generally thicker than that of the other two study groups. Conclusions: Despite active commercial sex work these relatively resistant HESN FSW had a low immune activation state as assessed in ectocervical tissue samples. They also had relatively low numbers of HIV target cells (CD4+ cells) and a thicker epithelium than the control groups. This mucosal immune phenotype may be beneficial in the resistance against HIV infection. Background: MicroRNAs (mirnas) are small 20- to 24-nt non-coding RNAs involved in the post-transcriptional regulation of gene expression which play important defensive roles in several viral infections. Global expression profiles of cellular mirnas have identified alterations of specific mirnas post-hiv- infection both in vitro and in different patient cohorts suggesting potential roles for mirna in pathogenesis and disease progression. We therefore decided to verify if natural resistance to HIV- infection observed in seronegative individuals repeatedly exposed to HIV- (HESN) through unprotected sexual intercourse could be secondary to a different expression of their mirna profile. Methods: expression levels of 25 mirnas selected according to their proven anti-hiv- properties were analyzed in plasma, basal PBMC and in in vitro HIV- infected macrophages isolated from 30 HESN, 30 HIV seropositive subjects (HIV+) and 30 healthy controls (HC). Results: In plasma the expression of mir-55, mir-382, mir-28 and mir- 98 was significantly augmented in both HIV+ and HESN compared to HC probably as a consequence of viral exposure. Conversely the expression of mir-223 and mir-50 in plasma was significantly increased only in HESN and this result was also confirmed in basal PBMC suggesting a protective effect for these mirnas in resistance to HIV- infection. Furthermore, the expression of mir-50 was significantly increased in HESN macrophages following HIV- infection. Conclusions: mir-223 and mir-50 can target the 3 UTR of HIV- transcripts, and they have already been identified as anti-hiv- mirnas. The higher expression of these mirna in HESN samples could therefore represents a key protection mechanism against HIV infection. POSTER DISCUSSIONS 32 HIV Research for Prevention 204 HIV R4P

133 Wednesday, 29 October Poster Discussion 02: Correlates of Protection in Highly Exposed Seronegative People PD02.05 The Role of Hormones in Natural Protection against HIV- in the Kenyan HIV-exposed Seronegative Cohort Aida Sivro, Ruey-Chyi Su, Max Abou 2, Joshua Kimani,3, Walter Jaoko 3, Adam Burgener, Frank Plummer,2, T. Blake Ball,2,3 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 2 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada, 3 University of Nairobi, Medical Microbiology, Nairobi, Kenya Background: Studies of populations at high risk of HIV- infection have identified subsets of individuals who are HIV-exposed but remain seronegative (HESN). Work is in progress to understand the mechanisms contributing to the observed natural protection against HIV-infection. Sex hormones, which play crucial roles in regulating immune function, are also shown to contribute to increased prevalence of sexually transmitted infections in women. Hormonal contraceptive use and pregnancy, a state of high estrogen and progesterone levels, have been associated with increased risk of HIV- acquisition. Our earlier study showed association of three polymorphisms in interferon regulatory factor (IRF) gene with HIV-resistant phenotype and changes in IRF regulation. This study examined the plasma estrogen, progesterone, cortisol and prolactin hormone levels with respect to IRF polymorphisms and HESN phenotype. Methods: Samples used for this study were obtained from the Majengo Sex Worker Cohort in Nairobi, Kenya. Plasma hormone levels were measured using Millipore Milliplex Hormone Magnetic Bead kits (Human Pituitary and Steroid/Thyroid Panels) and analyzed on the BioPlex-200. Results: Significantly lower prolactin levels were observed in the plasma samples from women with protective IRF genotypes when corrected for age, use of hormonal contraceptives and menstrual cycle. Interestingly, levels of all four hormones examined were significantly lower in the plasma samples from HESN women (with or without the protective IRF- genotypes) compared to HIV-Susceptible women from the same sex worker cohort. Conclusions: These results indicate that hormonal regulation of the immune environment could be one of the key contributing factors to the natural resistance against HIV-infection observed in the Kenyan Majengo cohort. Understanding how hormone levels manipulate the microenvironment of the female genital tract could greatly contribute to the development of improved immunization strategies against HIV-. POSTER DISCUSSIONS 33

134 Poster Discussions Poster Discussion 03: Preclinical and Clinical Vaccine Trials PD03.0 LB CD8 + T-cell Mediated HIV Inhibition after Vaccination with a DNA/Recombinant Ad5 (rad5) HIV Vaccine Is Similar to that Seen in Treated HIV Infection PD03.02 DNA/MVA and Protein-based SIV Vaccine Regimens Delivered to Rhesus Macaques with Novel Adjuvants Fail to Elicit Neutralizing Antibodies with Breadth POSTER DISCUSSIONS Nicole Frahm,2, Bryce A. Manso, Stephen C. De Rosa,3, Christina Ochsenbauer 4, Shelly Karuna, Magdalena Sobieszczyk 5, Scott Hammer 5, Edith Swann 6, Barney Graham 7, Peter Gilbert,8, Margaret Juliana McElrath,2,3, the NIAID HIV Vaccine Trials Network Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 University of Washington, Department of Global Health, Seattle, WA, United States, 3 University of Washington, Department of Laboratory Medicine, Seattle, WA, United States, 4 University of Alabama at Birmingham, Birmingham, AL, United States, 5 Columbia University, New York, NY, United States, 6 Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 7 Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 8 University of Washington, Department of Biostatistics, Seattle, WA, United States Background: HVTN 505, a phase 2b trial assessing if a DNA/rAd5 HIV vaccine prime/boost regimen reduces HIV acquisition or viremia post infection, halted vaccinations in 203 due to efficacy futility, though it induced substantial CD8 + T-cell responses. We examined whether vaccine-induced T cells in HVTN 505 were unable to inhibit virus replication, as a potential reason for their inability to control viremia post infection. Methods: CD8 + T cells from 48 participants (38 vaccine, 0 placebo) were tested pre-immunization and at peak immunogenicity (4 weeks post rad5 boost) for their ability to inhibit HIV- replication in autologous CD4 + T cells infected with a NanoLuc -secreting JR-CSF infectious molecular clone. Relative light units (RLU) were measured at day 7, Δlog inhibition of virus (difference of log RLU in the presence vs. absence of CD8 + T cells) is reported for an effector:target ratio of 5:. Responses were considered positive if above the mean + 3SD of baseline and placebo samples (.05 log). Seven subjects with chronic HIV infection on antiretroviral treatment (ART) served as controls. Results: Viral inhibition by CD8 + T cells was significantly greater in vaccinees at peak immunogenicity than pre-immunization (p< 0.000); no changes over time were observed in placebo recipients (p=0.7). The response rate was 6%, with median Δlog inhibition of.9 logs (range.2-2.6) in responders. This level of inhibition is similar to that in HIVinfected subjects requiring ART to control viremia (median 2. logs, range.7-2.8; p=0.06). Conclusions: The DNA/rAd5 vaccine in HVTN 505 led to significant induction of CD8 + T-cell responses able to inhibit HIV- replication in vitro, but the magnitude in responders was comparable to that observed in treated HIV infected subjects requiring ART to control viremia. It is thus likely that vaccination must elicit responses of higher magnitude (such as those observed in elite HIV controllers in similar assays) to drive an overall reduction in viral load post infection. Megan K. Murphy, Katherine S. Wetzel 2, Katie M. Kilgore, Stacey A. Smith, Samantha L. Burton, Sharmila Reddy, Nicholas Francella 2, Donald L. Sodora 3, Guido Silvestri, Kelly S. Cole 4, Francois Villinger, James E. Robinson 5, Bali Pulendran, Ronald G. Collman 2, Rama R. Amara, Cynthia A. Derdeyn Emory University, Atlanta, GA, United States, 2 University of Pennsylvania, Philadelphia, PA, United States, 3 Seattle Biomedical Research Institute, Seattle, WA, United States, 4 University of Pittsburgh, Pittsburgh, PA, United States, 5 Tulane University, New Orleans, LA, United States Background: Antibodies that neutralize diverse HIV- strains are likely to be important for a protective vaccine. To this end, pre-clinical SIVbased nonhuman primate immunization regimens have been designed to enhance antibody-mediated protection. Here we investigated whether neutralizing antibody breadth was elicited by four different DNA/MVA and protein-based vaccination regimens that included novel adjuvants such as GM-CSF and TLR ligands, but expressed the same SIVmac239 envelope (Env) glycoprotein. Methods: A novel panel of 4 diverse SIV Envs described in previous studies was generated. Neutralization and inhibition assays were performed using Env pseudoviruses in the Tzm-bl assay. CD4 independent entry was evaluated using 293T cells expressing rhesus CCR5 with or without CD4. Results: SIV-infected plasma and monoclonal antibodies targeting V3, CD4-induced structures, and the CD4 binding site neutralized four of the Envs, designated as tier. Three tier Envs were also CD4 independent, and only one carried a recently described signature of neutralization sensitivity (45T/47R) in the gp20 C region. The remaining 0 SIV Envs had varying levels of neutralization resistance. We tested sera from 9 immunized rhesus macaques for neutralization breadth against the SIV Env panel. The pooled vaccinated monkey sera from each of the four trials neutralized only the sensitive tier Envs, regardless of immunization regimen, and there was no difference between serum from animals that became infected or remained protected after 2 mucosal challenges within each trial. Conclusions: These results indicate the SIVmac239 Env immunogen was the primary determinant of the neutralizing antibody profile, suggesting that elicitation of antibodies with greater neutralization breadth will likely require modification or careful selection of the Env immunogen itself. In addition, this panel of SIV Envs provides a useful tool for evaluating the neutralization capacity and breadth of antibodies elicited by SIV-based vaccines. 34 HIV Research for Prevention 204 HIV R4P

135 Poster Discussion 03: Preclinical and Clinical Vaccine Trials Wednesday, 29 October PD03.03 Antigenicity and Immunogenicity of Disulfidestabilized HIV- Envelope Glycoprotein Oligomers PD03.04 LB Clinical Safety and Immunogenicity of Two HIV Vaccines SeV-G (NP) and Ad35-GRIN in HIV-uninfected, Healthy Adult Volunteers Yu Feng, Hao Jiang, Jidnyasa Ingale 2, Krisa McKee 3, John Mascola 3, Richard Wyatt,2 International AIDS Vaccine Initiative, IAVI Neutralizing Antibody Center at TSRI, La Jolla, CA, United States, 2 The Scripps Research Institute, La Jolla, CA, United States, 3 National Institute of Allergy and Infectious Diseases, Vaccine Research Center, Bethesda, MD, United States Background: Our previous study suggests that YU2 gp20 trimers, deleted of their major variable regions and partially stabilized in the CD4-bound state display better elicitation of CD4 binding site (CD4bs)- directed neutralizing antibodies compared to core and full-length trimers. The elicited neutralizing antibodies target elements exposed only on Env of sensitive viruses and cannot neutralize more resistant isolates. Structural studies reveal a layered gp20 architecture in which three topologically separate and structurally plastic layers act as a shapechanging spacer, facilitating movement between the outer domain and gp4 to allow movement among alternative conformations required for virus entry and immune evasion. A recent report demonstrates that disulfide bond (C65-C5) inserted between layers can lock gp20 in a CD4 bound state. Methods: Here, we integrated the disulfide bonds C09-C428 (inner and outer domain), C65-C5 (layers and 2) and C95-C484 (layers 2 and 3), into the YU2 gp20 trimers. Results: These stabilized Envs displayed improved stability and antigenic profiles. The substituted cysteines eliminated binding of most non-neutralizing CD4bs antibodies while retaining recognition by most broadly neutralizing CD4bs antibodies. Armed with this promising antigenic profile, we immunized guinea pigs and non-human primates with these disulfide-stabilized Envs with or without conjugation onto liposome particles. The parental trimers once again elicited Tier neutralization, however, introduction of the C09-C428 rendered antibodies unable to neutralize Tier viruses. However, the stabilized Envs elicited improved binding antibodies toward the CD4bs and, in addition, conjugation onto liposomes shifted the immune responses to the CD4bs. Conclusions: These results suggest that structure-guided stabilization can be used to better expose conserved epitopes of gp20 and the immune responses can be shifted onto the conserved epitopes by particulate display or by other stabilization strategies. Etienne Karita, Omu Anzala 2, Brian Gazzard 3, Philip Bergin 4, Julien Nyombayire, Gloria Omosa 5, Akil Jackson 6, Rosine Ingabire, Gina Ouattara 5, Harriet Park 7, Anne Gumbe 5, Kundai Chinyenze 5, Sabrina Welsh 7, Carl Verlinde 7, Lorna Clark 4, Paramesh Chetty 8, Mumtaz Booley 8, Jean Bizimana, Bashir Farah 5, Peter Hayes 4, Devika Zachariah 7, Kristen Syvertsen 7, Michele Fong Lim 7, Len Dally 9, Burc Barin 9, Makoto Inoue 0, Hiroto Hara 0, Takashi Hironaka 0, Tsugumine Shu 0, Mamoru Hasegawa 0, Tetsuro Matano, Eddy Sayeed 7, Christopher Parks 7, Jim Ackland 2, Patricia M. Fast 7, Jill Gilmour 4, Josephine H. Cox 7, Angela Lombardo 7, Dagna Laufer 7 Project San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 2 Kenya AIDS Vaccine Initiative-Kangemi, University of Nairobi, Nairobi, Kenya, 3 Chelsea and Westminster Healthcare National Health Service Foundation Trust, London, United Kingdom, 4 International AIDS Vaccine Initiative-Human Immunology Lab, Imperial College, London, United Kingdom, 5 Kenya AIDS Vaccine Initiative University of Nairobi, Nairobi, Kenya, 6 Chelsea and Westminster Healthcare National Health Service Foundation Trus, London, United Kingdom, 7 International AIDS Vaccine Initiative, New York, NY, United States, 8 International AIDS Vaccine Initiative, Johannesburg, South Africa, 9 EMMES Corporation, Rockville, MD, United States, 0 DNAVEC Corporation, Tsukuba, Japan, University of Tokyo, Tokyo, Japan, 2 Global BioSolutions, Melbourne, Australia Background: Development of vaccines that stimulate sustained humoral and/or cellular immunity at mucosal HIV entry points is critical in the quest for an HIV vaccine. To achieve this goal, we are investigating replication-competent viral vectors for mucosal delivery that might mimic the efficacy of live-attenuated viral vaccines. Methods: A prototype HIV vaccine based on replication-competent Sendai virus expressing subtype A Gag [SeV-G(NP)] was manufactured. SeV-G(NP) was administered intranasally (IN) in heterologous prime boost (P/B) combinations with an Adenovirus-35 encoding subtype A Gag, RT, Integrase and Nef (Ad35-GRIN at x0^0 vp) given intramuscularly (IM) or in a homologous regimen, all at 0 and 4 months: Groups A and B were dose escalations (2x0^7, 2x0^8 CIU SeV-G(NP), respectively) followed by Ad35-GRIN, Group C: Ad35-GRIN followed by 2x0^8 CIU SeV-G(NP); Group D: two doses of 2x0^8 CIU SeV-G(NP). Sixty-five HIV uninfected adults were enrolled in Kenya, Rwanda and the United Kingdom. Systemic and mucosal cellular and humoral immune responses were assessed. We present here preliminary Interferongamma (IFN-y) ELISPOT and serum Gag-p24 binding antibody data. Results: All vaccine regimens were well-tolerated with no vaccine-related SAEs. Systemic HIV-specific IFN-y ELISPOT responses were seen in all recipients but of the heterologous P/B regimen of SeV-G(NP) followed by Ad35-GRIN. In Groups A and B the magnitude and response rate of Gag specific T-cell responses indicated that the SeV-G(NP) provided a strong priming effect ( hidden prime ), superior to that seen with other primes such as DNA. Systemic IgG and IgA Gag-p24 antibody responses were detected in recipients of the heterologous P/B regimen of Ad35- GRIN followed by SeV-G(NP). Conclusions: The combination of IN SeV-G(NP) and IM Ad35-GRIN was well tolerated. The SeV-G(NP) vaccine appears to enhance both cellular and antibody responses to Ad35-GRIN. The order of vaccination appears to determine which immunogenic response is stimulated. POSTER DISCUSSIONS 35

136 Poster Discussions Poster Discussion 03: Preclinical and Clinical Vaccine Trials PD03.05 LB Learning from the Success of HPV Vaccines to Develop HIV Vaccines that Break B Cell Selftolerance John T. Schiller, Bryce Chackerian 2 National Cancer Institute, Laboratory of Cellular Oncology, Bethesda, MD, United States, 2 University of New Mexico School of Medicine, Department of Molecular Genetics and Microbiology, Albuquerque, NM, United States Background: Even a single dose of an HPV vaccine composed of L virus-like particles (VLPs) consistently induces high titer and durable neutralizing antibody (nab) responses that afford high vaccine efficacy against incident HPV infection. Methods: We believe that the exceptionally potent response induced by this non-infectious subunit vaccine is largely due to the dense repetitive display of epitopes on the surface of the VLPs. Results: Correspondingly, we found that ordered repetitive display of central self-antigens on VLPs, at a spacing of less than 50Å, efficiently breaks self-tolerance, resulting in high titer auto-antibodies. Self-antigen display with this spacing can even reactivate anergic B cells induced by self-tolerance mechanisms. It follows that HPV VLPs, which have typical virus epitope spacing of 50-00Å, should be able to induce nab through immunoglobulin gene developmental pathways that involve self-reactive intermediates. Since the majority of naïve B cells are self-reactive, this ability likely makes a critical contribution to the consistently potent nab responses after even a single VLP dose. Conclusions: HIV has an exceptionally low number of receptor binding determinants on its surface. The paucity of envelope spikes likely contributes to its exceptionally low transmission rate. The spacing of HIV envelope spikes has been estimated to be 230Å, and so we postulate that neutralizing antibodies to HIV, unlike typical viruses, cannot develop through self-reactive intermediates. This might explain why nab, especially broadly cross-neutralizing ones, are produced through highly convoluted developmental pathways. In sacrificing transmission efficiency for delayed induction of nab, HIV may have evolved an Achilles heal that could be exploited by vaccines that can generate nabs through self-reactive intermediates. We believe that development of vaccines displaying HIV envelope epitopes at high density should be a major focus HIV vaccine development. POSTER DISCUSSIONS 36 HIV Research for Prevention 204 HIV R4P

137 Poster Discussion 04: Behavioral and Social Sciences Thursday, 30 October PD04.0 PD04.02 VOICE-C Participant Narratives of Rape: What they Mean for Female-initiated HIV Prevention Products Miriam A. Hartmann, Elizabeth T. Montgomery, Jonathan Stadler 2, Nicole Laborde, Ariane van der Straten,3, on behalf of the VOICE-C team RTI International, Women s Global Health Imperative, San Francisco, CA, United States, 2 University of the Witwatersrand, Wits Reproductive Health and HIV Institute, Faculty of Health Sciences, Johannesburg, South Africa, 3 UCSF, Center for AIDS Prevention Studies, Department of Medicine, San Francisco, CA, United States Background: Numerous recent trials have sought to develop femaleinitiated methods of HIV prevention; in part to address barriers to women s prevention rooted in unequal gender norms. Understanding the gender-related context in which these products may be used, including levels of violence against women, is critical. MTN-003C (VOICE-C), a qualitative sub-study of the MTN-003 (VOICE) HIV prevention trial, examined socio-cultural barriers and facilitators to product use within the Johannesburg site. Methods: We conducted and analyzed focus group discussion (FGD), in-depth interview (IDI), and serial ethnographic interview data from 02 female VOICE participants, 22 male partners, 7 community advisory board (CAB) members, and 23 community stakeholders. Violence was not a designated interview topic; however it arose through discussions of community context, interpersonal relationships, and product use. For this analysis, all textual data coded as violence was systematically reviewed. Results: The data revealed the prominence of sexual violence in women s lives. Rape was discussed in 2/4 FGDs with CAB members, 2/3 FGDs with stakeholders and among a quarter of interviews/fgds with VOICE female participants; two of whom described personal experiences of rape. These narratives demonstrated a pervasive perception that women are vulnerable to rape and that this threat contributes to their susceptibility to HIV. The possibility of rape was used to reframe their HIV risk as external to their own or their partner s behavior and was ultimately used to rationalize the importance of female-initiated products in HIV prevention. Conclusions: Fear or experience of rape is pervasive in this community, reflecting underlying gender inequalities, which in turn are likely to influence how HIV prevention products are perceived and used. While the actual impact on product use in VOICE is uncertain, results illustrate how women, in contexts of high sexual violence, may utilize existing unequal gender norms to negotiate their use. He Said, She Said. Exploring Couples Sensory Perceptions and Experiences with Vaginal Gels & Film: Implications for Microbicide Development Kathleen M. Morrow, Rochelle K. Rosen 2, Joseph L. Fava 3, Lisa Rohan 4, Erna M. Kojic 5, David Friend 6, David Katz 7, Robert Buckheit 8 Miriam Hospital & Alpert Medicial School of Brown University, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 2 Miriam Hospital & Brown University School of Public Health, Center for Behavioral and Preventive Medicine, Providence, RI, United States, 3 Miriam Hospital, Center for Behavioral and Preventive Medicine, Providence, RI, United States, 4 Magee Womens Research Institute, Pittsburg, PA, United States, 5 Miriam Hospital & Alpert Medical School of Brown University, Division of Immunology, Providence, RI, United States, 6 CONRAD, Arlington, VA, United States, 7 Duke University, Biomedical Engineering and Ob/Gyn, Durham, NC, United States, 8 ImQuest BioSciences, Inc., Frederick, MD, United States Background: Perceptibility science, the objective measurement of user sensory perceptions (sensations) and experiences (USPE) of formulation performance during use, is a new concept. For vaginal gels, sets of rheological and other biophysical properties, including measures of spreading and retention, may critically impact user experiences. For vaginal films, other properties, e.g., disintegration time or puncture strength, may play additional roles. Methods: In a mixed methods study, 24 monogamous HIV-/STIheterosexual couples completed 3 formulation evaluation visits (00% retention). Following vaginal sex, participants (ppts) independently rated USPEs for 2 volumes of HEC gel (2 ml; 4 ml) and x2 vaginal film. 8 USPE scales were scored at each visit for each participant. Pairwise comparisons between females (F) and males (M) were conducted on each form. Results: There were no significant differences between F and M USPE evaluations for low volume gel. For high volume gel, there was a significant difference in Initial Penetration scores, with F>M (4.3 v 3.8, p=.02; large effect size (ES) d=.78). Perceived Leakage scores were also higher in F than M (2.4 v 2.; medium ES d=.53), and near significant (p=.08). For film, a significant difference was seen in the Stimulating scale score with M>F (2.0 v.4, p=.04; large ES d=.77). All other USPE scale score comparisons (2 of 24) across F & M were strikingly similar. Both F & M ppts reported greater sensory experiences overall with gels than films. High volume gel was slightly favored in both genders (F=, M=0). Contrasts occurred in choices between low volume gel and film: more F chose low volume gel (F=9) and more M chose film (M=9). Conclusions: A non-optimized user experience could compromise an optimized drug and its delivery. Perceptibility, for both members of a sexual dyad, should be considered when designing and advancing potential vaginal (and rectal) formulations for HIV/STI prevention and multipurpose prevention technologies. POSTER DISCUSSIONS 37

138 Poster Discussions Poster Discussion 04: Behavioral and Social Sciences PD04.03 PD04.04 POSTER DISCUSSIONS Successfully Addressing Challenges to Implementing a Multinational SMS-based Reminder and Data Collection System in a Biomedical HIV Prevention Trial William Brown III,2, Rebecca Giguere, Mobolaji Ibitoye, Alex Carballo-Diéguez, Ross D. Cranston 3 Columbia University, HIV Center for Clinical and Behavioral Studies, New York, NY, United States, 2 Columbia University, Biomedical Informatics, New York, NY, United States, 3 University of Pittsburgh, Department of Medicine, Pittsburgh, PA, United States Background: Adherence to product use in a clinical trial is critical to assessing safety and efficacy. Real-time data collection can help monitor adherence. The ubiquity of Short Message Service (SMS) allows researchers to collect adherence data and provide behavioral reminders remotely in real-time. MTN-07 is a phase 2 safety and acceptability study of oral Truvada tablets and rectally-applied tenofovir gel. To monitor adherence in MTN-07, we sought to ) identify an SMS system that allowed for daily reminders and collection of data on product use, 2) implement it in four countries and five languages, and 3) develop a centralized data management system with an automated backup system. Methods: We assessed feature availability of several SMS systems based on ideal criteria, including keyword response, capability, participant identification and grouping, text message scheduling, multiple language operating system, and international SMS capability. After identifying the optimal SMS system, we systematically implemented it in each country, working with IT staff at clinical research sites. Results: We successfully addressed 24 critical challenges pre- and post-implementation. Solutions included: developing a federated SMS-system architecture to mitigate SMS message costs and manage data access; using secure protocols to centralize data backup, developing several programming syntaxes to facilitate daily data analysis, developing a calendar template for reporting SMS behavior to sites, ambiguation of text message language to increase privacy, and standardizing operating systems and hardware to minimize variability in system performance. Other solutions and metrics for estimating cost effectiveness will be discussed. Conclusions: Our tests and continued use have allowed us to identify factors that should be consistent across countries to ensure smooth implementation and operation of SMS as an adherence reminder system and real-time data collection modality. Reimagining HIV Testing in an Era of ART Martina Brostrom, Ruben Granich, Somya Gupta, Badara Samb UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva, Switzerland Background: UNAIDS launched Treatment 205 to accelerate treatment access through the concepts of innovation, speed and focus. In 203,2,9 million of 35 million people living with HIV globally were receiving treatment. UNAIDS has proposed a target for % of people living with HIV tested, 90% of those tested on ART and 90% of those on ART virally suppressed. Ensuring treatment and viral suppression for people living with HIV will require special efforts to ensure that people living with HIV but who do not know their statushave access to HIV testing. Methods: We reviewed the national strategies of 0 high impact countries in Africa and identified gaps and strategic opportunities for innovating HIV testing. Results: Harnessing the therapeutic and preventive benefits of ART, will entail a massive scale up and reorientation of testing efforts in countries where HIV testing deficits limit access to ART. An estimated 55% of people living with HIV in SSA are unaware of their HIV status. Data from select countries of people ever tested in SSA, where treatment coverage is 37%, ranges from very low to moderate or high and with some countries having a significant gender gap (where the proportion of men ever tested is low but the range among women is moderate or high). Often testing coverage is mismatched with the epidemic realities of countries and testing policy is not in aligned with treatment expansion targets. This calls for revised programmatic targets for testing and treatment, high yield strategies and new benchmarks for quality HIV care, with an attention to location, gender and age. Conclusions: HIV testing programmes in many countries are inadequatethey were conceptualized when AIDS was an emergent and frightening phenomenon- now they need to address mature epidemics, where there is approaching universal access to treatment which allows HIV infection to be non-life threatening, and where the concentrations of HIV in the most affected populations in different contexts is much more known. Key areas for innovation include self-testing, multi-disease community campaigns and PoC testing for CD4 and VL for first line service delivery. 38 HIV Research for Prevention 204 HIV R4P

139 Poster Discussion 04: Behavioral and Social Sciences PD04.05 Thursday, 30 October The Need for Demonstration Projects to Ensure Key Populations Gain Access to New HIV Prevention Biomedical Tools in South Africa Brian Kanyemba, Ben Brown, Maaza Seyoum 2 Desmond Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town, South Africa, 2 International AIDS Vaccine Initiative (IAVI), Johannesburg, South Africa POSTER DISCUSSIONS Background: Sero-discordant couples and Key Populations(KP) - including, but not limited to, men who have sex with men (MSM), sex workers (SWs) and people who inject drugs (PWID) - urgently need new, effective HIV prevention methods. Numerous studies have shown that pre-exposure prophylaxis (PrEP) and treatment as prevention (TasP) offer effective HIV prevention to these vulnerable groups. Prior to the 203 South African AIDS Conference, a pre-conference satellite was conducted to gain key stakeholders perspectives on the implementation of demonstrations projects for KP. Methods: After a plenary session that reviewed current HIV prevention research and advocacy, 50 delegates were divided into four groups to debate opportunities and challenges for demonstration projects. The two key questions were: ) Which HIV prevention research methods should be prioritized for demonstration projects among KP in SA; and 2) What will the main challenges be? The session was divided into four 5-minute focus groups, where the delegates circulated through topics. Facilitators captured key findings. Results: There was strong agreement that PrEP and TasP should be available to MSM, SW, and PWID in SA. In order to achieve this goal, demonstration and pilot projects would be a valuable source of data on the implementation of ARV based prevention outside the clinical trial setting. The key recommendations focused on the need to implement demonstration projects that are tailored to priorities, needs and circumstances of diverse KP groups. Such projects should answer questions on adherence, the impact of stigma in health care settings, and access to care despite structural barriers such as criminalization of some KP. Conclusions: Stakeholders who participated in the session highlighted need for PrEP access among KP in SA (and other African countries) in order to begin addressing effectiveness, acceptability, adherence and behavioral issues. Research, policy, and advocacy agendas, should consider these in the future. 39

140 Poster Discussions Poster Discussion 05: Glycans and Antibody Effector Functions Poster Discussion 04: Behavioral and Social Sciences PD05.0 PD05.02 POSTER DISCUSSIONS Exploring the Influence of N-linked Glycans on the Molecular Dynamics of HIV- gp20 in the Context of Viral Coreceptor Tropism Natasha T. Wood, Elisa Fadda 2, Oliver C. Grant 3, Robert J. Woods 4, Simon A. Travers University of the Western Cape, South African National Bioinformatics Institute, Cape Town, South Africa, 2 National University of Ireland Maynooth, Chemistry, Maynooth, Ireland, 3 National University of Ireland Galway, Chemistry, Galway, Ireland, 4 University of Georgia, Complex Carbohydrate Research Centre, Athens, GA, United States Variable Dependence on Glycan Recognition within a Lineage of VV2-directed HIV Neutralizing Antibodies Nicole A. Doria-Rose, Ryan S. Roark, Penny Moore 2, Michael J. Ernandes, Jinal N. Bhiman 2,3, Chaim A. Schramm 4, Krisha McKee, Sijy O Dell, Mark Louder, Salim S. Abdool Karim 3, Lawrence Shapiro 4, Lynn Morris 2,3, John R. Mascola National Institutes of Health, Vaccine Research Center, NIAID, Bethesda, MD, United States, 2 University of the Witwatersrand and the NICD, Johannesburg, South Africa, 3 Centre for AIDS Programme of Research In South Africa (CAPRISA), Congella, South Africa, 4 Columbia University, New York, NY, United States Background: The dense glycan shield of the HIV- gp20 surface protein protects the virion from recognition and neutralisation by the host immune system, plays a role in binding to target cell chemokine receptors (coreceptors), and forms part of viral epitopes for broadly cross-neutralising antibodies. The composition and distribution of these N-linked glycans also vary depending on the cell-type, stage of infection, and HIV subtype. We have previously applied molecular simulation techniques to show that the spatial dynamics of the gp20 glycoprotein are significantly influenced by the composition of the underlying protein sequence as well as the presence of glycans at specific N-linked glycosylation sites. Methods: To continue this work and further explore the effect of the glycan composition and distribution on HIV coreceptor tropism, we have modelled the structures of three pairs of viral clones from three patients. Each sequence pair consisted of a CCR5- and CXCR4-tropic virus, which had been clonally phenotyped, and represented subtype A, C, and D infections. Oligomannose and complex glycans were linked to N-linked glycosylation sites of each structure based on the reported predominant glycan-type at specific positions along the gp20 glycoprotein. We subsequently applied a molecular dynamics approach, using GROMACS, to identify HIV- coreceptor tropism-associated structural features introduced by glycans on the surface of gp20. Results: Our preliminary results align with our previous work where the presence of glycans had a substantial impact on the dynamics of the V3 loop. Further analysis reveals the degree to which the glycan composition and density around key regions of HIV- gp20 impact the tropism-associated dynamic characteristics of the protein. Conclusions: These results present a unique view on how the glycanprotein, as well as the glycan-glycan, interactions of HIV- gp20 may modulate the infectivity and immunogenicity of the virion. Background: The study of HIV donors with broad and potent neutralizing antibodies can inform rational vaccine design and immunization strategies. We previously identified 2 antibodies from a single lineage in donor CAP256 that target VV2 with breadth up to 60% in clades A and C, and very high potency on many strains. The antibodies varied in dependence on glycan. We sought to isolate additional lineage members and to understand variation in epitope recognition. Methods: Antibodies were isolated via 4-day B cell culture with IL-2, IL- 2, and CD40L. Initial RTPCR was performed using IgG-specific primers. For weeks 9 and 206, wells were re-interrogated using IgA-specific primers. 5 additional antibodies were isolated from week 9 and 4 from week 206, and reconstituted as IgG and IgA. The IgGs were assayed for neutralization on TZM-bl cells on a panel of 49 autologous and heterologous Env-pseudoviruses, as well as viruses with mutations in known epitopes. Results: Several B cell culture wells yielded both IgG and IgA amplicons with the same VDJ sequence, suggesting that class-switching occurred during culture. Nine new antibodies of the CAP256-VRC26 lineage were identified, several of which had breadth similar to the previous best members of the lineage. Two from week 206 lack the characteristic tyrosine sulfation signal, and are also less broadly neutralizing. All depend on residues in V2, with variable and strain-specific effects of glycans. We examined the impact of removing the glycan at N60 which is critical for the PG9 class: some lost neutralization; some were unaffected; and CAP256-VRC26.3, and two nearly identical antibodies cloned independently, showed gain of neutralization, the opposite of PG9. Conclusions: We have expanded the CAP256-VRC26 clonal family. Its members show diversity in recognition of specific determinants within the VV2 epitope, particularly the glycans, which vary in the virus as a means of escape from antibody pressure. 40 HIV Research for Prevention 204 HIV R4P

141 Poster Discussion 05: Glycans and Antibody Effector Functions Thursday, 30 October PD05.03 PD05.04 Polyfunctional Non-neutralizing Fc-antibody Responses in Acute HIV Infection Predict Spontaneous Viral Control Amy W. Chung, Matthew K. Schoen, Hannah Robinson, Anna Licht, Elizabeth Tkachenko, Davey M. Smith 2, Susan J. Little 2, Douglas D. Richman 2, Galit Alter Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 2 University of California San Diego, San Diego, CA, United States Background: Analysis of the moderately protective RV44 vaccine trial revealed that vaccination induced non-neutralizing antibodies (Abs) capable of mediating polyfunctional, highly co-ordinated Fc-effector responses. Additionally, non-neutralizing Fc-effector functions have been associated with delayed disease progression and strength of Fc-effector activity in acute infection correlates with decreased acute viremia. Thus we hypothesized that non-neutralizing Fc-effector functions in acute infection may contribute to spontaneous HIV viral control. Methods: IgG was purified from plasma of acutely infected chronic (CRO) subjects and 9 spontaneous controllers (CTR) at 4, 2, 24 and 48 weeks post infection. Non-neutralizing Fc-activity: Ab dependent cellular cytotoxicity (ADCC), Ab dependent cellular phagocytosis (ADCP), Ab dependent NK cell activation and Ab dependent cellular virus inhibition (ADCVI), along with HIV-specific IgG subclass profiles were characterized across all time points. Results: Despite higher observed ENV-specific bulk IgG titers in CRO subjects, similar levels of ADCC, ADCP and Ab NK activation were detected from the two cohorts. However CTRs developed significantly higher ADCVI activity than CROs at later timepoints. Subclass analysis revealed that ADCVI activity was correlated with ENV-specific IgG3, which was maintained at high levels within CTRs, in comparison to CROs, which progressively lost their IgG3. In addition, CTRs were able to induce highly co-ordinated polyfunctional Fc-effector functions, while in contrast, inversely correlated Fc-responses were detected from CRO subjects at early timepoints. Conclusions: This data suggest that spontaneous controllers of HIV infection are able to induce highly co-ordinated Fc-effector responses, along with maintained high levels of IgG3, that may contribute to HIV control. Thus vaccine strategies able to induce high and sustained levels of polyfunctional Fc-effector responses along with IgG3 may provide protection from and after infection. HIV- Env-specific IgA/IgG Ratio Is Related to Antibody Dependent Cellular Cytotoxicity (ADCC) Responses Observed during Acute/ Early HIV Infection Maria Julia Ruiz, Yanina Ghiglione, Juliana Falivene, María Eugenia Socías 2, Natalia Laufer,3, Pedro Cahn 2,3, Omar Sued 2,3, María Magdalena Gherardi, Horacio Salomon, Ana Maria Rodriguez 4, Gabriela Turk Instituto de Investigaciones Biomédicas en Retrovirus y Sida, Universidad de Buenos Aires/CONICET, Buenos Aires, Argentina, 2 Fundación Huesped, Buenos Aires, Argentina, 3 Hospital Juan A. Fernández, Unidad Enfermedades Infecciosas, Buenos Aires, Argentina, 4 Instituto de Inmunología, Genética y Metabolismo (INIGEM), Universidad de Buenos Aires-CONICET, Buenos Aires, Argentina Background: Immune correlates analysis of the RV44 vaccine trial revealed that high levels of Env specific IgA antibodies (ab) may have mitigated the capacity of protective vaccine-induced IgG ab to mediate ADCC. Aim: To evaluate the relation between Env-specific IgG and IgA ab and ADCC responses in the context of acute/early primary HIV infection (PHI) Methods: Plasma from 20 HIV + subjects enrolled during PHI, obtained at enrollment (baseline sample) and year post-infection sample (ypi), 0 HAART-naïve chronically infected patients (C) and 7 Elite Controllers (EC) were used. Percentage of ADCC-killing was determined using the rapid and fluorometric ADCC assay. Env-specific IgG and IgA ab were assessed by ELISA. Data was analyzed using non-parametric statistics Results: Levels of Env-specific IgG ab were significantly higher in PHI ypi sample compared to baseline (median 2802 vs ng/ml p=0.006) and as expected directly correlated with %ADCC-killing both at baseline (r=0,476 p= 0,039) and ypi samples (r=0,627 p=0,06). Similar direct correlations were observed in C and EC (r=0,898 p=0,033 and r=0,766 p=0,02 respectively). Conversely, IgA titers declined over the course of acute infection (baseline vs ypi: 200 vs 35 respectively, p=0,00) and did not correlate with ADCC responses observed at the ypi sample. Envspecific IgG/IgA ratios were also significantly higher at the ypi sample compared to baseline (median 556,5 vs 0,39, respectevly, p= 0,000). More interestingly, Env-specific IgG/IgA ratios directly correlated with %ADCC-killing at the ypi sample (r=0,538 p=0,047) Conclusions: The direct correlation between IgG/IgA ratio and %ADCCkilling observed at the ypi sample indicate that Env-specific IgA ab might be interfering with the magnitude of IgG-mediated ADCC responses in subjects undergoing PHI. This result indicates that the elicitation of Env-specific IgA ab hampers protective ADCC responses not only in vaccinees but also in natural infection and should be taken into account in vaccine settings POSTER DISCUSSIONS 4

142 Poster Discussions Poster Discussion 05: Glycans and Antibody Effector Functions PD05.05 POSTER DISCUSSIONS Fcγ Receptor Functional Variability Impacts on Mother-to-Child Transmission of HIV- Ria Lassauniere,2, Glenda E. Gray 3, Louise Kuhn 4, Caroline T. Tiemessen,2 National Institute for Communicable Diseases, Centre for HIV and STIs, Johannesburg, South Africa, 2 University of the Witwatersrand, Faculty of Health Sciences, Johannesburg, South Africa, 3 Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital, Soweto, South Africa, 4 Gertrude H. Sergievsky Centre, College of Physicians and Surgeons and Department of Epidemiology, Mailman School of Public Health, Columbia University, New York, NY, United States Background: The study model of maternal-infant HIV- transmission provides insights into what may constitute protective immunity to HIV-. Using this model, we investigated the potential role of gene variants known to influence Fc gamma receptor (FcγR) mediated effector mechanisms that include antibody-dependent cell mediated cytotoxicity and phagocytosis. Methods: We genotyped FcγR functional variants and gene copy number in 207 HIV- infected mothers and their infants: 38 were non-transmitting mothers and their exposed-uninfected infants; 69 transmitting mothers and their intrapartum or in utero infected infants. The following functional variants were investigated: FcγRIIa-p.H3R, FcγRIIb-p.I232T, FcγRIIc-p.T8I, FcγRIIIa-p.F58V, FcγRIIIb-HNAa/ HNAb/HNAc, FCGR2B/C promoter variants, and genetic markers predicting FcγRIIc expression. Results: Mothers carrying at least one FcγRIIIa-58V allele had a 68% reduction in the odds of transmitting HIV- in utero (OR 0.38, 95%CI: , P = 0.002) compared to mothers who did not carry a 58V allele. When mothers and infants possessed discordant genotypes, in utero transmission of HIV- was associated with the FcγRIIIa-58FF genotype in the mother (OR: 2.959, 95%CI: , P = 0.02), while in utero acquisition was associated with the FcγRIIIa-58FV genotype in the infant (OR 2.27, 95%CI: , P = 0.039). In concordant mother-infant pairs, an increased odds of HIV- transmission/acquisition in utero was independently associated with homozygosity for the FcγRIIb-232T allele (OR 3.208, 95%CI:.0-0.6, P = 0.048) and the presence of at least one FcγRIIIb-HNAc allotype (OR 2.37, 95%CI:.-5.08, P = 0.035). Conclusions: Antibody affinity variability conferred by the FcγRIIIa- F58V allele, and lower inhibitory activity of FcγRIIb in both mother and infant was associated transmission and acquisition of HIV-. These findings suggest a role for FcγR-mediated effector functions in transmission and acquisition of HIV-. 42 HIV Research for Prevention 204 HIV R4P

143 Poster Discussion 06: Policy, Advocacy and Modeling Thursday, 30 October PD06.0 PD06.02 To Use or Not to Use Clade Specific Vaccines: A Central Question Dobromir Dimitrov, James Kublin, Scott Ramsey, Larry Corey Fred Hutchinson Cancer Research Center, Seattle, WA, United States Background: Recent studies support the hypothesis that immune responses to HIV- are markedly influenced by clade. Vaccines currently entering efficacy trials are clade C. We discuss whether these vaccines should be used in other regions before clade-specific prototypes are constructed and evaluated. Methods: Mathematical models were used to simulate the impact of vaccination strategies on the HIV epidemics in South Africa (SA) and San Francisco (SF). We compared three interventions based upon a 50% vaccine efficacy (VE) of a clade-specific vaccine, reduced to 20-40% VE in non-clade matching regions: i) immediate use of non-matching vaccine; ii) delayed intervention by developing a clade-specific vaccine and iii) immediate use of non-matching vaccine replaced by a clade-specific vaccine when developed. Results: While implementation of a maximally effective vaccine provides the greatest population benefits, immediate vaccination with a 20-40% VE may prevent thousands of new infections in SF and millions in SA over 30 years reducing the HIV prevalence by up to 7 %. Vaccination with 50% VE delayed for 5 years needs 6 and 2 years to break-even with immediate vaccination with 20 and 30% VE, respectively, but is not able to match the 30-year impact of immediate vaccination with 40% VE. Replacing a vaccine with 30% VE with 50% VE vaccine after 5 years prevents 5% more infections than delayed vaccination with 50% VE but these benefits may be lost if vaccinees reduce condom use. Conclusions: The immediate use of a non clade matching HIV vaccine may be desirable if it does not lead to changes in sexual behavior that result in increased HIV infection risk. The pursuit of higher VE through clade matching prototypes will hasten the reduction in HIV prevalence over time if the initial vaccination program does not impact adherence to subsequent vaccination programs. This study illustrates the importance of developing biomarkers of HIV- acquisition to support more rapid and economical assessment of clade specific vaccines. The Tipping Point: Moving from Rhetoric to Real Milestones for Ending AIDS Emily Bass, Emily D. Donaldson, Mitchell Warren, Kevin Fisher AVAC, New York, NY, United States Background: The epidemiologic tipping point in the AIDS epidemic is the point at which the rate of ART treatment scale-up outpaces HIV incidence. AVAC and amfar analyzed modeling research and consultated with top HIV prevention experts to lay out essential steps that must be taken by national governments, international organizations, civil society, researchers and technical agencies to steadily reduce annual new HIV infections and continue to expand HIV treatment access in order to meet the tipping point. Methods: The analysis used modelling to build a prevention advocacy agenda around ending AIDS. The targets set reflect best-case scenario calculations based on published modeling and epidemiologic data, as well as analysis provided by experts in the field. Data projections were cross-checked with modelers and epidemiologists. Modelling data is tracked and updated to ensure the most recent metrics are used, and real-time data is included and analyzed as available. Results: By engaging with modelling data, advocates are able to set real, measurable targets and hold accountable national and global agencies to ensure key data points are met, including decreasing new HIV infections and deaths, as well as specific epidemiologic and policybased milestones tied to global scale-up of critical interventions. Rate of treatment coverage and the rate at which incidence decreases are key to the analysis thus, real-time assessment of results allows advocates to use informed decision-making to ensure these time-related targets kept on track. Conclusions: By using data-driven targets, such as the tipping point, as an advocacy tool, advocates can translate complex models for policy makers and donors to ensure continued and timely scale-up of core interventions and sustained resources. It is essential to continue scale-up and funding of key interventions and research towards ending AIDS, establishing a set of targets that advocates can track globally and nationally ensures movement towards, and beyond, the tipping point. POSTER DISCUSSIONS 43

144 Poster Discussions Poster Discussion 06: Policy, Advocacy and Modeling PD06.03 PD06.04 LB POSTER DISCUSSIONS Bioethical and Biostatistical Considerations of Innovative HIV Prevention Trial Designs Darpun D. Sachdev,2, Ryan Whitacre 3, Susan P. Buchbinder 2,4 Center for AIDS Prevention Studies (CAPS), UCSF, San Francisco, CA, United States, 2 San Francisco Department of Public Health, Bridge HIV, San Francisco, CA, United States, 3 University of California at San Francisco, San Francisco, CA, United States, 4 University of California at San Francisco, Department of Epidemiology and Biostatistics, San Francisco, CA, United States Background: Given the efficacy of pre-exposure prophylaxis (PrEP), there is ongoing debate regarding whether future placebo-controlled biomedical HIV prevention trials are ethically justifiable. Several innovative trial designs under consideration to test new biomedical HIV prevention modalities provide PrEP as part of the standard of prevention or use PrEP as an active control, however, little is known regarding the ethical considerations of these designs. Methods: We purposively sampled and conducted in-depth interviews with 2 biostatisticians and 3 bioethicists, each with >0 years of experience in HIV prevention trials. We sought to understand potential HIV vaccine or long-acting PrEP trial designs that were under consideration and preferences toward the designs. We analyzed interview transcripts using constant comparative methods to inductively develop and refine themes. Results: Several innovative HIV prevention trial designs were elicited including factorial designs, pre-randomization PrEP run-in periods, noninferiority studies and adaptive trials. Respondents agreed that PrEP should be offered in future trials unless there is ethical justification precluding inclusion. Adaptive trial designs, which allow for modification of a trial in response to study data, were highly acceptable to both biostatisticians and bioethicists due to their potential to: ) reassess clinical equipoise during the course of the study, 2) utilize informed consent to apprise participants of prespecified trial design changes, and 3) save time and resources while also assessing the number of clinical endpoints necessary for regulatory approval. Conclusions: Adaptive trial designs may allow for scientifically rigorous studies to evaluate the efficacy of new biomedical HIV prevention modalities while also maximizing benefits to trial participants. Ongoing interviews with investigators and regulators will evaluate for the real world limitations of these designs. Differences in PrEP Knowledge and Use in U.S. MSM Users of a Popular Sexual Networking Site Surveyed in August 203 and January 204 Kenneth H. Mayer,2, Catie Oldenburg 3,4, David S. Novak 5, Douglas Krakower,6, Matthew J. Mimiaga,4,7 Fenway Health & Harvard University, The Fenway Institute, Boston, MA, United States, 2 Beth Israel Deaconess Medical Center, Infectious Disease Division, Boston, MA, United States, 3 Fenway Health, The Fenway Institute, Boston, MA, United States, 4 Harvard School of Public Health, Epidemiology, Boston, MA, United States, 5 On-Line Buddies, On-Line Buddies Research Institute, Cambridge, MA, United States, 6 Beth Israel Deaconess Medical Center, Division of Infectious Diseases, Boston, MA, United States, 7 Massachusetts General Hospital / Harvard Medical School, Psychiatry, Boston, MA, United States Background: Pre-exposure prophylaxis (PrEP) has been recommended for HIV prevention for at risk men who have sex with men (MSM). Prior reports have suggested slow uptake. Methods: U.S. MSM members of a sexual networking site were invited to complete a survey in August, 203 (Wave : W) and January, 204 (W2) that included a variety of questions related to HIV prevention practices. Comparisons of those who responded at each time point were conducted using Fisher s exact tests in order to assess differences in PrEP knowledge and uptake between W and W2. Study design did not permit assessment of whether MSM responded once or at both time points. Results: Although more MSM responded in W than W2 (6,683 vs. 4,759), they were similar in median age (45 y.o.), race/ethnicity (about 86% White), educational attainment (about 2/3 had at least a college degree) and regional geographic distribution (almost ¼ from the Southeast, with the next largest numbers being in the Northeast, Great Lakes region or Midwest-about 5% each). The majority reported engaging in condomless anal sex (CAS) in the prior 3 months (58.7% in Wave and 74.% in Wave 2). A higher proportion of MSM in W2 had heard of PrEP than in W (43.8% vs. 27.3%, p< 0.00). Among MSM reporting CAS in the prior 3 months, PrEP interest increased between W and W2 (54.% vs. 60.4%, p=0.002) and use also increased (2.0% vs. 3.%, p=0.004). PEP knowledge increased (38.9% and 49.7% in W and W2, p< 0.00), but PEP utilization among MSM reporting CAS in the prior 3 months did not significantly increase (4.3% in W and 4.7% in W2, p=0.44). Conclusions: Although recent increases in PrEP interest and use among U.S. MSM accessing a sexual networking site were detected, the majority of those who could benefit have not availed themselves of this prevention modality, with the majority of at risk respondents not being knowledgeable about PrEP. Further educational efforts are needed for U.S. MSM about PrEP. 44 HIV Research for Prevention 204 HIV R4P

145 Poster Discussion 06: Policy, Advocacy and Modeling PD06.05 LB Thursday, 30 October Guidelines on Post Exposure Prophylaxis for HIV: Recommendations for a Public Health Approach Nathan Ford HIV/AIDS, World Health Organization, Geneva, Switzerland WHO has recently updated recommendations on the use of antiretroviral drugs as post exposure prophylaxis (PEP) to prevent HIV infection. The guidelines are based on a public health approach that considers feasibility and effectiveness across a variety of settings. In producing these guidelines the key principles of availability, acceptability, accessibility and quality have been considered. In contrast to previous published WHO guidelines, these guidelines consider all exposures and provide recommendations for all populations. In doing so no distinction between occupational and non-occupational settings has been made with the aim of simplifying guidance for PEP prescribing. These guidelines promote the approach that individuals should be offered PEP if the exposure constitutes a significant risk of transmission and the same drug regimen should be prescribed irrespective of exposure source. Recommendations for preferred regimens, simplifying prescribing approaches and supporting adherence are also provided. POSTER DISCUSSIONS 45

146 Overview of Poster Sessions Posters Presented on Wednesday, 29 October P0: Acute Seroconversion P02: Community Engagement and Advocacy P03: Correlates of Protection and Exposure P04 : Correlates of Protection in Highly Exposed Seronegative People P05: Family Planning P06: Financial Incentives P07: Good Participatory Practices and Community Involvement P08: HIV Care P09: HIV Testing and Counseling P0: Immunogens P: Informed Consent P2: Innate Immunity P3: Key Populations P4: MPT Development P5: Novel Formulations, Agents and Microbicides P6: Passive Antibody Functions P7: Post-exposure Prophylaxis P8: Preclinical Evaluation of Vaginal Films and Gels P9: PrEP Implementation P20: PrEP: Resistance, Modeling and Acceptability P2: Product Acceptability and Adherence P22: Resistance and Treatment Failure P23: Retention and Adherence in Trials P24: T-Cell (CD4 & CD8) Responses P25: Transmission and Viral Diversity P26: Vaccine Clinical Trials HIV Research for Prevention 204 HIV R4P

147 Overview of Poster Sessions Posters Presented on Thursday, 30 October P27: Adjuvants P28: Behavioral and Social Sciences P29: Circumcision and Acceptability P30: Condoms: Attitudes, Use and How to Increase P3: Drug Transporters P32: Ethics and the Law P33: Evaluation of Novel Compounds in Cell-Based Systems P34: Glycans and Antibody Effector Functions P35: HIV Drug Resistance in vitro P36: HIV Incidence and Prevalence P37: Immunogenetics P38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring P39: Molecular Epidemiology P40: Mucosal Immune Activation and Inflammation P4: Novel Vaccine and Prevention Concepts P42: Participation in Trials: Willingness, Benefits and Challenges P43: Policy, Advocacy and Modeling P44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P45: Pregnancy and Prevention of Mother to Child Transmission P46: PrEP Trials: Preparing for Demos, Participant Experiences P47: Resource Tracking and Economics P48: Risk Perception P49: Sexually Transmitted Infections P50: Tenofovir Gel: Acceptability and Adherence P5: Therapeutic Vaccine and Viral Latency P52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV P53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora

148 Posters Posters 0: Acute Seroconversion P0.0 P0.02 Recall of Acute Retroviral Symptoms in a Multicentre Cohort Study in Africa Eduard J. Sanders,2, Kimberley A. Powers 3, Etienne Karita 4, Anatoli Kamali 5, William Kilemba 6, Susan Allen 7, Eric Hunter 7, Omu Anzala 8, Pat Fast 9, Matthew A. Price 9,0 KEMRI-Wellcome Trust Research Programme, Kilifi, Kenya, 2 University of Oxford, Headington, United Kingdom, 3 University of North Carolina, Chapel Hill, NC, United States, 4 Project San Francisco, Kigali, Rwanda, 5 Medical Research Council/Uganda Virus Research Institute, Entebbe, Uganda, 6 Zambia Emory Research Project, Lusaka, Zambia, 7 Emory University, Atlanta, GA, United States, 8 Kenya AIDS Vaccine Initiative Institute of Clinical Research, Nairobi, Kenya, 9 International AIDS Vaccine Initiative, New York, NY, United States, 0 University of California at San Francisco, San Francisco, CA, United States Background: Remarkable differences exist for reported acute retroviral symptoms (ARS) in African adults when acquiring HIV- infection, notably for fever (range: 9-78%), or diarrhea (range: 2-42%). This may lead to under-appreciation of the clinical presentation and delayed diagnosis of patients with acute HIV- infection (AHI). We sought to determine whether reporting of ARS was associated with timing of ascertainment of signs and symptoms and clinical research centre (CRC) at nine collaborating CRC s in Africa. Methods: Adults with acute or early HIV- infection in a multicenter HIV- incidence study were enrolled in a sub-study assessing ARS. Estimated date of infection (EDI) was based on a positive plasma viral load or p24 antigen test prior to seroconversion, or the mid-point between a negative and positive HIV- serologic test. ARS signs and symptoms were assessed at sub-study enrollment. We used log-binomial regression to estimate the prevalence of ARS signs and symptoms by recall period (i.e. period = enrollment 42 days after EDI and period 2 = enrollment > 42 and 90 days after EDI) and CRC. Results: Among a total of 24 and 8 participants evaluated in period and 2, respectively, 66.0 (95% confidence interval (CI): %), and 49.4 (95% CI: %, p=0.0006) reported having had at least one sign or symptom. The most common symptoms of ARS were fever, headache, myalgia/arthralgia, and fatigue. With the exception of fever, diarrhea, and oral ulcers, the prevalence of all signs or symptoms was statistically significantly greater in those who were evaluated in period as compared to those evaluated in period 2 (Figure). There was considerable variability in the prevalence of signs and symptoms by CRC. Conclusions: In this multicenter African cohort, the prevalence of ARS symptoms of patients evaluated within 6 weeks following EDI was higher for all symptoms except for fever, diarrhea and oral ulcers as compared to patients who were evaluated between 6 weeks and 3 months of the EDI. Long-term Follow up of HIV Seroconverters from the VOICE Trial - An Analysis of Data from MTN-05 Sharon Riddler, Marla Husnik 2, Arendevi Pather 3, Thesla Palanee 4, Gonasagrie Nair 5, Felix Mhlanga 6, Marthinette Taljaard 7, Clemensia Nakabiito 8, Anamika Premrajh 3, Lisa Levy 9, Vanessa Elharrar 0, Gita Ramjee 3, Jennifer Balkus,2, MTN-05 Protocol Team University of Pittsburgh, Pittsburgh, PA, United States, 2 Statistical Center for HIV/AIDS Research & Prevention (SCHARP), Seattle, WA, United States, 3 Medical Research Council, Durban, South Africa, 4 Wits Reproductive Health and HIV Institute, University of the Witwatersrand, Johannesburg, South Africa, 5 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa, 6 University of Zimbabwe - University of California San Francisco Collaborative Research Programme, Harare, Zimbabwe, 7 The Aurum Institute, Klerksdorp, South Africa, 8 Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 9 FHI 360, Research Triangle Park, NC, United States, 0 Division of AIDS, NIH, Bethesda, MD, United States, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 University of Washington, Seattle, WA, United States Background: In 2008, the Microbicide Trials Network (MTN) initiated a multi-site, long-term observational study of women who become HIV infected during microbicide trials participation to assess the impact of investigational products on HIV disease progression, treatment response, and clinical and behavioral outcomes (MTN-05). Here we report on women enrolled from the VOICE trial. Methods: MTN-05 recruited eligible VOICE participants from 5 sites in South Africa, Zimbabwe and Uganda. Visits occurred at enrollment and at, 3, 6 months after detection of HIV infection, then every 6 months thereafter. CD4+ T cell count, HIV- RNA, clinical history, and risk reduction counseling are completed at each visit. We evaluated the effect of the interventions (oral TDF, oral FTC/TDF, and vaginal tenofovir gel) on HIV disease progression using Cox proportional hazard models to assess time to CD4< 350, ART initiation, and first AIDS-defining event as separate outcomes. Results: MTN-05 enrolled 255/356 (72%) of VOICE seroconverters; 93% were from South Africa. Median age at MTN-05 enrollment was 23 years; median time from detection of HIV infection to enrollment was 2. months. To date, median follow-up is 22.9 months (cumulative 457 person-years). At enrollment, median CD4+ and HIV- RNA were 539 cells/mm 3 and 4.4 log 0 copies/ml. We observed no significant difference by VOICE study arm with regard to time to CD4< 350, ART initiation, or AIDS-defining events. ART was initiated by 49 women (3 prior to MTN-05 enrollment); 82% met local CD4 guidelines for ART initiation and 6% started ART due to pregnancy. Initial ART regimens were NNRTI based, most commonly EFV/TDF/3TC (6%); only 2 (4%) included d4t. AIDS-defining events occurred in (4%) women. Conclusions: Long term follow-up of participants who seroconvert in prevention trials is feasible, acceptable and is an important component of the assessment of ART-based prevention modalities. 48 HIV Research for Prevention 204 HIV R4P

149 Posters 0: Acute Seroconversion Wednesday, 29 October P0.03 LB VOICES: A Qualitative Study with MSMs in South Florida May 204 Gregory Edwards, William Green 2, Marsha A. Martin 3,4, William DiStefano 5, Leonard Jones 6 Flowers Heritage Foundation, Oakland, CA, United States, 2 Broward, Health Care Services Section, Broward County, FL, United States, 3 Get Screened Oakland, Office of the Mayor, Oakland, CA, United States, 4 Urban Coalition for HIV/AIDS Prevention Services, Washington, DC, United States, 5 Get Screened Oakland, Oakland, CA, United States, 6 Human Services Section-Ryan White Part A, Broward County, FL, United States Background: In Broward County, FL 2,00 persons lived with HIV disease in 20, and 3,334 (5.8%) were unaware, with the largest number in Ft. Lauderdale. More than 000 new cases of HIV were reported, an increase of 25% from 200. New cases affected persons between the ages of 40 and 49. MSMs make up approximately 70% of all infections. Methods: A multifaceted approach assessed the epidemic among gay, bisexual, and other MSMs: community advisory workgroup, literature review, 25 key informant interviews, 7 focus groups with men of known and unknown HIV serostatus, 4 focus groups with transgender women and consumer surveys. The study sought to answer the following question: What are the socio-sexual-cultural circumstances that lead to increased risk for and transmission of HIV among gay, bisexual men and transgender women? Results: Young and older MSMs report and experience social isolation, disaffiliation and limited choices for socialization and healthy relationships. HIV infection is felt to be just a matter of time. For some gay men who have been living with HIV for more than 5 years, HIV messages are not relevant to their experiences. The availability and accessibility of venues and multi-day circuit parties that promote sexual play and risk-taking behaviors also contribute to the spread of HIV among gay and bisexual men, and transgender women. Online hookups that advertise bareback sex play create the perception that it is no longer a socio-sexual cultural norm to use protective barriers during sexual intercourse. Gay, bisexual MSMs living with HIV, many of whom experience shame, low self-esteem and isolation have very limited venues for positive self expression and health promotion. Conclusions: The Broward County HIV epidemic is influenced by sociosexual-cultural economic dynamics: anonymous sex, alcohol, and crystal meth used to ease hardships associated with a lack of social and communications skills, loneliness, isolation, and exhaustive vigilance create the perfect storm of HIV infection. 49

150 Posters Posters 02: Community Engagement and Advocacy P02.0 P02.02 Managing Communication and Facilitating the Exchange of Ideas in a Large, Multi-site Clinical Trial: Experiences from the ASPIRE Study Rachel Scheckter, Katie Schwartz, Ashley Mayo, Jared Baeten 2, Thesla Palanee 3, Lydia Soto-Torres 4, Kristine Torjesen FHI 360, Durham, NC, United States, 2 University of Washington, Seattle, WA, United States, 3 Wits Reproductive Health and HIV Institute (WRHI), Johannesburg, South Africa, 4 National Institute of Allergy and Infectious Diseases, Bethesda, MD, United States Background: Streamlined communication and coordinated strategies are essential for large multi-site clinical trials to ensure high-quality implementation and to minimize burden from large volumes of disaggregated input across multiple protocol team partners. Methods: A review of approaches to improve coordination and enhance communication in the MTN-020 study (ASPIRE) was conducted to develop qualitative and quantitative descriptions of these activities. Results: Multiple strategies have been used in ASPIRE to streamline communication and coordination across sites and the protocol team. Six in-person trainings, designed to incorporate adult learning techniques and establish common approaches, were conducted prior to study start. Additional workshops and trainings have continued throughout implementation and 38 assessment visits have been conducted to date. Assessment visits focus on strengthening site-specific systems and communication strategies. A written study communication plan was developed at study outset that included use of weekly priority s with sites to minimize communication burden of important information and action items. Communication was streamlined through monthly or twice-monthly implementation calls with individual site leaders and weekly management team calls to share key updates. Cross-site learning was fostered through cadre-specific listservs, semiannual in-person meetings, monthly site-led protocol team calls, and quarterly calls to strengthen adherence counseling and qualitative data collection. The ASPIRE communication strategy has led to sharing of innovative sitedriven approaches, such as participant engagement strategies, as well as early identification and quick resolution of implementation issues, such as participant and community myths and misconceptions. Conclusions: Strategies to streamline communication and coordination in ASPIRE to improve efficiency and quality of study implementation could inform implementation of other large multi-site clinical trials. A Rectal Revolution Takes a Village: Developing an Educational Video about Rectal Microbicides Clare Collins, Barbara Friedland 2, Jim Pickett 3 Microbicide Trials Network, Pittsburgh, PA, United States, 2 Population Council, New York, NY, United States, 3 International Rectal Microbicide Advocates, Chicago, IL, United States Background: Rectal microbicides (RMs) are new products being developed to prevent HIV transmission through receptive anal intercourse. Given that most people are unfamiliar with RMs, educational materials that are accessible, understandable and appealing to multiple communities are essential. To address this need, the Population Council, International Rectal Microbicide Advocates (IRMA) and the Microbicide Trials Network (MTN) developed a 4-minute video on RMs, The Rectal Revolution is Here: an Introduction to Rectal Microbicide Clinical Trials, in conjunction with the launch of an international Phase II RM clinical trial. Methods: The video, which combines animation and documentarystyle footage, was developed with the guidance of a Video Advisory Committee (VAC), comprised of 2 individuals from international trial sites, non-affiliated researchers, and IRMA advocates. The VAC provided input and expertise on all aspects of the video, from script content to translations to final edit. An early cut of the video was viewed by over 80 participants at Microbicides 202 who gave feedback via a survey, and was tested in 3 focus group discussions at five sites in English, Spanish, and Thai among more than 00 gay men, other men who have sex with men, and transgender women. Results: Feedback from the surveys and focus groups resulted in several significant changes to clarify key messages, including the addition of information about HIV testing, the cutting of a main character as well as some interview footage, and a change in music. The final video has been made available to trial sites and advocates, and online via YouTube. Conclusions: The extensive year-long process with multiple inputs used to develop the video resulted in concrete suggestions to improve it, and to make it relevant and accessible for intended audiences. By incorporating this vital feedback, video producers were able to develop a culturally competent educational video that was inclusive and responsive, and is being used extensively. 50 HIV Research for Prevention 204 HIV R4P

151 Posters 02: Community Engagement and Advocacy Wednesday, 29 October P02.03 Seven Steps to Strengthen Community Engagement in HIV Preventions Trials in Durban P02.04 How Community Education Tools Facilitated Understanding of the ASPIRE Vaginal Ring Study: Kampala Experience Neetha Shagan Morar South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: Effective community engagement is the cornerstone of cultivating ownership, trust and acceptance of research within communities. Researchers at the six sites of the HIV Prevention Research Unit (HPRU) in Durban interact with a diverse range of urban and semirural communities where trials are conducted. We describe the outcomes of the seven steps in community engagement while conducting microbicide and other HIV prevention trials at HPRU. Methods: Using a 7 step cyclical process, community engagement began a decade ago at the research sites. The process involved: () conducting a situational analysis at entry; (2) education and outreach; (3) networking and partnerships with stakeholders; (4) selection of community advisory boards (CABs); (5) input on protocol and study procedures; (6) participant engagement and (7) dissemination of study outcomes. Results: Community partnerships with stakeholders and CABs have been sustained for over a decade. Ongoing engagement fostered trusting relationships and facilitated acceptance of trials by community and participants. Education from stakeholders on community culture and social dynamics helped researchers address community perceptions of HIV prevention and microbicides. CAB members observed the destruction of blood samples and this transparency enhanced their understanding of ethics in research and dispelled the myth that researchers sold blood. Trial participants volunteered to publicly share their experiences during the dissemination of trial results to stakeholders. They joined CABs and are currently advocates of HIV prevention research by promoting adherence and retention. Conclusions: Stakeholders, CABs and trial participants assisted in developing trust between communities and researchers, who were transparent and engaged them throughout the research lifecycle. Education and information enabled stakeholders to make informed decisions and facilitated their understanding of trial results and acceptance of the conduct of trials in their communities. Patrick Ndawula, Teopista Nakyanzi, Juliane Etima, Samuel Kabwigu, Flavia K. Matovu, Sophie C. Nanziri, Doreen Kemigisha, Stella Nanyonga, Rhonda White 2, Cheryl Cokley 2, Clemesia Nakabiito Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 FHI 360, Duham, NC, United States Background: UNAIDS Good participatory practice guidelines for biomedical HIV prevention trials recommends that sufficient trial information, such as study objectives, procedures, risks, benefits, and what is expected of participants, is provided for potential participants to make informed decisions. We describe how ASPIRE community education tools increased study awareness and enabled literacy regarding research, reproductive health, family planning, and HIV prevention. Methods: Kampala site began community education about ASPIRE in July 202. Community education tools are used to raise awareness and knowledge about the study. Tools like the ASPIRE Community Education flip chart and fact sheets; contain graphic images to support learning in low literacy populations. Visual aids, such as IUCD and pelvic models and a demonstration vaginal ring, are also used. These tools have helped to dispel anxieties and rumors, provide accurate information, and reduce potential barriers to participation. Following sensitization sessions in which these educational tools are used, a comprehension assessment is administered to randomly selected session attendees. The assessment is invaluable in identifying information that study educators need to clarify to enable informed decision-making. For example, when assessment results identified limited understanding of the placebo concept, educators adjusted their messaging to improve learning. Results: Through March 204, 7,6 women were educated about ASPIRE; 6,03 (79%) completed a comprehension assessment. 5,893 (98%) responded that one should be HIV negative to participate in the study; 5,592 (93%) identified 3 HIV prevention methods; (8%) indicated the study product to be tested; and 4,690 (73%) named 2 reliable family planning methods. However only,924 (32%) explained the placebo concept. Conclusions: Community educational tools are effective in facilitating potential participants understanding of research, reproductive health, and family planning and HIV prevention. 5

152 Posters Posters 02: Community Engagement and Advocacy P02.05 Health Science Journalists Are Bridge Partners as HIV Prevention Research Moves Ahead Bobby Ramakant, Badri N. Saxena 2,3 Citizen News Service (CNS), Lucknow, India, 2 Microbicides Society of India, Delhi, India, 3 ICMR Microbicides Expert Group, Government of India, Department of Health Research, Delhi, India P02.06 Bridging the Gap: Uniting Participants and Staff for Improved ASPIRE Study Metrics at Wits Reproductive Health and HIV Institute in Hillbrow Krishnaveni Reddy, Helen Rees, Thesla Palanee Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: Health science journalists routinely focus on event reporting news and seldom get opportunity to dwell deep into health science research, understand science as it moves ahead. Media owners also are reluctant to invest in opportunities that further understanding of their health science journalists and editors. Methods: CNS supported by its partner Microbicides Society of India (MSI) worked with CNS Correspondents team and identified those interested in HIV prevention research and/or were based in countries where HIV prevention research is moving forward. CNS Annual Health Fellowship programme organized audio/ video calls with CNS Fellows every month on specific health science research issues and Fellows interviewed incountry experts and regional/ global experts to develop in-depth news feature articles to keep readers abreast of progress on HIV prevention research. CNS Correspondents and Fellows also joined AVAC or IRMA webinars whenever appropriate to further their own understanding on key developments in HIV prevention science. CNS also supported its correspondents to go to HIV or other related conferences in countries, regions or globally to do key informant interviews with scientists who were doing HIV prevention research, other advocates, trial participants, among others and develop these news feature articles published and syndicated through CNS. Results: Over 00 HIV prevention research scientists, advocates, trial participants were interviewed by CNS Correspondents during and articles got published and syndicated by CNS to online and print media in Asian and African nations. These news feature articles were also disseminated through social media platforms, conference websites, official conference newspapers, among others. Conclusions: - important for media house owners to invest in opportunities for health science journalists to understand scientific research and follow it consistently - important for media to be involved as key partners of HIV prevention research Background: ASPIRE is a Phase 3 study assessing safety and effectiveness of an intravaginal Dapivirine ring in preventing HIV infection. At Wits RHI, we recognize that to improve product adherence and visit retention metrics, understanding the importance of meeting study objectives and being committed to the HIV prevention agenda must be optimized amongst both participants and staff. To achieve this, novel strategies were implemented to secure participant and staff buyin. Methods: Standard practice for encouraging adherence and retention entails staff counseling participants regarding these metrics at study visits followed by telephonic retention/adherence reminders. Although effective, we note that achieving study metrics is not a concern for most participants and routine visit contact is insufficient to develop thorough study objective understanding and build commitment and trust needed to impact these metrics positively. We have found that study understanding, commitment and shared study metric accountability can be encouraged among participants and staff by frequent professional but social interaction. Informal participant-staff engagement activities (e.g. tea/retention parties, movie days) are hosted on visit-free days with the aim of bridging the communication gap and establishing a shared commitment to the ASPIRE team- goal motto. Discussions include prior and current trial information, study objectives, adherence and retention progress and participants contributions to these metrics. Results: These activities remind participants of their impact on the HIV prevention field and their health. The non-judgmental atmosphere results in engaged discussion, disclosure of concerns and addressing critical issues. Participants develop support networks and peer-educate each other during visit waiting times. Conclusions: These activities have facilitated building of participant-staff rapport and potentially increased study commitment. Consequently, communication, product adherence and retention are enhanced. 52 HIV Research for Prevention 204 HIV R4P

153 Posters 02: Community Engagement and Advocacy Wednesday, 29 October P02.07 Engaging the Community and Stakeholders to Develop Microbicide Communication Materials in Kenya Elizabeth Ryan, Emily Bockh, George Githuka 2, Caroline Mackenzie 3, Alice Olawo 3, A. Cornelius Baker, Elizabeth Tolley 4 FHI 360, Washington, DC, United States, 2 Ministry of Health, Nairobi, Kenya, 3 FHI 360, Nairobi, Kenya, 4 FHI 360, Durham, NC, United States P02.08 Factors that Influence Trainees Acquisition of Knowledge about Biomedical HIV Prevention Research: Experience from the Field Oluwatosin Bamidele Alaka, Morenike Oluwatoyin Ukpong 2, Florita Durueke, Augustina Obiajulu Amuamuziam New HIV Vaccine and Microbicide Advocacy Society, Programmes, Lagos, Nigeria, 2 New HIV Vaccine and Microbicide Advocacy Society, Coordinator / Programmes, Lagos, Nigeria Background: In 200 CAPRISA 004 provided proof of concept that peri-coital, vaginal use of tenofovir % microbicide gel can reduce HIV acquisition among women. Communication campaigns will play a key role in generating demand and educating women on correct use. Communicating about Microbicides with Women in Mind used a participatory process to create and pretest a suite of communication materials for potential users, partners and health care providers in Kenya. The project is part of a larger USAID-funded effort to support microbicide introduction if/when tenofovir gel is proven effective. Methods: The project conducted numerous engagement activities. A national policy consultation was held to identify priority audiences for microbicide communication in Kenya. A Project Advisory Committee (PAC), led by the National AIDS Control Programme and consisting of local stakeholders, was formed. Audience consultations were conducted with young women, female sex workers (FSWs), women in relationships, and men, and were followed by a national message development workshop (MDW) to draft messages and materials. Participants from the policy consultation and MDW were reconvened to review research results, discuss final material changes, and provide input into a communication strategy and materials adaptation guide. Results: Engagement activities resulted in a well-received and effective suite of materials, including logos, posters, radio spots, animated TV storyboards, a brochure, counseling algorithms, and educational flipcharts. These materials were refined through two round of pretesting and a study to assess their impact on microbicide interest, attitudes and condom migration. The PAC provided substantive contributions throughout. Conclusions: Stakeholder engagement at all levels, and from various sectors, is critical for ensuring communication strategies and materials are relevant, effective, and well-received. The systematic process followed by this project could benefit future HIV prevention projects. Background: The New HIV Vaccine and Microbicide Advocacy Society teachers training showed that capacity building of teachers in Lagos was an important mechanism to enable students understands HIV prevention. In view of this, NHVMAS structured an annual training to support sustained capacity building programmes for teachers in secondary schools. Methods: NHVMAS undertook a two days training of 23 secondary school teachers in Lagos State using a curriculum ratified by the State Ministry of Education. A 2 item questionnaire pre and post test was administered. Analysis of the pre and post test was conducted so see how participants faired on acquiring new knowledge. Results: There was a significant difference in the pre and post test scores (57.6% vs66.5%; P=0.00). The cohort of trainees had a good understanding of HIV/AIDS &STIs. However very few persons understood the concept of effectiveness: only 9.2% understood about the concept of effectiveness of NPT, and 4.%HIV vaccine respectively. About 76% of participants were aware of the potential of microbicides to prevent HIV infection but only 2.5% of participants understood the mechanisms of action of microbicide in preventing HIV infection. Only 4% understood its potential role for STI and pregnancy prevention. Post training, participants significantly gained knowledge and understanding of these concepts: 38.2% understood about the concept of effectiveness of NPT, and 66.6%HIV vaccine respectively. An average deficient gap of -8.4% and -23.4% was seen in the pre and post test evaluation in answering questions on; the mechanism of action of microbicide and test for safety of microbicide, however, in the second training only -0% and 0% respectively. Conclusions: The study participants seem to have challenges with the understanding of the concept of HIV vaccine research. There may be a need to design a training curriculum that focuses attention on building skills and knowledge about HIV vaccine. 53

154 Posters Posters 02: Community Engagement and Advocacy P02.09 Media Engagement and Reporting on Nigerian - Canadian Collaboration on AIDS Vaccine Research Study in Jos Plateau State Wika Gofwen, Godwin Odemijie 2, Florita Durueke 3, Chuks Okonkwo 3, Morenike Ukpong 3 Plateau Radio Television Corporation, News and Current Affairs, Jos, Nigeria, 2 FRCN, Federal Radio Corporation of Nigeria, News and Current Affairs, Abuja, Nigeria, 3 New HIV Vaccine and Microbicide Advocacy Society, Microbiology, Ibadan, Nigeria Background: The Media is a critical stakeholder in ensuring public education about biomedical HIV prevention technology and can also play roles in setting the research agenda.the Nigerian Canadian Collaboration on AIDS Vaccine (NICCAV) media engagement programme developed model of engaging journalists to ensure it achieves its set objective of building the capacity of the media to effectively educate the general public in Jos Metropolis and other stakeholders on HIV vaccine research and the NICCAV project specifically. Methods: A community based organisation with expertise on community and media engagement in research was engaged on the project to facilitate a systematic engagement of journalists. Specifically, the project through consultation with the Nigeria Union of Journalists (NUJ), identified 30 health reporters from the print and electronic media that were enrolled in the media programme, at the state and national level who equally have their capacity enhanced on (understanding and communicating HIV prevention research) to enable them disseminate culturally and linguistically appropriate information on HIV vaccine research. Media roundtables discussions were held to provide update on the clinical study. Results: The training did show significant increase in knowledge among the journalists, as they identified new knowledge and skills gained. The pre-test and post-test analysis did show significant change in the mean values (p value=0.04). Media roundtables provided update on the clinical study. There was significant increase in the volume of media publications on vaccine research and NPT. Comparative analysis of vaccine and NPT reporting from journalists engaged on the vaccine study was significantly higher than other health reporters that were not enrolled in the project. Conclusions: Investment in a structured media engagement programme on NPT research will facilitate media reporting and advocacy on NPT as well as mobilize community and relevant stakeholders support for vaccine and NPT research. 54 HIV Research for Prevention 204 HIV R4P

155 Posters 03: Correlates of Protection and Exposure Wednesday, 29 October P03.0 HIV-specific T and NK-cell Responses Do Not Correlate with Protection from Sexual Acquisition of HIV Laura Pattacini, Jared M. Baeten 2, Katherine K. Thomas 2, Tayler R. Fluharty, Pamela M. Murnane 2, Deborah Donnell,2, Elizabeth Bukusi 2,3, Allan Ronald 4, Nelly Mugo 2,3, Jairam R. Lingappa 2, Connie Celum 2, Juliana M. McElrath,2, Jennifer M. Lund,2, Partners PrEP Study Team Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 University of Washington, Seattle, WA, United States, 3 Kenya Medical Research Institute, Nairobi, Kenya, 4 University of Manitoba, Winnipeg, MB, Canada P03.02 Type II Interferon Stimulated Genes were Tightly Regulated in Kenyan Commercial Sex Workers who Exhibit Natural Resistance towards HIV Infection. Marcel Gluchowski, Xiaoqiong Yu 2, Renee Douville, Joshua Kimani 3, Frank A. Plummer 4,5, T. Blake Ball 5,6, Ruey-Chyi Su 5 University of Winnipeg, Biology, Winnipeg, MB, Canada, 2 Yangquan Municipal Center for Disease Prevention and Control, Yangquan, China, 3 University of Nairobi, Department of Medical Microbiology, Nairobi, Kenya, 4 Public Health Agency of Canada, National Microbiology Laboratories, Winnipeg, MB, Canada, 5 University of Manitoba, Medical Microbiology & Infectious Diseases, Winnipeg, MB, Canada, 6 Public Health Agency of Canada, National HIV & Retrovirology Laboratories, Winnipeg, MB, Canada Background: The existence of HIV-specific immune responses has been observed in several cohorts of HIV-exposed seronegative (HESN) subjects, yet the ability of these responses to protect against incident HIV infection has not been directly evaluated. Methods: Within a prospective cohort of African male and female HESN who were at high risk of HIV infection as a result of having an HIV infected heterosexual partner, we conducted a nested case-control study to assess HIV-specific immune responses among 76 subjects, including 29 who acquired HIV (cases) and 47 who remained HIV uninfected (controls). Peripheral blood mononuclear cells were collected at the visit prior to HIV acquisition. HIV-specific immune responses were detected by intracellular cytokine staining using global potential T-cell epitope mer peptide pools for Gag and Env, with positive responses defined as dual production of IFN-γ and CD07a for CD8+ T-cell responses, IFN-γ and TNF-α for CD4+ T-cell responses, and IFN-γ and CD07a for NK responses. Logistic regression was used to assess if the frequency and magnitude of immune responses were associated with protection from HIV acquisition. Results: Response rates for CD8+ T-cells were 7.2% and 6.8% in cases and controls, respectively, to Gag,.% and 0.6% to Env (p>0.05). Similarly, no statistically significant differences between cases and controls were observed for CD4+ T-cell responses (Gag: 3.4% vs. 3.4%, Env: 0.0% vs. 2.8% p>0.05) and NK cell responses (Gag: 0.0% vs. 3.4%, Env: 0.0% vs. 6.3%, p>0.05). Furthermore, the magnitude and breadth of responses among responders did not differ between cases and controls. Conclusions: Among HESN with a known HIV positive partner, no relationship was found between T-cell and NK cell responses prior to HIV infection and protection from sexual acquisition of HIV. Our results do not support the hypothesis that natural HIV-specific responses reduce susceptibility to HIV infection in HESN. Background: Interferon stimulated genes (ISGs) are shown to mediate critical anti-viral responses. Cell lines overexpressing ISGs showed reduced viral replication, including HIV. HIV- infection was shown to induce a distinct subset of ISGs that are regulated by interferon regulatory factor- (IRF-). However, low endogenous IRF- level was found to associate with reduced susceptibility to HIV-infection among Kenyan commercial sex worker (CSW). Here, we assessed whether reduced IRF- level, observed in highly HIV-exposed, yet seronegative (HESN) CSW, also correlate with reduced expression and responsive potential of ISG genes. Methods: Endogenous ISG mrna level in peripheral blood mononuclear cells (PBMC) of HESN CSW and control CSW were assessed using quantitative RT-PCR. The responsive potential of these ISGs to type II interferon (IFN) was assessed by stimulating PBMC with IFN-g for 3h; optimized to observe transient IRF- response in HESN and persisting in controls. Results: Not all ISGs are constitutively expressed. Ex vivo HESN PBMC expressed 22 of 67 ISGs examined, while CSW controls expressed 3. Endogenous level of ISGs differed between the 2 groups. ISGs of anti-viral function(irf, CXCL0, TRAILR, ISG56, IFNb, COX2, MXA, TNFa) were significantly lower in the HESN, while the ISGs signaltransducing function(ifit3, MDA5, IFI6) were notably higher in most HESN. Exogenous IFN-g stimulation resulted in significant increases (e.g., CXCL-0, IFN-b, IRF-, IFIT3, STAT-) and decreases (e.g., TRAILR, p6p4, BclXL, BCL2) of some ISGs. Overall, robust increases of CXCL- 0, IRF-7 & IFIT3 were found in HESN with less change in other ISGs. Conclusions: Together, baseline level of ISGs with anti-viral, proinflammatory function was reduced in HESN, with IFN-g responsive ISGs genes being tightly regulated to mount robust, yet transient responses, suggesting an important role for these ISGs in maintaining the HESN phenotype against HIV-infection. 55

156 Posters Posters 03: Correlates of Protection and Exposure P03.03 LB P03.04 LB HIV- of Children with Slow Disease Progression Escapes Autologous Neutralization and Triggers Development of Cross-neutralizing Responses Stefania Dispinseri, Mariangela Cavarelli, Anna Maria Plebani 2, Marianne Jansson 3,4, Gabriella Scarlatti San Raffaele Scientific Institute, Milan, Italy, 2 Foundation IRCCS cà Granda Ospedale Maggiore Policlinico Milan, Milan, Italy, 3 Lund University, Lund, Sweden, 4 Karolinska Institutet, Stockholm, Sweden Background: One-quarter of HIV- infected children has a disease progressing towards AIDS within the first year of age, whereas the majority of them progress slowly over several years. With the purpose to assess the role of the humoral immune response underlying different patterns of disease we studied the kinetics of antibody responses to HIV- and childhood vaccine antigens in slow and rapid progressors followed from birth to 5 years of age. Methods: Autologous and heterologous neutralizing capacity of plasma obtained during follow-up of 25 children was tested with PBMC- and/ or TZMbl-assays, against up to 5 primary viruses for each child or 3 pseudotyped viruses (Tier subtype B and Tier 2 subtype A), respectively. The seroreactivity to HIV- gp4, autologous gp20 V3-loop peptides, and tetanus and diphtheria toxoid was assessed by ELISA. Results: Newborns displayed antibodies towards an immunodominant HIV- gp4 epitope, which were of maternal origin, but did not neutralize the transmitted virus. The child s neutralizing antibodies (Nabs) developed usually within year of age to its own early virus concomitantly with autologous V3-loop directed antibodies in slow, but rarely in rapid progressors. Autologous Nabs persisted throughout disease progression and induced continuous emergence of escape variants. Heterologous Nabs developed within two years independently of disease progression, but their subsequent increase in potency and breadth was a preferential trait of slow progressors. Kinetics of antibody responses to the immunodominant gp4 antigens and childhood vaccine antigens was preserved independently of disease progression. Conclusions: Persistent autologous Nabs triggering viral escape and increase in breadth and potency of heterologous Nabs are exclusive trait of slow progressors. Thus, immune-competence seems to impact the ability to develop potent Nabs, suggesting that early transmitted viruses of slow progressors represent an attractive target for vaccine design, which aims to induce cross-nabs. Modulation of RAS Pathways as a Biomarker of Protection against HIV and as a Means to Improve Vaccine Efficacy Slim Fourati, Monica Vaccari 2, Shari N. Gordon 2, Luca Schifanella 2, Mark Cameron, Brandon F. Keele 3, Xiaoying Shen 4, Georgia D. Tomoras 4, Erik Billings 5, Mangala Rao 5, Amy W. Chung 6, Karen Dowell 7, Chris Bailey-Kellogg 7, Eric Brown 8, Margaret E. Ackerman 8, Namal P.M. Liyanage 2, Diego A. Vargas- Inchaistegui 9, Stephen Whitney 0, Melvin N. Doster 2, Nicolo Binello 2, Poonam Pegu 2, David C. Montefiori, Kathryn Foulds 2, David S. Quinn 3, Mitzi Donaldson 3, Frank Liang 2, Karin Loré 2, Mario Roederer 2, Richard A Koup 2, Adrian McDermott 2, Zhong-Min Ma 4, Christopher J Miller 4, Tran B Phan 5, Donald N. Forthal 5, Matthew Blackburn 2, Francesca Caccuri 2, Guido Ferrari, Devon Thompson 6, Marjorie Robert-Guroff 9, Silvia Ratto-Kim 5, Jerome H. Kim 5, Nelson L. Michael 5, Sanjay Phogat 7, Susan W. Barnett 8, James Tartaglia 7, David Venzon 9, Donald M. Stablein 20, Galit Alter 6, Rafick-Pierre Sekaly 2, Genoveffa Franchini 2 Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL, United States, 2 National Cancer Institute, Animal Models and Vaccine Section, Bethesda, MD, United States, 3 National Cancer Institute, AIDS and Cancer Virus Program, Frederick, MD, United States, 4 Duke Human Vaccine Institute, Durham, NC, United States, 5 U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Silver Spring, MD, United States, 6 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States, 7 Dartmouth College, Computer Science, Hanover, NH, United States, 8 Dartmouth College, Thayer School of Engineering, Hanover, NH, United States, 9 National Cancer Institute, Immune Biology of Retroviral Infection Section, Bethesda, MD, United States, 0 Advanced BioScience Laboratories, Inc., Rockville, MD, United States, Duke University Medical Center, Durham, NC, United States, 2 National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States, 3 Karolinska Institutet, Stockholm, Sweden, 4 University of California, California National Primate Research Center, Davis, CA, United States, 5 University of California, Irvine School of Medicine, Irvine, CA, United States, 6 Advanced BioScience Laboratories, Inc, Rockville, MD, United States, 7 Sanofi Pasteur, Swiftwater, PA, United States, 8 Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United States, 9 National Cancer Institute, Biostatistics and Data Management Section, Bethesda, MD, United States, 20 The EMMES Corporation, Rockville, MD, United States, 2 Case Western Reserve University, Pathology, Cleveland, OH, United States Background: Adjuvants modulate the immune response and can improve the immunogenicity of vaccines. Immunogenicity studies in humans suggest that MF59 is a more efficient adjuvant than alum as it triggers enhanced B and T cell immune responses. Methods: We performed a study (P62MRV44) in macaques using repeated challenges with SIVmac25 powered to benchmark the results of RV44 vaccine that showed limited efficacy in a phase III clinical trial in Thailand. We compared the efficacy of ALVAC-SIV/gp20 vaccine administered with alum or MF59 adjuvants to placebo in the SIV mac25 model. Transcriptional profiling of blood from 54 immunized macaques before vaccination (pre-vax), after immunization with ALVAC-SIV alone (post- st ) and after immunization with ALVAC-SIV combined with the protein/adjuvant MF59 or Alum immunization (post-3 rd ) was performed. Differential expression analysis and pathway enrichment analysis was used to identify genes and pathways associated with protection. A naïve Bayes classifier was build to predict vaccine protection. Results: We found that alum protected macaques from SIVmac25 acquisition (log-rank test: p=0.0205), confirming the main results 56 HIV Research for Prevention 204 HIV R4P

157 Posters 03: Correlates of Protection and Exposure Wednesday, 29 October of the RV44 study. However MF59 did not protect macaques from SIVmac25 acquisition (log-rank test: p=0.562), despite MF59 s ability to elicit higher systemic T-cell and antibodies responses. Association between the changes in transcriptional profiles and risk of SIVmac25 acquisition resulted in the identification of 2-gene expression signature able to predict prior to vaccination protection by ALVAC+alum (ROC: Accuracy=65.2%, p 0.05). Seven of the twelve genes of the signature were related to Ras signaling. Conclusions: System biology revealed that RAS, a signal transducer that facilitates cross talk among B-cells, T-cells and antigen presenting cells, as a biomarker of vaccine efficacy in the ALVAC+alum treated animals. These data suggest that activation of RAS may constitute a novel approach to improve vaccine efficacy against HIV. P03.05 LB Association of HIV- Env Antibody Responses with Postnatal Mother-to-Child Transmission of HIV- via Breastfeeding Genevieve Fouda, Justin Pollara, Erin McGuire, Wesley Rountree, Glenn Overman, Gerald Tegha 2, Deborah Kamwendo 2, Jacob Kumwenda 2, Julie Nelson 3, Larry Liao, Sascha Ellington 4, Caroline King 4, Denise Jamieson 4, Charlie Van der Horst 3, Athena Kourtis 4, Georgia Tomaras, Guido Ferrari, Sallie Permar Duke Human Vaccine Institute, Durham, NC, United States, 2 University of North Carolina Malawi Project, Lilongwe, Malawi, 3 University of North Carolina, Chapel Hill, NC, United States, 4 Centers for Disease Control and Prevention, Atlanta, GA, United States Background: Immune factors in milk may contribute to protect HIVexposed breastfed infants who do not acquire HIV- despite high levels of mucosal virus exposure. To identify humoral immune responses associated with protection against postnatal transmission, we evaluated milk and plasma humoral responses in Malawian HIV-infected women from the control arm of the Breastfeeding, Antiretrovirals and Nutrition study, who did or did not transmit HIV to their infants via breastfeeding. Methods: Transmitting (n=22) and non-transmitting women (n=65) were matched on peripheral CD4 count and postpartum visit. Envspecific IgG and IgA responses were assessed in available milk (n=87) and plasma samples (n=42) using a binding antibody multiplex assay. Antibody dependent cell-mediated cytotoxicity (ADCC) was measured against a postnatally-transmitted HIV- Env variant, and neutralization was measured against a clade C tier virus. Multivariable logistic regression models adjusting for plasma viral load and CD4 count were used to compare responses between groups. Results: The magnitude of IgG responses against the Env antigens, including gp70 B CaseA2 VV2, was similar between transmitters and non-transmitters. However, there was a trend towards an association between lower transmission risk and plasma IgG against gp70 B CaseA2 VV2 with a valine to lysine mutation at position 69 before but not after adjusting for multiple comparisons (OR=0.39, p=0.09, false discover rate (FDR) =0.46). Lower risk of transmission was also associated with high magnitude milk, but not plasma IgA against certain Env antigens, that was not significant after adjusting for multiple comparisons (B con gp40 IgA: OR=0.57, p=0.007, FDR=0.). There was no association between ADCC or tier neutralization responses and transmission. Conclusions: These hypothesis generating results indicate that further studies to examine the role of mucosal IgA and plasma VV2-specific IgG responses in protection against postnatal HIV- transmission may be warranted. 57

158 Posters Posters 04: Correlates of Protection in Highly Exposed Seronegative People P04.0 ABSTRACT WITHDRAWN P04.02 Molecules Involved in the Vitamin-D Pathway Correlate with Higher mrna Expression of Anti-HIV Molecules in HIV Exposed Seronegative Individuals Wbeimar Aguilar-Jiménez, Manuel G. Feria, Eliuth D. Arcia, Wildeman Zapata,2, Maria T. Rugeles Grupo Inmunovirología. Universidad de Antioquia UdeA, Medellin, Colombia, 2 Grupo Infettare. Universidad Cooperativa de Colombia, Medellin, Colombia Background: Vitamin D (VitD) is an endogenous immunomodulator that could protect from HIV- infection. We recently found that high levels of VitD and its receptor are associated with natural resistance to HIV- infection; up-regulation of the anti-inflammatory cytokine IL-0, and the induction of defensins in mucosa might be part of the mechanisms involved in this association. However, several other molecules might be involved in this protection. Methods: To further explore this issue, a case-control study using samples of 58 HIV--exposed but seronegative (HESN) individuals, and 59 non-exposed healthy controls (HCs) was carried out. mrna relative units (RU) of APOBEC3G, ELAFIN, TRIM5alfa, SAMHD and Catelicidin in peripheral blood mononuclear cells (PBMCs), oral and genital mucosa were quantified by qrt-pcr. Circulating VitD and mrna levels of VDR were previously reported and used for correlations. Results: HESNs had significantly higher mrna RU of the antiviral molecules APOBEC3G and ELAFIN in PBMCs and SAMHD and Catelicidin in oral mucosa compared to HCs. Positive correlation between VDR and ELAFIN mrna in PBMCs of HESNs was found. Conclusions: These results suggest that high levels of APOBEC3G, ELAFIN, SAMHD, and Catelicidin are associated with natural resistance to HIV- infection. VitD may be up-regulating the expression of ELAFIN as observed for other anti-hiv- molecules. However, further studies are required to define causal associations. 58 HIV Research for Prevention 204 HIV R4P

159 Posters 05: Family Planning P05.0 Wednesday, 29 October P05.02 Contraceptive Use and Pregnancy Incidence among VOICE Participants in Uganda Carolyne A. Akello, Katherine E. Bunge 2, Clemensia Nakabiito, Brenda G. Mirembe, Mary G. Fowler,3, Anupam Mishra 4, Zvavahera Mike Chirenje 5, Jeanne Marrazzo 4, Connie Celum 4, Jennifer E. Balkus 4,6 Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 Magee Women s Research Institute, Pittsburgh, PA, United States, 3 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 4 University of Washington, Seattle, WA, United States, 5 University of Zimbabwe, Harare, Zimbabwe, 6 Fred Hutchinson Cancer Research Center, Seattle, WA, United States Stop and Save Children from HIV! Rural-urban Differentials, Barriers and Motivators of Longterm Contraception in Zimbabwe, Mixed Methods Results Munyaradzi Kenneth Dodzo,2, Munyaradzi Paul Mapingure, Noah Taruberekera 3 Population Services International Zimbabwe, Research and Metrics, Harare, Zimbabwe, 2 University of Zimbabwe, Population Studies, Harare, Zimbabwe, 3 Population Services International, Research and Metrics, Johannesburg, South Africa Background: Recent HIV prevention trials have required use of effective contraceptive methods to fulfill eligibility for enrolment. We compared pregnancy rates in participants enrolled in the VOICE (MTN-003) trial who initiated depot medroxyprogesterone acetate (DMPA) or combined oral contraceptives (COCs) at enrollment (new users) relative to those already on DMPA or COCs. Methods: Data were analyzed from HIV- seronegative participants enrolled in Uganda. Prior to enrolment, information on contraceptive type and initiation date was obtained; contraception was provided to new users. Participants received urine pregnancy tests at monthly follow-up visits. Cox proportional hazards models stratified by baseline contraceptive method were used to compare pregnancy incidence among new users (initiated 60 days prior to enrollment) and established users (initiated >60 days prior to enrolment). Participants were censored after the first observed pregnancy on study. Results: Of 322 women enrolled, 296 were COC or DMPA users; 82 (28%) were new users (50 [6%] initiated DMPA and 32[39%] initiated COCs) and 24 (72%) were established users. Overall pregnancy incidence was 3.35 per 00 person-years [p-yrs] (49/367 p-yrs), with an incidence in DMPA users of 5.39 per 00 p-yrs (3/24 p-yrs) compared to per 00 p-yrs (36/26 p-yrs) in COC users. In new DMPA users, pregnancy incidence was 0.20 per 00 p-yrs versus 3.48 per 00 p-yrs in established DMPA users (adjusted hazard ratio [ahr] = 2.84; 95% confidence interval [CI] ). Similarly, in new COC users, pregnancy incidence was per 00 p-yrs versus per 00 p-yrs in established COC users (ahr 2.08; 95% CI ). Conclusions: New contraceptive users in VOICE had an increased pregnancy risk. Pregnancy incidence was high among both new and established COC users. New DMPA and all COC users participating in HIV prevention trials may benefit from intensive contraceptive counseling and provision of less user-dependent methods. Background: Socio-economic and biologic vulnerabilities predispose women to HIV infection, leading to higher maternal risk and infection of babies. Contraception lowers HIV incidence, viral load, pregnancy and vertical transmission. Long-acting reversible contraceptives (LARCs) - implants and IUDs - help overcome access barriers, common in poverty, emergency or abuse cases. In Zimbabwe, their prevalence is 2% but CPR is 60%. PSI seeks their full potential by integrating them in HIV prevention, sexual and reproductive health. We aim to identify barriers, facilitators and correlates of LARCs by place of residence. Methods: We used two studies. One involved 85 women (5-49 years) in a national, multi-stage household survey, using UNIANOVA in SPSS to analyse data. Correlates were built from multiple items scored to 5. Qualitative data were analysed thematically from 44 in-depth interviews with adult men and women. 24 women were using LARCs and 2 were not. 4 men had partners using LARCs and 4 were not. Results: Urban women, 6%, were more likely to use LARCs than rural, 3%, p< Dual use of LARCs and condoms was higher among urban, 5%, than rural women, 3%, p< Partner support was higher among urban women, 24%, than rural, 8%, p< Scaled availability was better among urban (3.92) than rural (3.69), p< 0.00 as was social support - urban (2.8) vs. rural (2.62), p< 0.00 and self-efficacy - urban (3.07) vs. rural (2.56), p< Qualitative data showed that LARCs spell peace of mind, low maintenance costs, flexibility and freedom from parenting duties. But they interfere with menses, link with infertility and threaten womanhood. Fear of pain, infection and side effects are other barriers. Insertion or removal skills portray them as elitist, costly and inaccessible. Conclusions: Prevalence of LARCs is low and myths abound. PSI must promote awareness and knowledge about LARCs. Program must create opportunity, improve ability and increase motivation, especially among rural women. 59

160 Posters Posters 05: Family Planning P05.03 Capacity Strengthening and Training of Government Nurses on Long-acting Reversible Contraceptive (LARC) Methods in Kigali, Rwanda P05.04 Strategies for Successful Promotion and Implementation of Contraceptive Method Mix in MTN-020/ASPIRE: The Why and How at Kampala Site Rosine Ingabire, Etienne Karita, Nurilign Ahmed, Roger Bayingana, Julien M. Nyombayire, Robertine Sinabamenye, Jean N. Nduwamungu, Honoree Uwamahoro, Sarah Strunk, Amanda Tichacek,2, Susan Allen,2 Projet San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 2 Emory University, School of Medicine, Department of Pathology and Laboratory Medicine, Atlanta, GA, United States Background: In Rwanda, adult women 5-49 have higher HIV prevalence rate (4%) compared to men (2%). Each year, nearly half (47%) of all pregnancies in the country are unintended. Preventing unintended pregnancies among HIV positive women through family planning (FP) is the lowest cost perinatal HIV prevention strategy and contraceptive use is associated with longer survival in HIV positive women. HIV negative women enrolled in clinical trials should also avoid pregnancy. Methods: Project San Francisco (PSF) provided training in Long Acting Reversible Contraceptives (LARC), the copper IUD and Jadelle implant in 6 Kigali government clinics to enhance access to and uptake of the most effective family planning methods. From June May 204, PSF conducted monitoring & evaluation of LARC services in Kigali Rwanda. The overall numbers of nurses trained, provision of LARC methods at the government clinics, and LARC provision were measured. Results: PSF conducted 9 LARC training sessions with 59 nurses trained in IUD/Implant insertions. From , these nurses provided 5,28 LARC methods (996 IUDs and 4,285 Implants). During this period, the percentage of family planning clients who requested LARC methods increased annually in the 6 target clinics: IUD provision among FP clients increased from % in 2009 to.2% in 200, 5% in 20 and 9% in 202. Implant insertions increased from 8.6% in 2009 to 7.% in 200 to 2% in 20 and 37% in 202. PSF continues to provide technical assistance for the provision of LARC methods and between June 202- March 204, 2,937 (3,009 IUDs and 9,928 Implants) have been provided. Conclusions: When nurses are comfortable and competent with inserting LARC methods, they are more likely to successfully promote and provide these methods to both HIV positive and negative clients. In turn, these clients are very likely to opt for LARC when they are available. Established LARC services in government clinics enhance prong 2 of PMTCT and readiness for prevention trials. Betty Kamira, Clemensia Nakabiito, Justine Kamya Nsangi, Joselyne Nabisere, Christine Nagawa, Samuel Kabwigu, Fred Kapaata, Flavia K. Matovu MU-JHU Research Collaboration, Kampala, Uganda Background: To enroll in ASPIRE; participants must be on an effective method of contraception ranging from COC, DMP, Implants, IUCD and sterilization. In Uganda, there is a low contraceptive prevalence and a high unmet need (43%) of use. This was reflected in the VOICE study where uptake for Long Acting Reversible Contraceptive (LARC) was only 6%. In an attempt to enhance Contraceptive Method Mix (CMM) in ASPIRE, the Kampala team describes how this was made successful. Methods: Preparatory activities started prior to site activation with formation of the Contraceptive Action Team (CAT) under the leadership of the MTN Contraceptive Steering Committee with the aim of exploring misconceptions, identifying barriers, and increasing LARC uptake. Following this, 2 members were declared contraceptive experts, and charged with spearheading CMM. These liaised with existing f/planning clinics to train study counselors on counseling messages and clinicians in inserting and removal of LARCs. We started linkages with the community and satellite clinics to demystify myths and misconceptions using flip charts and models and the community found this exciting. All methods were made available onsite, assigned a nurse for logistics and created a room for providing the LARC. Results: There is an increase in the use of IUCDs and implants and a decrease in injectables and pill use after the inception of CAT. Previously, contraception was limited to COCs and DMPA. LARCs (Implants, IUCD) and tubal ligation were being offered at the nearby Hospital. Now all methods are available on-site except tubal ligation. Conclusions: Method Mix and expansion of LARC methods is possible in a low resource limited setting. For success, information is key, ongoing refresher training, regular evaluations, peer support, good political will ongoing participant care and counseling are all important. 60 HIV Research for Prevention 204 HIV R4P

161 Posters 05: Family Planning P05.05 Wednesday, 29 October P05.06 Contraceptive Preference among HIV High Risk Women Pre-screened for a Phase III Vaginal Microbicide Trial in South-Western Uganda Sylvia Kusemererwa, Andrew Abaasa 2, Emanuel Aling 2, Margaret Kalibbala 2, Sarah Nakato 2, Juliet Kyomugisha 2, Neliette Van Niekerk 3, Annalene Nel 3, Anatoli Kamali 2 Medical Research Council/ Uganda Virus Research Institute, Uganda Research Unit on AIDS, Entebbe, Uganda, 2 MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 3 International Partnership for Microbicides, Paarl, South Africa Knowledge and Use of Modern Family Planning Methods in Fishing Communities along Lake Victoria, Uganda Annet Nanvubya, Juliet Mpendo, Ali Ssetaala, Julius Ssempiira, Annet Nalutaaya, Mathias Wambuzi, Steven Asiimwe 2, Leslie Nielsen 3, Fredrick Makumbi 4, Noah Kiwanuka,4 UVRI-IAVI HIV Vaccine Program, Entebbe, Uganda, 2 Kabwohe Clinical Research Center (KCRC), Kabwohe, Uganda, 3 International AIDS Vaccine Initiative, New York, NY, United States, 4 Makerere University College of Health Sciences, School of Public Health, Kampala, Uganda Background: Women s contraceptive choices vary according to the type of relationships they have and other aspects of their life. We describe the preference of modern contraceptive methods among HIV high risk women screened to participate in a Phase III vaginal microbicide trial. Methods: Medical Research Council/Uganda Virus Research Institute, Uganda Research Unit on AIDS, in collaboration with International Partnership for Microbicides (IPM), is evaluating safety and efficacy of a dapivirine vaginal ring among healthy HIV negative women in a multicentre Phase III trial (The Ring Study). One of the study eligibility criteria is being on a stable form of contraception in order to reduce the pregnancy rate. As part of the screening process, women are identified from townships and fishing communities within 50 km from the centre. Data on use of modern contraception methods (Pills, Injectables, Implants, IUCDs, Surgical) are collected, and confirmed by a contraceptive card or presence of an implant or IUCD strings. Women who are willing are provided by the research centre with contraception of choice (pills, injectables) or referred to a provider (Implants, IUCDs, Surgical). The proportion of women per method used was determined. Results: A total of 489 women, with a mean age of 28 years, were pre-screened (age range 8 to 44 years). Most (306, 63%) were already using contraception: 55% were on Injectables, followed by Implants (28%), IUCDs (8%), Pills (7%) and only 2% had undergone a Surgical method. Of the women who were not on any method, 06 (58%) accepted Injectables, 40 (22%) preferred Implants, 33 (8%) oral Pills and 4 (2%) chose IUCDs. Conclusions: There is high contraceptive use among HIV high risk women and the majority prefer Injectables. This is encouraging and will hopefully reduce discontinuation of volunteers from the trial due to pregnancy. Background: Interventions geared towards reduction of unwanted pregnancies and elimination of mother to child transmission of HIV have led to the integration of family planning (FP) and HIV-care services into the general health care package. Fishing communities are another key population for HIV infection in Uganda with incidence and prevalence ~3-6 times higher than estimated national rates. These communities are being targeted for HIV prevention trials where FP is a requirement. There is paucity of data on the knowledge and use of modern FP including hormonal, barrier and permanent methods in fishing communities. Methods: Data are accrued from a baseline survey of 29 HIVuninfected individuals aged 8-49 years enrolled between September 20 and March 203 from 8 fishing communities. At the 2-months post baseline visit, data on knowledge and use of family planning were collected through a semi structured questionnaire. Results: About three quarters (688/29, 77%) of the volunteers provided data on FP. Knowledge about family planning was high, 87% (477/688). Just above a third, 35% (595/688) reported current use of at least one modern FP method. Use of modern FP by a respondent and/or their sexual partner was more frequently reported by females than males (4.6% vs. 29.2%, p< 0.00). Factors associated with higher use of modern FP were: increasing level of education (p=0.02), being currently married (p< 0.00) and currently having other sexual partner/s (p< 0.00). Reasons for non-use of modern FP methods were: desire for more children (30.6 %), currently not being in a sexual relationship (27%), fear of side effects (2.2%), currently pregnant/breast feeding (7.8%) being young (7.%) and partner refusal (5.2%). Conclusions: Knowledge of modern FP methods among fishing communities along Lake Victoria, Uganda is high but use is still low. High fertility desires, not being in a sexual relationship and fear of perceived side effects are key barriers to use of modern FP. 6

162 Posters Posters 05: Family Planning P05.07 P05.08 Introduction of Long Acting Reversible Contraception (LARC) in a Clinical Trial Setting in Kwazulu-Natal Arendevi Pather, Jayajothi Moodley, Naureen Cele, Vermala Juggernath, Gita Ramjee, MTN 020 Medical Research Council of South Africa, HIV Prevention Research Unit, Durban, South Africa Background: Although debatable, recent studies have shown a possible increased risk of HIV acquisition with injectable hormonal contraception. In KwaZulu-Natal (KZN), the epicenter of the HIV epidemic, injectable contraception is widely used. Based on the recommendation by the World Health Organization (WHO) to increase contraceptive options, the Microbicide Trials Network (MTN) formed a Contraceptive Action Team (CAT), tasked to improve the contraceptive options at MTN research sites. As part of this initiative, the Medical Research Council (MRC) Clinical Research Sites (CRSs) embarked on a drive to provide Intrauterine Devices (IUDs) and implants at the CRSs. Methods: Challenges included lack of LARC knowledge; myths and misconceptions about LARC, participant fears about partner objection and LARC side effects; preference to remain on current method, provision of training to clinical staff and accessing LARC. Strategies used included: community education and awareness conducted during community meetings and outreach campaigns; ongoing training provided to clinical staff on LARC insertions in collaboration with the KZN Department of Health (DoH); accessing of LARC from private suppliers and current negotiation with DoH; counselling and education on LARC during waiting room discussions at the CRSs and individual contraception counselling sessions. At these discussions, participant fears about partner objection and fear of side effects of LARC were also addressed. Results: We have received positive feedback from the wider community regarding LARC. Community programmes ensure that myths and misconceptions about LARC are dispelled. Clinical staff have been trained on insertion of LARC at three CRSs. Conclusions: The CAT initiative has been successful in introducing and implementing LARC within the MRC CRSs in KZN. Despite this, ongoing efforts are still required to increase the number of clinical staff trained on IUD and implant insertions at the CRSs and to increase the utilisation of LARC. Combination Prevention for PMTCT Prongs & 2: HIV Testing for Couples and Long Acting Reversible Family Planning/dual Method Use Promotion in Zambia Bellington Vwalika,2, Inambao Mubiana 3, William Kilembe, N Munir, Kathleen Wu, N Ahmed, Ashey Appiagey, B Getachew 4, R Parker 4, J Seed 5, Danielle Dang 6, Amanda Tichacek 5, Kristin Wall 5, Susan Allen 5 Zambia Emory HIV Research Project, Lusaka, Zambia, 2 University of Zambia, Lusaka, Zambia, 3 Zambia Emory HIV Research Project, Ndola, Zambia, 4 Zambia Rwanda HIV Research Group, Atlanta, GA, United States, 5 Rwanda Zambia HIV Research Group, Atlanta, GA, United States, 6 Rwanda Zambia HIV Research Group, Lusaka, Zambia Background: Prongs and 2 of PMTCT focus on prevention of heterosexual HIV and prevention of unplanned pregnancy in HIV+ women but receive little attention in the HIV prevention world. Couples HIV counseling and testing (CVCT) reduces new heterosexual infections. Long acting reversible contraceptives (LARC), in particular the IUD and the implant, are effective modern methods that are not available to most African women with or at risk of HIV. The UK Department for International Development has funded a program to integrate CVCT, family planning promotion, and LARC for clinic visitors and couples in the community with the goal of averting3,900 new HIV infections. Methods: Formative research with providers; clinic clients in family planning, infant vaccination, and ART clinics; and couples seeking CVCT informed development of training and promotional materials for nurses, counselors, and community promoters. A program dovetailing simultaneous ramp-up of supply and demand was implemented in 50 clinics in 6 Zambian cities in Lusaka, Southern and Copperbelt Provinces. Results: 99 nurses from government clinics were LARC-trained and 306 CVCT counselors, community health workers and LARC clients/peer educators were trained in couples family planning counseling (CFPC) and promotion. From March 203 to March 204, 37,28 couples received CVCT and 4,564 of those also received CFPC. In this same time frame, 269 IUD and 6977 Jadelle were inserted, and 357 IUD and 906 Jadelle were removed after a mean of 2.6 years of use. Dual method use (barrier + LARC) was emphasized with discordant couples. Conclusions: Combining HIV testing with family planning and targeting couples has the potential to prevent many new heterosexual and perinatal infections. Both LARC training and LARC methods are sustained once administered. With a concerted effort and coordination of supply and demand, the IUD and implant can be integrated in government clinics and HIV programs. The combination is feasible, cost effective and sustainable. 62 HIV Research for Prevention 204 HIV R4P

163 Posters 06: Financial Incentives P06.0 Economic Incentives to Reduce HIV Risks among Male Sex Workers: A Randomized Pilot in Mexico Wednesday, 29 October P06.02 Unanticipated Impact of Financial Incentives on HIV Patients and Providers: Findings from a Qualitative Substudy (HPTN 065) Omar Galarraga, Sandra G. Sosa-Rubi 2, Carlos J. Conde-Glez 3, Luis Juarez-Figueroa 4, Andrea Gonzalez 5, Florentino Badial- Hernandez 5, Sergio Bautista-Arredondo 2, Caroline Kuo 6, Don Operario 6, Kenneth H. Mayer 7 Brown University, Health Services Policy & Practice, Providence, RI, United States, 2 National Institute of Public Health, Health Economics, Cuernavaca, Mexico, 3 National Institute of Public Health, Infectious Disease Research Center, Cuernavaca, Mexico, 4 National Institute of Public Health, Mexico City, Mexico, 5 Clinica Condesa, Mexico City, Mexico, 6 Brown University, Behavioral & Social Science, Providence, RI, United States, 7 Fenway Health & Harvard University, Global Health, Boston, MA, United States Background: We test economic incentives for HIV risk reduction among male sex workers (MSW). N=265 participants were randomized into 4 groups. Methods: In Treatment, they received a medium conditional incentive ($50 USD/each time) only if they were free of new, curable STI at months 6 and 2. In Treatment 2, they received a high incentive ($75 USD/ each time) if they were free of new, curable STI at months 6 and 2. Controls did not receive incentive even if free of STI. In Unconditional Incentive, they received medium incentive ($50 USD at months 6 and 2) regardless of STI status. All groups received inconvenience fees ($0 USD/each time at baseline, month 6 and month 2). All incentives were in the form of food and grocery vouchers. We recruited from Alameda Central and Clínica Condesa (largest HIV clinic in Mexico City> 9,000 patients). All eligible men took part in a standard HIV education session after completing baseline measures and before randomization. All received HIV/STI testing; those infected were offered treatment (for curable STI). Results: At baseline, mean age was 25, 76% were single, and 35% completed high school. On average they had 6 sexual partners per week, and 83% had used condoms with last three clients. Most were street-based (70%), but worked at bars/clubs also (30%). Most had at least one STI (63%): 9% had active syphilis, % had chlamydia; and HIV prevalence was 38%. The average median transaction price per sexual act was USD$25 with 40% higher price for unprotected sex. Conclusions: Results provide strong rationale for structural intervention for MSW. High HIV rates and higher payments for unprotected sex justify economic intervention which has been demonstrated to be feasible and acceptable. Preliminary efficacy on STI outcomes, condom use, number of partners, and transaction prices will be presented at proposed session. Theresa Gamble, Jill Stanton, Elizabeth Greene, Jamilah Taylor, Allison P. Pack, Victoria Shelus, Elizabeth E. Tolley, Sheldon T. Brown 2, Wafaa El-Sadr 3,4, for the HPTN 065 Study Team FHI 360, Durham, NC, United States, 2 James J. Peters VA Medical Center, Bronx, NC, United States, 3 Columbia University Mailman School of Public Health, ICAP, New York, NY, United States, 4 Harlem Hospital, New York, NY, United States Background: The HPTN 065 (TLC-Plus) study evaluated the effect of providing quarterly $70 financial incentives (FI) to HIV infected patients on antiretroviral therapy (ART) with HIV RNA< 400 copies/ml to encourage adherence. Nineteen HIV care sites randomized to the FI intervention dispensed 39,359 FI gift cards over two years in the Bronx, NY and Washington, DC. Methods: We conducted semi-structured interviews with 75 patients from 4 sites and 2 site investigators (SIs) (mostly clinicians); three focus groups were conducted with 2 staff members from 0 sites. Data were analyzed to determine the effect of the FI on aspects other than adherence. Results: While the purpose of the FI was to influence ART adherence, patients and staff reported more engagement in care (improved visit adherence, patient/provider relationships, and general preventive care) and increased opportunities for HIV and general health education. Patients reported that the FI increased their ability to manage their HIV, improved their attitudes towards the disease and its treatment, and caused other behavior change (e.g. motivation not to drink so as to remember to take their ART). Many patients also reported that the FI met a real financial need. During the program, patients felt cared for by staff, and staff felt good about doing something positive for their patients. Conversely, for some patients, the gift cards invoked a sense of entitlement (i.e. patients felt they deserved, rather than earned, the gift card), reminded them of their HIV status, and may have devalued their efforts to remain adherent. Especially at the beginning of the intervention, some clinics experienced changes in clinic flow and had difficulty managing the influx of patients. Conclusions: The use of FI intended to influence ART adherence can affect both patients and providers in unanticipated ways. While some effects were, at least transiently, negative, most of the additional findings were positive, with the potential to improve HIV care and overall patient health. 63

164 Posters Posters 06: Financial Incentives P06.03 The Impact of Implementing a Financial Incentive Program for Viral Suppression on the Clinic Environment: A Qualitative Substudy of HPTN 065 Elizabeth Greene, Theresa Gamble, Elizabeth E. Tolley, Allison P. Pack, Jill Stanton, Jamilah Taylor, Victoria Shelus, Jason Leider 2, Wafaa El-Sadr 3,4, Bernard Branson 5, for the HPTN 065 Study Team FHI 360, Durham, NC, United States, 2 Jacobi Medical Center, New York, NY, United States, 3 Columbia University Mailman School of Public Health, ICAP, New York, NY, United States, 4 Harlem Hospital, New York, NY, United States, 5 Centers for Disease Control and Prevention, Atlanta, GA, United States P06.04 Understanding of Viral Load among Participants Receiving Financial Incentives for ART Adherence: Findings from a Qualitative Substudy of HPTN 065 Elizabeth Greene, Jamilah Taylor, Allison P. Pack, Jill Stanton, Victoria Shelus, Elizabeth E. Tolley, Lawrence J. D Angelo 2, Wafaa El-Sadr 3,4, Theresa Gamble, for the HPTN 065 Study Team FHI 360, Durham, NC, United States, 2 Children s National Medical Center, Washington, DC, United States, 3 Columbia University Mailman School of Public Health, ICAP, New York, NY, United States, 4 Harlem Hospital, New York, NY, United States Background: HPTN 065 (TLC-Plus) studied the effect of providing quarterly $70 financial incentives (FI) to HIV-infected patients on antiretroviral therapy (ART) with HIV RNA< 400 copies/ml to encourage ART adherence. Nineteen HIV care sites randomized to the FI intervention dispensed a total of 39,359 FI gift cards over two years in the Bronx, NY and Washington, DC. Methods: Semi-structured interviews were conducted with 75 patients from 4 sites and 2 site investigators (SIs) (mostly providers); three focus groups were conducted with 2 staff members from 0 sites. Data were analyzed qualitatively to determine the impact of the FI program on clinic environment. Results: The majority of patients interviewed noticed no effect on their clinic experience due to the FI program. SIs and staff reported that the study-imposed timing of quarterly viral load tests necessary to qualify for FIs often conflicted with typical clinical care schedules and caused clinics to become busier as patients visited frequently or adhered to appointments. SIs saw a benefit to this (better engagement in care), but it also increased demands on staff time and space and interfered with clinic flow. Some SIs and staff found determining eligibility, tracking and disbursing gift cards to be logistically challenging. Some staff also had difficulty dealing with patients who felt entitled to FIs and discussing eligibility for FIs in semi-private spaces. SIs and staff described a learning curve as processes were developed to improve logistics, clinic flow, and FI and patient tracking, and as patients and providers became educated about program requirements. Most SIs attributed smooth implementation to well-trained and dedicated staff and provider buy-in. Many indicated that positive patient interactions with the FIs outweighed implementation challenges. Conclusions: A large scale FI program posed challenges for providers and staff that were mitigated by effective systems and education without negative effects on patients clinic experiences. Background: HPTN 065 (TLC-Plus) evaluated the effect of providing quarterly $70 financial incentives (FI) to HIV-infected patients on antiretroviral therapy (ART) to encourage ART adherence. Viral load (VL) suppression, defined as HIV RNA< 400 copies/ml, was used as a marker for ART adherence. Nineteen HIV care sites were randomized to the 2-year FI intervention in the Bronx, NY and Washington, DC. Methods: We conducted semi-structured interviews with 75 patients ages 4-72 from 4 sites and 2 site investigators (SIs) (mostly clinicians). Qualitative data were analyzed to determine perceptions about VL. Clinical data were collected to determine patients VL suppression status. Results: Based on their descriptions, patients understanding of the meaning of VL or their specific VL results was variable; almost half misunderstood VL. Many confused it with CD4 count, some thought VL should ideally be high, and some admitted lack of understanding despite efforts from providers. Several used the term undetectable incorrectly, suggesting they had heard this term but did not grasp its meaning. More patients recognized that ART adherence would result in a good VL, even though some were unaware of the definition of a good value. While some patients said their understanding of VL improved during the study, over half said that it did not. In contrast, most SIs thought that many of their patients understood their VL results, and several felt the study increased understanding of VL. There was no association between observed viral suppression and accurate understanding of VL or its relationship to ART adherence. Conclusions: Many patients did not accurately understand VL even though it was the outcome that qualified them to receive the FI. Clinicians thought patients understood VL when often they did not. However, accurate understanding of VL was not always necessary to comprehend that VL improved with ART adherence, nor was it related to viral suppression among patients interviewed. 64 HIV Research for Prevention 204 HIV R4P

165 Wednesday, 29 October Posters 07: Good Participatory Practices and Community Involvement P07.0 P07.02 Ethics of HIV Prevention in Research Literacy Obiajulu A. Amuamuziam, Morenike O. Ukpong, Florita C. Durueke, Oluwatosin B. Alaka, Olayide Akanni 2,3 New HIV Vaccine and Microbicide Advocacy Society (NHVMAS), Programme, Lagos, Nigeria, 2 New HIV Vaccine and Microbicide Advocacy Society (NHVMAS), Administrative, Lagos, Nigeria, 3 Journalists Against AIDS (JAAIDS) Nigeria, Programme/Administrative, Lagos, Nigeria Background: When NHVMAS studied the methodology section of the 2007 and 200 IBBSS survey reports, it was clear that though the methodology was sound, there was a gap in the ethics of the conduct of trials as there was no evidence of community engagement in the design of the survey. Members of the communities engaged with these surveys are becoming concerned with their continued participation in surveys that do not translate to interventions for them. While NHVMAS makes effort at increasing awareness of the public, researchers and policy makers to this clause of the HIV research policy, it also wants to focus on building the capacity of the community. Methods: NHVMAS undertook three days training for gatekeepers (IDU, FSW and MSM) communities in seven sites where the IBBSS 204 survey was to take place. The training focused on empowering the community gatekeepers as trainers on what they need to know about research, its importance, informed consent, confidentiality and so on and how they can also transfer same to their peers. Participants learnt about the new and existing HIV prevention tools such as microbicides, HIV vaccines, medical male circumcision and female condoms. An interactive approach was engaged. Analysis of the pre and post test was conducted to compare knowledge of participants on topics treated before and after training. Results: There was difference in the pre and post test (45.4vs 56.2; p=.07626e-05). One of the impact of the training was the demand made by community members during the planning of the IBBSS 204 survey for a pre-planning consultation with the community of MSM before conclusion on study design. There have been other reports of community members demanding for informed consent and ethics clearance prior to conduct of research with community members. Conclusions: When community gatekeepers become more research literate, they can become actively engaged with research design. Also empowered to make demands for changes in research processes that could address their community needs. Implementing Good Participatory Practice (GPP) in HVTN505 - The HIV Vaccine Trials Network (HVTN) Experience Gail B. Broder,2, James P. Maynard,3, Shelly T. Karuna,4, Chuka Anude 5, Magda E. Sobieszczyk 6, Scott M. Hammer 6, HVTN 505 Protocol Team of the NIAID-funded HIV Vaccine Trials Network Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 HIV Vaccine Trials Network, Community Engagement Unit, Seattle, WA, United States, 3 HIV Vaccine Trials Network, Communications and Community Engagement Units, Seattle, WA, United States, 4 HIV Vaccine Trials Network, Clinical Development Unit, Seattle, WA, United States, 5 DAIDS/NIAID/NIH, Vaccine Clinical Research Program, Bethesda, MD, United States, 6 Columbia University College of Physicians and Surgeons, Division of Infectious Diseases, Department of Medicine, New York, NY, United States Background: HVTN has valued community involvement in its operations since it began, and continued this practice with HVTN505, a Phase 2B trial in MSM and transwomen. During HVTN505 implementation, results from the iprex trial and subsequent FDA approval of Truvada for preexposure prophylaxis (PrEP) led the HVTN to consider changes to HIV prevention services offered to participants (ppts). Several community consultations informed our actions that addressed the GPP principles of transparency and the standard of HIV prevention. Methods: Community members were part of the protocol team from the outset, contributing to study design, informed consent materials, engagement and recruitment efforts, and communications planning. Post-iPrEx, HVTN505 ppts were surveyed about their intent to use PrEP. Consultations were held with Global Community Advisory Board members representing all network sites and with a wider group of community stakeholders to discuss the iprex results and standards of HIV prevention. Community input was sought on the advisability of providing PrEP for trial ppts interested in using it, and on the mechanisms of that provision. Post-FDA approval, a second consultation was held regarding implementation of such a plan. Results: Stakeholders felt strongly that ppts should be educated about PrEP and counseled about how to access it. The study was amended and incorporated the potential for 20% uptake in PrEP use among ppts. As discussions in the field continue about the changing standard of HIV prevention, HVTN is working to make Truvada available free of charge to interested HVTN505 ppts. This evolving discussion will continue to inform future trial designs. Conclusions: The HVTN embraces the GPP guidelines and continues its efforts to fully implement them. Involving the community at all levels is key in the HVTN s structure and all of its trials. HVTN505 provided a unique opportunity to engage the community at a time when the conversation about HIV prevention was evolving particularly rapidly. 65

166 Posters Posters 07: Good Participatory Practices and Community Involvement P07.03 P07.04 Increasing the National Impact and Uptake of Good Participatory Practice Guidelines for Biomedical HIV Prevention (GPP): Three Country Case Studies Stacey Hannah, Julius Ecuru 2, Udom Likhitwonnawut 3, Catherine Slack 4, Ntando Yola 5 AVAC, New York, NY, United States, 2 Uganda National Council of Science and Technology, Kampala, Uganda, 3 Thai NGO Coalition on AIDS, Chiang Mai, Thailand, 4 HIV AIDS Vaccines Ethics Group, Pietermaritzburg, South Africa, 5 Desmond Tutu HIV Foundation, Cape Town, South Africa Background: The Good Participatory Practice Guidelines (GPP) provide a normative framework for stakeholder engagement in HIV prevention trials. GPP has been applied at site, network and sponsor levels to enhance trial-specific and general site operations. GPP contains recommendations, not requirements, and no entity was established to ensure implementers fulfillment of practices. There is increasing interest in incorporating GPP into national trial oversight processes as one method of strengthening positive impact. We review case studies from three key countries for HIV prevention research. Methods: The Uganda National Council of Science and Technology (UNCST) oversees regulatory approval of research nationally. In 20, UNCST initiated incorporation of GPP into revision of national ethics guidelines for clinical research. In South Africa, recommendations for national GPP implementation were developed through a series of consultations with research staff and stakeholders. Efforts are also ongoing to consider select high-impact activities for Research Ethics Committees reviewing engagement plans in protocols. In Thailand, implementation of GPP at research centers led to formation of a national CAB, development of a national GPP plan within the Ministry of Public Health and incorporation of GPP into ethics review. Results: Adoption of minimum requirements for GPP in clinical research has helped establish the guidelines as standard practice in several countries and work toward greater impact of these efforts. Cases described provide models for adaptation and use in additional countries engaged in prevention research. Conclusions: GPP was developed to enhance the ethical and effective implementation of HIV prevention research. National adoption and other strategies to increase impact are critical to realizing this aspirational goal. The field may benefit by replication of these models in additional countries. Innovation in Stakeholder Engagement: Piloting a Monitoring and Evaluation Toolkit Stacey Hannah, Stephanie Seidel 2, Sinazo Pato 3, Monique Oliff 4 AVAC, New York, NY, United States, 2 Global Alliance for Tuberculosis Research, New York, NY, United States, 3 Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 4 Independent Consultant, Mombasa, Kenya Background: The Good Participatory Practice Guidelines (GPP) for HIV prevention trials were adapted for TB drug trials and are being used generically in additional settings. As practices expand, increasing focus rests on GPP impact. A cross-field, cross-disease monitoring and evaluation (M&E) toolkit was developed; it has been piloted widely, using a variety of methodologies, research settings, and respondents. Methods: Pilot sites were identified both from centers conducting novel combination TB drug trials and those conducting biomedical HIV prevention trials, all with significant history of engagement programs. Example pilot sites included the Aurum Institute and FACTS Consortium, both in South Africa, and the Kenya Medical Research Institute in Kilifi, Kenya. Program staff piloted a series of mapping and data collection tools using quantitative and qualitative methods to survey targeted clinical and engagement activities through the trial lifecycle. A web-based interface for data analysis was also tested. Tools were assessed for effectiveness in documenting and analyzing engagement activities aimed to improve the research process and stakeholder outcomes. Results: Indicators of success, relevant data points and data collection methodology were refined. Respondents helped revise key parameters of engagement work, including community advisory board activities, small group/individual consultation, communications and outreach to stakeholders beyond trial community. The pilot helped refine links between engagement and clinical outcomes such as participant adherence. Conclusions: Incorporation of feedback from sites with leading GPP experience led to development of an effective, relevant, user-friendly toolkit for monitoring and evaluation of community and stakeholder engagement activities. Wide use of the toolkit is needed to build an evidence base and to better understand the impact of this work on research and stakeholder-related outcomes. 66 HIV Research for Prevention 204 HIV R4P

167 Wednesday, 29 October Posters 07: Good Participatory Practices and Community Involvement P07.05 P07.06 Implementing Community Involvement in National Institutes of Health (NIH) HIV/AIDS Clinical Trials Networks Rona Siskind, Neetha S. Morar 2, Russell D. Campbell 3, Jeffrey Schouten 3 Division of AIDS, Workforce Operations, Communications and Reporting Branch, Bethesda, MD, United States, 2 South African Medical Research Council, HIV Prevention Research Unit, Westville, South Africa, 3 Fred Hutchinson Cancer Research Center, Office of HIV/AIDS Network Coordination, Seattle, WA, United States Background: Researchers and funders in HIV clinical prevention and therapeutic research have long recognized the value of including community stakeholders in research planning and implementation to ensure its integrity, relevance and acceptance. Early on, NIH-funded HIV/ AIDS clinical research has required community engagement. In 2009 the Recommendations for Community Involvement in HIV/AIDS Clinical Trials Research were developed to help guide these efforts. With similar principles, the Recommendations complement the Good Participatory Practice (GPP) guidelines (developed by UNAIDS/AVAC 2007, updated 20); both are used across the NIH HIV/AIDS Clinical Trials Networks. Methods: NIH, the Office of HIV/AIDS Network Coordination (HANC) and communities from across the NIH HIV/AIDS Clinical Trials Networks worked to develop these Recommendations. Focusing on the community advisory board (CAB) model, the Recommendations delineate roles/ responsibilities of staff and community throughout each stage of the research process, from initiation through results dissemination and site closure, taking the social and cultural context into consideration. Results: The NIH HIV/AIDS Clinical Trials Networks and sites use the Recommendations, GPP and other local guidelines to develop CABs or equivalent groups to engage global and local communities. This allows for community input into the scientific agenda, study design, eligibility, informed consent and recruitment materials. Dissemination of research results continue to be improved to reach the broader community. Better training on the Recommendations and GPP is needed and is currently underway. Conclusions: The NIH Clinical Trials Networks and sites have successfully engaged community using the Recommendations and GPP guidelines, adapting them as needed, to be responsive to community suggestions and concerns. Both encourage the use of multiple engagement strategies to develop community partnerships and more comprehensive approaches to community engagement. Research Participants Skills Development as Peer Educators in their Communities Neetha Shagan Morar South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: Peer education is a global community-based HIV prevention intervention to improve health. In 2005, researchers initiated a peer education programme, nested within HIV prevention clinical trials. This paper describes the influence of this programme on skills development of trial participants, as they exercised agency in their role as peer educators. Methods: Qualitative methods using round table discussions and semi structured interviews were used to collect data from 22 women on their experiences as peer educators. From June to November 2009, data was collected, audio-recorded and transcribed by trained interviewers. Content thematic analysis was used to identify themes which included peer educators experiences, opinions of training and skills development. Results: Experience and training within the peer education programme advanced skills development of the peer educators. Using the newly acquired information on HIV prevention, sex and health promotion, they provided health-related advice and knowledge to members of their family and community. The experience improved their self-esteem and confidence as they developed communication skills to discuss HIV, sex and research procedures with their partners and family members. The peer programme was empowering and improved their knowledge of HIV and research. Conclusions: Peer experiences contributed to skills development where peer educators were able to conduct outreach and education sessions. It enhanced their agency as they developed confidence to engage their family and community on HIV prevention and research related issues. Training and partnering with clinical trial participants as peer educators is an effective and sustainable community based approach that develops their skills and enhances agency among women. 67

168 Posters Posters 07: Good Participatory Practices and Community Involvement P07.07 P07.08 Challenges for Community Engagement in HIV Research in Thailand Niwat Suwanphatthana Thai NGO Coalition on AIDS, Muang, Thailand Background: Thailand has been involved in several breakthroughs in HIV research for many years including RV44, iprex and HPTN 052. These are multisite trials involving general population or specific populations. Community Advisory Board (CAB) is the main mechanism for community engagement of these trials. Most CABs have limited autonomy in providing recommendations due to power relationship between researchers and CABs. Research institutions handled CAB formation with limited inputs from communities. CAB inputs are reduced to reviewing inform consent documents. Most institutes don t have structured plan to strengthen CAB capacity. CABs are not educated about their role and responsibility resulting in confusion and conflict of interest. Methods: Promotion of meaningful community engagement in HIV prevention The Thai NGO Coalition on AIDS (TNCA) with supports from AVAC is promoting community engagement including:. Good Participatory Practices (GPP) promotion for research institutions and stakeholder 2. GPP integration in national policy and mechanism Results:. Stakeholder consultations on the 2 nd edition of GPP 2. GPP trainings of trainers for research institutions and a lessons learned workshop 3. The Thai National AIDS Management Center (NAMc) and the Department ofdisease Control with inputs from TNCA organized series of workshops on HIV research and strategic programing involving key stakeholders and targeted populations. 4. The Thai National AIDS Committee established the Sub-committee of Biomedical HIV Prevention recommended by NAMc. 5. TNCA formed a National CAB consisted of members of existing CABs in the country to enhance CAB capacity in HIV and clinical research. Conclusions:. GPP is integrated in the national AIDS plan for biomedical HIV research. 2. Ethical committees and research institutes become familiar with GPP. Some research institutes develop stakeholder engagement plans based on GPP. Skills Training of Kenyan Health Care Providers Attending to Men who Have Sex with Men Improved Services Two Years Post Training Elise Maria van der Elst, Bernadette Kombo, Evans Gichuru, Anisa Omar 2, Helgar Musyoki 3, Susan Marie Graham 4, Adrian D. Smith 5, Don Operario 6, Eduard Joachim Sanders 7 Kenya Medical Research Institute, HIV/AIDS Social Science, Kilifi, Kenya, 2 Kilifi County, Ministry of Health, Kilifi, Kenya, 3 National AIDS and STI Control Programme, National MARPs Programme, Nairobi, Kenya, 4 Washington University, Infectious Disease Medicine, Seattle, WA, United States, 5 Oxford University, Department of Public Health, Oxford, United Kingdom, 6 Brown University, Public Health, Providence, RI, United States, 7 Kenya Medical Research Institute, Epidemiology, Kilifi, Kenya Background: Men who have sex with men (MSM) are often at high risk for HIV acquisition, and access to and quality of health services within this population are negatively affected by hetero-normative attitudes. A recently developed online MSM-healthcare education program www. marps-africa.org helped reduce homoprejudice among healthcare providers (HCP) in Kenya. In this study we used qualitative methods to explore the provision of MSM healthcare services two years posttraining. Methods: We held 8 focus group discussions (FGD) with 54 participants overall; 3 included HCP, 2 included district AIDS-coordinators, and 3 included local LGBT-members. Participants discussed availability, acceptability and accessibility of HIV-healthcare for MSM. HCP also discussed changes in health service practices after completing the training. FGD were recorded, transcribed verbatim, and analyzed using the grounded theory approach. Results: HCPs described improvements in their service provision to MSM-patients post-training. They felt more empowered and comfortable asking male patients about same-sex behavior, which contributed to nonstigmatizing HIV-service environments for MSM. Four unique dimensions for improving MSM utilization of healthcare were identified: ) HCP expressed a clear need for pre-service MSM sensitivity training, including the need for national guidelines to manage sexually transmitted anal infections (STI); 2) AIDS-coordinators advocated for reporting tools specifically developed for MSM in terms of program implementation and capacity building; 3) MSM described a need for programs to reduce stigma, especially among male HCPs; 4) All felt that the government should endorse improved health-skillstraining for MSM patients. Conclusions: Positive impacts of this skills training were reflected in HCP attitudes two years post-intervention. Scaling-up of efforts will rely on continued policies to include MSM in healthcare, programs to reduce stigma in health settings, and directives guiding MSM-STI-service delivery. 68 HIV Research for Prevention 204 HIV R4P

169 Posters 08: HIV Care P08.0 Wednesday, 29 October P08.02 Choosing Appropriate Interventions and Defining Locally Achievable Standard of Care for HIV Prevention Trials in Africa: A Multidisciplinary Approach Susan Allen, William Kilembe 2, Mubiana Inambao 3, Bellington Vwalika 4, Shabir Lakhi 4, Kristin Wall 5, Amanda Tichacek, Gordon Streeb 6 Rwanda Zambia HIV Research Group, Emory University, Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA, United States, 2 Rwanda Zambia HIV Research Group, Emory University, Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Ndola, Zambia, 4 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Lusaka, Zambia, 5 Emory University School of Public Health, Epidemiology, Atlanta, GA, United States, 6 Emory University, Economics and Political Science, Atlanta, GA, United States Background: Per capita incomes (PPP) in most African countries are < $2000/year and per capita expenditures on health < $00. The cost per HIV infection averted (HIA) should be less than per capita income. These metrics should inform what interventions are appropriate to test and what local standards of care (SOC) should include. To reduce AEs and unplanned pregnancy, trials should also provide other health services. Methods: Phase I-III trials conducted at the Rwanda Zambia HIV Research Group (RZHRG) were assessed to identify common preventable infectious diseases reported as AEs, and pregnancies. Separately, a non-rzhrg study protocol for a large phase III trail of ART treatmentas-prevention (TasP) was reviewed by a team with 25+ years of HIV, development, and ethics research in Africa. SOC services were evaluated and ethical concerns were identified. Results: In 5 RZHRG trials, malaria, helminth infections, and diarrhea were common: in one trial, 6/24 HIV- participants were diagnosed with urinary schistosomiasis. Bed nets, chlorine, hand soap, and helminth treatment could prevent most of these AEs. Contraception (FP) was provided and there were no pregancies. In the TasP trial, the majority of participants will have a spouse. Voluntary HIV Testing and Counseling (VTC) is a trial procedure and Couples VCT (CVCT) should also be, as recommended by WHO. The protocol refers to family testing but this is not a prevention strategy. CVCT is associated with a reduction in new HIV infections and should be explicitly included in protocols and procedures. HIA for TasP should be compared with HIA CVCT, MC, and FP. Conclusions: Both trials and participants would benefit from low-cost screening and treatment services for endemic diseases such as bed nets, routine deworming, soap and chlorine as well as provision of contraceptives. TasP trials should analyze costs and have a rationale for testing interventions with HIA > PPP. Excluding locally affordable HIV prevention services including CVCT from trial SOC is unethical. The Continuum of HIV Care in Peru - Where Are we Now? Key Lessons from an Estimation in the Context of Very Limited Data Carlos F. Caceres, Kelika Konda 2, Alfonso Silva-Santisteban, Ximena Salazar, Lottie Romero, Segundo Leon, Jeffrey D. Klausner 3 Cayetano Heredia University, Unit of Health, Sexuality and Human Development, Lima, Peru, 2 University of California at Los Angeles, Program in Global Health, Lima, Peru, 3 University of California at Los Angeles, School of Medicine, Los Angeles, CA, United States Background: The role of universal access to treatment in combination HIV prevention has been established. Estimation of coverage across steps in a continuum of care cascade (CCC), a key approach to monitor progress and identify barriers, is novel in Peru and most of Latin America. We aimed to build an estimation of the CCC for Peru, identify potential barriers defining losses across the process, and establish data gaps. Methods: We drew from data from both peer-reviewed publications and official estimates (i.e. Global Fund Evaluation Reports, Government and UNAIDS bulletins) to estimate coverage for the CCC steps in Peru. We assessed whether a single CCC was appropriate, as opposed to specific CCC by sub-populations. Then we used the most plausible parameters to generate estimates of the cascade steps. We provide recommendations for initial policy changes and further research. Results: Of 75,000 adults estimated to be living with HIV in Peru in 203, 22,000 were women. Tested primarily in connection with reproductive health services, they generally showed a better CCC profile than men, with earlier access to and higher retention in care, and more frequent viral suppression after 6 months. Among men, data are available primarily for MSM and male-to-female transwomen (TW). Key barriers across the CCC include: low HIV testing rates (only 27% of HIV+ know their status) and late registration in care (as reflected in CD4 counts around 00 at ARVT initiation). Only 8% were virally supressed. Conclusions: The CCC for women in Peru shows progress towards universal access, reflecting success of programs to prevent mother to child transmission, and possibly better health seeking. For MSM/TW, active promotion and facilitation of biannual HIV testing is needed to improve serostatus awareness, linked with easier access to care, with strategies to avoid losses to follow up prior to ARVT initiation. Data for other men are extremely limited. An integrated, reliable information system to monitor progress is urgently needed. 69

170 Posters Posters 08: HIV Care P08.03 HIV Care Continuum among MSM in Latin America Using Online Sexual Networking: Is Engagement in Care Related to Sexual Risktaking? Jessica F. Magidson, Katie B. Biello 2,3, Steven A. Safren,2, Joshua G. Rosenberger 4, David S. Novak 5, Kenneth H. Mayer 2,6, Matthew J. Mimiaga,2,3 Massachusetts General Hospital / Harvard Medical School, Psychiatry, Boston, MA, United States, 2 The Fenway Institute, Boston, MA, United States, 3 Harvard School of Public Health, Boston, MA, United States, 4 George Mason University, Department of Global and Community Health, Fairfax, VA, United States, 5 OLB Research Institute, Online Buddies, Inc., Cambridge, MA, United States, 6 Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, United States Background: The HIV/AIDS epidemic in Latin America is concentrated among men who have sex with men (MSM). Yet, it has been difficult to characterize the HIV care continuum in this population, as many may not self-identify as MSM. Further, given recent evidence of reduced likelihood of sexual HIV transmission in the context of viral suppression, examining whether HIV-infected individuals not in care are engaging in HIV transmission risk behavior is important for secondary prevention. Methods: The current study recruited via an online sexual networking website to reach men who may not self-identify as MSM but are seeking male sex partners online--a population at high risk for HIV acquisition and transmission. Primary aims were to examine ) the HIV care continuum in this sample; and 2) whether HIV-positive individuals not in care had higher rates of unprotected anal intercourse (UAI) than those in care at each step of the care continuum. Surveyed 29,787 active members of an MSM sexual networking site in Latin America on HIV testing and HIV diagnosis, receipt of medical care and ART for HIV treatment, ART adherence, and UAI in the past three months. Percentages were calculated and logistic regressions were performed. Results: Overall, 74.% reported ever being tested for HIV and 9.0% reported HIV diagnosis. Of the HIV-positive individuals, 20.0% reported not being in medical care, 29.% not receiving ART, and 55.3% reported suboptimal adherence to ART. HIV-positive individuals not in care reported greater UAI compared to those in care (OR=.40; 95%CI=.0-.78). Those not ART adherent reported greater UAI than those who were adherent (OR=2.02; 95%CI= ). Conclusions: Findings estimate the HIV care continuum among a large sample of MSM in Latin America using the internet to meet sex partners. A key limitation of this study was not having a measure of viral load. Despite this, findings still suggest that secondary prevention for HIV-positive MSM in Latin America not in care or ART adherent are warranted. 70 HIV Research for Prevention 204 HIV R4P

171 Posters 09: HIV Testing and Counseling P09.0 Wednesday, 29 October P09.02 Impact of South-to-South Technical Assistance from Rwanda & Zambia in Couples Voluntary HIV Counseling and Testing (CVCT) Achievements in 9 Countries VCT Acceptance and High Level of HIV Stigma and Discriminatory Attitudes towards HIV Infected People among Male Prisoners in Northern Thailand Nurilign Ahmed, Ashley Appiagyei 2, Annie Mwaanga 2, Cortney Robinson 2, Robertine Sinabamenye, Elias Gudo 2, Helman Banda 2, Muyunda Mulenga 3, Bella Mutumwinka, Bella Siangonya 3, William Kilembe 4, Mubiana Inambao 3, Etienne Karita, Susan Allen 5 Project San Francisco, Kigali, Rwanda, 2 Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Zambia Emory HIV Research Project, Ndola, Zambia, 4 Zambia Emory Research Project, Lusaka, Zambia, 5 Emory University, School of Medicine, Department of Pathology and Laboratory Medicine, Atlanta, GA, United States Background: CVCT is recommended by WHO for HIV prevention and is associated with reduction in HIV, STI and unplanned pregnancy. Fewer than 5% of African couples have been jointly tested and it is thus critical that CVCT be expanded. The RZHRG Center of Excellence provides training and technical assistance (TTA) to countries and partners wishing to implement CVCT as standard of care in ongoing service delivery or research activities. Methods: Primary data from RZHRG was analyzed to describe TTA events from TA events were classified as high-level advocacy (including policy, workshops and study tours of RZHRG sites), technical assistance for needs assessments, data management or other program implementation needs and training (of trainers, service providers, community-level promoters and others). Data from recipient countries was analyzed to investigate resulting country-adapted CVCT models, as well as changes in uptake of CVCT. Results: RZHRG conducted 6 TA events in 9 countries ranging from development of national strategic plans and international workshops to CVCT curriculum adaptations and trainings. Countries such as Botswana and Uganda have demonstrated higher success in CVCT implementation through the roll out of a national CVCT strategy. Increased uptake of CVCT led to increased identification of discordant couples. Countries such Ghana and Ivory Coast have made strong commitments to expand CVCT services and are making good progress to that end. Through RZHRG TA, several countries including Namibia and South Africa have an extensive number of trainers across levels and sectors available to scale-up CVCT training efforts. Kenya, Mozambique, Tanzania and Swaziland have sent high level observers to Rwanda and Zambia. Conclusions: Through TTA, recipient countries were able to acquire tools and skills and multiple levels, which enabled them to more effectively plan for and implement CVCT. This program increased capacity of countries to prioritize and expand locally tailored CVCT activities. Suwat Chariyalertsak,2, Chonlisa Chariyalertsak 3, Kriengkrai Yotruean 3, Chutima Charuwat 3, Kriengkrai Srithanaviboonchai Research Institute for Health Sciences Chiang Mai University, Chiang Mai, Thailand, 2 Faculty of Medicine Chiang Mai University, Community Medicine, Chiang Mai, Thailand, 3 Chiang Mai Public Health Office, Chiang Mai, Thailand Background: Although prisoners should be a venue for providing HIV testing, few studies have explored the factors that may influence HIV VCT acceptance especially on HIV stigma and discriminatory attitudes related to PLHIV among prisoners. Methods: This study aims to measure HIV-related stigma and discrimination among male prisoners in one prison in Chiang Mai, Northern Thailand. Only male ethnic minority prisoners were invited to participate by using self-administered questionnaires for those who can read and write Thai and by face-to-face interview for prisoners who have no education. The questionnaire developed by the Global Stigma and Discrimination Indicator Working Group aims to measure three domains of this phenomenon included fear of HIV transmission, manifestations of HIV-related stigma, and discriminatory attitudes towards PLHIV. Participants who responded affirmatively to any question within any one domain were considered to possess HIV stigma believe and discriminatory attitudes. The data were also disaggregated by age, education, incarcerated period, type of prosecution and HIV VCT uptake. Results: There were 282 prisoners with average age of 3 years old. The domain receiving the highest affirmative responses was manifestations of HIV-related stigma (87.6%), followed by discriminatory attitudes towards PLHIV (52.8%), and fear of HIV transmission (48.2%). The subjects who never received an HIV testing compared to ever had HIV VCT (RR.34, CI= ) and had no more than primary education compared to had higher education (RR.34, CI=.05-.7) revealed more fear of HIV contraction. The less educated prisoners also responded poorly discriminatory attitudes (RR.56, CI= ) compared to the higher educated group. Other factors and incarcerated period were not associated with affirmative responses in either domain. Conclusions: Stigma-reduction program in prison are urgently needed to gain HIV VCT and reduce their risk behavior of HIV infection during and after discharged from the facility. 7

172 Posters Posters 09: HIV Testing and Counseling P09.03 HIV Test: Which Is your Best? An Italian Survey on Testing Preferences among MSM Giulio Maria Corbelli,2, Sandro Mattioli, Stefano Pieralli, Michele Degli Esposti, Roberto Cascioli, Valerie Taccarelli Plus Onlus, Bologna, Italy, 2 European AIDS Treatment Group, Bruxelles, Belgium Background: HIV testing opportunities in Italy are frequently limited to the hospital setting. Other strategies could improve HIV testing uptake, especially in targeted populations. Methods: We aimed at better understanding about testing preferences in the Italian MSM population. An internet-based survey was conducted between March 0 and April 3, 204. The survey was promoted on Plus onlus social networks and gay websites. Results: 348 questionnaires were collected. Responders were 88% male, aged (35%) or (25%), homosexual (8%) or bisexual (9%). Half or them had an HIV test within 2 years (56%) 8% never tested for HIV. 6% had more than 2 sexual partners in the past year. Reported condom use in the past year was: always 39%, always but once %, sometimes 27%, never 4% (0% had no penetrative sexual intercourse). Most known places to have an HIV test are hospital (95%), STI clinic (58%) and chemical analysis laboratory (54%); most used: hospital (73%), STI clinic (30%), laboratory (22%); 5 responders reported having had a self-test at home. Preferred places is self-testing at home (53%), hospital (36%), pharmacy (32%) and headquarter of an organization (3%). Most known testing method is draw blood from vein (97%), which is also most used (80%) but the least preferred (3%) while saliva (65%) and finger prick (56%) are the preferred choices. Physicians are the preferred operator (54%) followed by self-testing (46%), nurses (46%) and peer-volunteers (39%). The ideal HIV test should be: reliable (86%), with no medical prescription (75%), free (63%), rapid (55%), with no personal information collected (45%), with the opportunity to speak with a peer-counselor (36%). Conclusions: Awaiting for the results and bureaucratic obligations represent the major barriers to HIV test in Italy. Home-testing and community-based testing seem to be among the best ways to offer new opportunities for HIV test though home testing will not allow any kind of support for newly diagnosed people. P09.04 Test for Triage: A New Approach to Community-based HIV Testing and Counselling Rachel Baggaley, Kathryn Curran, Cheryl Johnson, Anita Sands 2, Martina Brostrom 3, Vladanka Andreeva 4 WHO, World Health Organization, Department of HIV/AIDS, Geneva, Switzerland, 2 WHO, World Health Organization, Essential Medicines and Health Products, Geneva, Switzerland, 3 UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva, Switzerland, 4 UNAIDS, the Joint United Nations Programme on HIV/AIDS, Bangkok, Thailand Background: Despite recent achievements in scaling-up HIV testing and counselling (HTC), two major challenges remain: ) increasing the number of people who know their HIV status, particularly populations at highest risk for HIV, and 2) ensuring correct HIV test results are delivered. WHO recommends community-based HTC (CBHTC) in generalized epidemics and for key populations in all epidemics. However, in some settings, there are regulatory barriers and lack of support for use of HIV rapid diagnostic test (RDTs) at point-of-care by community providers. There are also concerns that quality of testing and accuracy of results may be sub-optimal in many settings. Methods: A test for triage approach represents a new, simple way to support CBHTC. Community providers conduct a single RDT. Individuals with a reactive test result are immediately linked to a facility for further HIV testing (starting at the beginning of the national testing algorithm) and assessment for treatment. Individuals with a non-reactive test result are given their results, referred for appropriate HIV prevention services and recommended for re-testing according to national guidelines. Results: This new approach encompasses both () expansion of CBHTC performed using a single RDT and (2) emphasis of immediate linkage for HIV diagnosis and treatment assessment at health facilities. The simplification of CBHTC to a single RDT followed by linkage of people who have a reactive test result to further testing with a new specimen and a second reader at a facility could reduce operator errors, thus, leading to more reliable HIV test results, especially for low prevalence populations (Figure ). Supportive HTC policies, clear messages for clients and providers to understand test results and strong linkage-tocare are critical to the success of this approach. Conclusions: This simplified approach to CBHTC may address current challenges in HTC. It could be implemented along with efforts to strengthen quality assurance. 72 HIV Research for Prevention 204 HIV R4P

173 Posters 09: HIV Testing and Counseling P09.05 Wednesday, 29 October P09.06 Opportunities for HIV Prevention among Couples in Durban, South Africa William Kilembe, Mammekwa Mokgoro 2, Annie Mwaanga, Miriam Kamusoko 2, Tarylee Reddy 3, Elisabeth Dissen, Jonathan Davitte 4,5, Shumba Phiri Hilda, Mark Brockman 6, Thumbi Ndung u 2, Susan Allen 5,7 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 2 HIV Pathogenesis Program, Durban, South Africa, 3 Medical Research Council of South Africa, Durban, South Africa, 4 Rwanda Zambia HIV Research Group, Ndola, Zambia, 5 Emory University, Atlanta, GA, United States, 6 Simon Fraser University, Burnaby, BC, Canada, 7 Rwanda Zambia HIV Research Group, Atlanta, GA, United States Background: Couples Voluntary Counseling and Testing (CVCT) reduces HIV transmission by up to 60% among discordant couples but uptake is low in a priority setting. Rwanda Zambia HIV Research Group (RZHRG) in collaboration with the HIV Pathogenesis Programme (HPP) at the University of KwaZulu-Natal (UKZN) conducted a pilot project to expand of CVCT and to assess opportunities to implement HIV prevention strategies that target couples at local clinics in Durban, South Africa. Methods: In February 203, trainers from RZHRG conducted CVCT training and supervision for counselors and health promoters at five clinics around Durban. Client level indicators including age, gender, pregnancy status, ART status, and sero status of both partners, were collected for all couples as a component of routine CVCT service operation. Descriptive analyses are presented here. Results: A total of 20 counselors and 28 promoters completed training. Out of 908 couples (86 individuals) that underwent CVCT, 50% of men and 62% of women had been previously tested alone; only 4% had been tested as a couple. Prevalence of HIV was 42% and prevalence of discordant couples was 29% (9% F+M-, 0%F-M+). Almost half of the couples were within the age group 20 to 29 years. This group had the highest prevalence of HIV and contributed the highest proportion of discordant couples. Only 2% of discordant couples had the positive partner on ARVs (3% M-F+, 8 % M+F-) and 30% of the concordant positive couples with one or both partners on ARVs. In 95 couples (0%), the woman was pregnant. Of these, 4 (5%) were discordant couples where the woman was the negative partner and had never been tested as couples before; only in one of these couples was the man on ARVs. Conclusions: The burden of HIV in Durban is high. CVCT, an intervention that targets the largest risk group in sub-saharan Africa, would greatly benefit the couples in Durban as an HIV prevention strategy and as an entry point to care and treatment services aimed at reducing HIV incidence. Effectiveness and Impact of Promotion of Health Rights by Sex Workers for Reducing HIV Prevalence in the Eastern of Democratic Republic of Congo Mambo Amisi Modeste,2 Humanitarian Action for Health and Community Development, Public Health, Bukavu, Democratic Republic of the Congo, 2 Institut National de Sante Public, Epidemiologie, Kinshasa, Democratic Republic of the Congo Background: Since 994, the Eastern Democratic Republic of Congo has been a theater of armed conflict, violence, and serious human rights violations. Two out of three women have experienced the violence personally. While the whole community suffers, sex workers and MSM transgender people are particular victims of discrimination, rape and sexual violence. The 20 annual report of the National Program for fighting against HIV/ AIDS, indicated that a 4.7% HIV prevalence among sex workers in Eastern Democratic Republic of Congo. This information drove the sex workers gathered at the AHUSADEC NGO to initiate a program in 202, Club mutual pleasure of sex workers, with the goal of promoting prevention and sharing information about STIs and HIV by sex workers to sex workers, the community and customers, and to promote human rights and advocacy in the towns of Bukavu and Goma in the Eastern Democratic Republic of Congo. Methods: We re organized into six solidarity communities. 276 received training in peer education on HIV/AIDS prevention. Through our voluntary counseling and testing center, we promote education, training and sensitization for HIV testing. In the conflict zone we educated and trained an army group and women about HIV prevention. Results: In sex workers were sensitized about human rights and trained as peer educators in STI-HIV/AIDS prevention; - 308,600 male and,803 female condoms used by 27 army group member in the conflict zone; - 04 sessions conducted by sex workers peer educators on IST-VIH/ SIDA and human right with 27 army groups in Eastern of D.R. Congo; soldiers from army groups know their HIV status through UNHCR and VCT of AHUSADEC; lubricants distributed; - 74 soldiers from army groups who are HIV + received ARV treatment through the UN mission HIV section; were identified as having sex with condoms in February, 793 in July and 3705 in November 203, an increase of 75%. Conclusions: The effective involvement of sex workers in the promotion of human health could reduce the prevalence of HIV / AIDS to 2% in sentinel areas (Walikale, Masisi, Rutshuru and Minova) in territory occupied. The very low economic power of sex workers is our challenge. 73

174 Posters Posters 09: HIV Testing and Counseling P09.07 Engaging Young Adult Clients of Retail Pharmacies for HIV- Testing in Coastal Kenya Peter M. Mugo, Henrieke Prins, Elizabeth Wahome, Grace Mwashigadi, Alexander Thiong o, Evanson Gichuru, Anisa Omar 2, Susan M. Graham,3, Eduard J. Sanders,4 Kenya Medical Research Institute, Kilifi, Kenya, 2 Ministry of Health, Kilifi, Kenya, 3 University of Washington, Seattle, WA, United States, 4 Oxford University, Headington, United Kingdom P09.08 Correlates of Voluntary HIV Testing and Collection of Test Results among Male Clients of FSWs in Three States of India Karikalan Nagarajan, Sheela Godbole, Sucheta Deshpande 2, Ramesh Paranjape National AIDS Research Institute, Pune, India, 2 Public Health Foundation of India, New Delhi, India Background: Adults in resource-limited countries frequently use retail pharmacies as the first or only source of treatment for various ailments. The aim of this study was to assess whether young adult clients of retail pharmacies can be referred for HIV- testing and engaged for HIV- prevention research. Methods: We requested five pharmacies to refer clients meeting predefined criteria (8-29 years of age and requesting treatment for fever, diarrhoea, sexually transmitted infection (STI) symptoms or body pains) to selected study health facilities, where HIV- testing and screening for an ongoing HIV- prevention study was offered. Using multivariable logistic regression, we determined client characteristics associated with uptake of HIV- testing. Results: From February through July 203,,490 pharmacy clients met criteria for referral (range of weekly average by pharmacy: 4-35); 377 (25%) reported for screening at a health facility, 353 (24%) were HIV- tested and 27 (9%) met criteria for the prevention study. Of those tested 4 (3.9%) were HIV- infected. Test uptake varied significantly by referring pharmacy, and was higher for clients who presented at the pharmacy without a prescription vs. those with a prescription, and for clients who sought care for fever or STI symptoms vs. those who sought care for body pains. Conclusions: About a quarter of retail pharmacy clients engaged for HIV- prevention research were tested for HIV-. Clients seeking care directly at the pharmacy (i.e., without a prescription) and those with fever or STI symptoms were more likely to take up HIV- testing. Engagement of adult pharmacy clients for HIV- testing may identify undiagnosed individuals and offers opportunities for HIV- prevention research. Background: Male clients of female sex workers (FSWs) are a high risk (HRG) and bridging population driving the heterosexual HIV epidemic in India. National AIDS control programme has prioritized scaling up of voluntary HIV testing (VT) among HRG s. Impact of VT, dependent upon collection of test results (CR), is of significance in this bridging population. We assessed the correlates of VT&CR among male clients in India to understand its determinants. Methods: Data were drawn from the cross-sectional IBBA survey of male clients of FSWs between in three high prevalence states of Tamil Nadu, Andhra Pradesh and Maharashtra, India.Informed consent was obtained. Multivariate logistic regression models were used to assess the correlates of VT&CR and to identify factors preventing clients from collecting test results Results: Of 4803 clients, 78(6.2%) reported VT, of whom 70 (90.9%) of clients had collected reports (CR). Clients exposed to STI advertisements [AOR.9(CI ), p< 0.05] and Key clinics (branded STI clinics) [AOR.2(CI ), p< 0.05 respectively] were more likely to take-up VT&CR. Higher education status(>2 grade) increased the likelihood of VT&CR uptake [AOR.8(CI ), p< 0.05], while unorganized laborers and truck/transport workers were less likely to undergo VT&CR [AOR 0.6(CI ), p< 0.05 AOR 0.7(CI ) p< 0.0 respectively]. Higher education was linked with increased likelihood of collecting results: grades 6-2 and grades > 2 were less likely to have not-collected test reports [AOR 0.4(CI ), p< 0.05 AOR 0.(CI ), p< 0.05 respectively]. Conclusions: We highlight the overall low uptake of VT among clients of FSW and that not all test- seekers are collecting reports. Program focus to improve test-seeking as well as report collection is warranted through IEC campaigns as they have positively influenced uptake of VT&CR. Specific interventions among less educated, unorganized laborers and truckers would help in improving VT and CR among clients. 74 HIV Research for Prevention 204 HIV R4P

175 Posters 09: HIV Testing and Counseling P09.09 Wednesday, 29 October P09.0 Socio-demographic Factors Associated with Uptake of HIV Counseling and Testing (HCT) among Nigerian Youth Ayodeji Oginni, Otibho Obianwu 2, Sylvia Adebajo 2 Population Council, Abuja, Nigeria, 2 Population Council, HIV, Abuja, Nigeria Background: HCT is an important gateway for HIV prevention interventions as it educates sero-negative individuals on HIV preventive behaviours and enables seropositive individuals to gain access to treatment, care and support services. We evaluated the sociodemographic factors associated with HCT-uptake among Nigerian youth aged Methods: Secondary data analysis was conducted on Nigeria s 202 National HIV/AIDS & Reproductive Health Survey data. Multivariable logbinomial regression analysis was used to estimate adjusted prevalence ratio (APR) with 95% Confidence intervals. Results: Of the 0,09 youth, half were aged 5-9, 66.9% were single, 65.7% were rural dwellers, 20.7% had no education, 46% were students & 3.% were employed. About 0.5% ever had HCT & 3.5% tested positive in the survey. Multivariable analysis revealed that the aged [APR=.67(.4-.96)] were more likely to have had HCT than the aged 5-9. HCT-uptake increased with educational level [primary- APR=2.29( ); secondary APR=3.48( ) & higher- APR=6.68( )]. The non-catholic [APR=.60( )] and the Catholics [APR=.85( )] Christians were more likely to have had HCT than the Muslims. Those having comprehensive knowledge of HIV [APR=2.09( )] were twice more likely to have had HCT. Students [APR=0.80( )] were less likely to have had HCT than the employed. Those from poor-households [APR=0.63( )] were less likely to have had HCT than those from average-households. Conclusions: HCT-uptake among young Nigerians is very low despite the increased availability of free HCT services in the country. The fact that being employed and having higher educational level and household wealth are associated with HCT-uptake suggests that socioeconomic barriers to HCT-uptake persist among young people. The association with age may be due to age of consent barriers faced by adolescents. More youth-friendly interventions aimed at increasing HCTuptake among young Nigerians are urgently needed. HCT as a Point of Entry for HIV Prevention and Treatment - Profile of Men and Women Presenting at HCT Centers in Durban, South Africa Jessica Lyn Phillip, Renee Street, Susie Hoffman 2, Kelly Blanchard 3, Theresa Exner 4, Elizabeth Kelvin 5, Gita Ramjee,6, Joanne Mantell 2 HIV Prevention Research Unit, Medical Research Council, Durban, South Africa, 2 HIV Center for Clinical and Behavioral Studies and Columbia University, New York State Psychiatric Institute, New York, NY, United States, 3 Ibis Reproductive Health, Cambridge MA, USA and Johannesburg, South Africa, Johannesburg, United Kingdom, 4 HIV Center for Clinical and Behavioral Studies, New York State Psychiatric Institute, New York, NY, United States, 5 CUNY School of Public Health at Hunter College, New York, NY, United States, 6 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom Background: HIV Counselling and Testing is an important component of HIV prevention and treatment. Early identification of HIV-seropositive status will enable individuals to be linked to HIV care and support. Pathways to Engagement in HIV Care for Newly-Diagnosed South Africans was a prospective cohort study that enrolled newly-diagnosed HIV positive individuals and followed them up for 8 months post diagnosis to explore the barriers and facilitators of enrolling and staying in care. To describe the socio-demographic profile of men and women presenting at PHC clinics for HCT. Methods: Between November 200 and May 202, we screened 2935 individuals from 3 public-sector primary health care clinics in the Durban, South Africa. Women and men who presented for HCT (as opposed to antenatal care) were sequentially approached prior to testing and invited to participate in a screening interview. Eligibility criteria were: 8 years or older, not cognitively impaired, not pregnant, not having previously been diagnosed HIV+, willing to provide written informed consent, to have a witness if illiterate, and to share test results with study staff. Results: Of screened participants, 2746 met the eligibility criteria. Level of education varied, with 38.4% participants having completed Grade 2. Over 9% were unmarried, of which 79% reported having a steady partner. Half of the participants (53.4%) indicated that they were unemployed but were looking for work. Only 6.5% reported being employed full time. Self-motivation for HIV testing was reported by 70.4% of participants, with no difference by gender; 7.4% were motivated by family/friend. However, few reported testing because they were encouraged by the clinic staff. Prior testing was reported by 46%, including 57% of women and 33% of men. Conclusions: Public health benefit would be for HCT programs to tailor interventions for effective gender-based counselling. HIV prevention education should take into account motivations for HIV testing. 75

176 Posters Posters 09: HIV Testing and Counseling P09. P09.2 The Worried Well Among Clients Attending HIV/AIDS Counselling and Testing Services at a Clinical Research Centre in SW Uganda Richard Rwanyonga, Andrew Abaasa, Gershim Asiki, Benjamin Twefeho, Ubaldo Bahemuka, Emanuel Aling, Eugene Ruzagira, Matthew A. Price 2, Anatoli Kamali MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 2 International AIDS Vaccine Initiative, New York, NY, United States Background: HIV/AIDS counselling and testing (HCT) provides a gateway to participation in epidemiological studies. We characterised participants accessing HCT through the two models (facility based [FHCT] vs. community based [CHCT]) offered at the MRC/UVRI clinical research centre in SW Uganda. Methods: From May 200-March 204, we offered HCT to communities (farmers, fisher folk and small townships) in SW Uganda, as part of screening for epidemiological studies. FHCT is offered at the MRC/UVRI clinical research centre in Masaka town where clients voluntarily come for testing, while CHCT participants are approached in their homes or work place and invited to a central point (community hub) within their locality. Both approaches applied the Uganda national HCT guidelines. Results: A total of 23,536 (53% women) individuals (mean age 29 years SD 9.3) received HCT; 9,405 (55% women) mean age 28 years (SD 8.5) FHCT and 4,3 (52% women) mean age 30 years (SD 9.7)CHCT.The main reasons for seeking FHCT were; perceived risk exposure (87%), intending to marry (8%) and poor health (4%). CHCT participants were from fishing communities (78%) and small townships (22%). Over all, HIV prevalence was 3%; [CHCT (6%) Vs. FHCT (9%), p< 0.0], higher among women compared to men (4% vs. 2%; p< 0.0), and higher among those aged >25 years compared to younger age groups (5% vs. 0%, p< 0.0; CHCT 7% vs. 3 %, p< 0.0; FHCT % vs. 7%, p< 0.0). Fishing communities had higher HIV prevalence compared to small townships (8% vs. 8%, p< 0.0). HIV prevalence also varied by the reasons for testing; risk exposure (8.4%), intending to marry (4.6%) and poor health (28.8%). Conclusions: Although majority of those in the FHCT tested because they were worried of their risk, only 8.4% were infected. There is higher burden of HIV in CHCT especially in fishing communities and therefore may be more suitable for recruiting into clinical trials. Does HIV Counseling and Testing Change Sexual Risk Behaviors: Findings from a Community Health Center (CHC) in North Central Nigeria? Ibrahim Suleiman,2, Obianwu Otibho, Jean Njab, Ayodeji Oginni, George Eluwa, Sylvia Adebajo, Babatunde Ahonsi 2, Terfa Kene 3, Jide Keshinro 4 Population Council, HIV and AIDS, Abuja, Nigeria, 2 Population Council, Abuja, Nigeria, 3 Millitary HIV Research Program, Department of Defense, Abuja, Nigeria, 4 Millitary HIV Research Program, Department of Defence, Abuja, Nigeria Background: HIV counseling & testing (HCT) is essential for prevention, care & support. It serves as a means of providing HIV prevention information to people who opt for HCT. The aim of this study was to determine if HCT uptake influences comprehensive HIV knowledge (CHK) & reduces risky sexual behaviors among attendees of a key population focused CHC in Kaduna Metropolis, Nigeria. Methods: From May January 204, clients who accessed sexual health services at a CHC in Kaduna were offered HCT. Data on sexual risk behaviours were collected with the use of a structured HCT client intake form. HIV sero-status was determined using rapid test according to the national algorithm. Descriptive & inferential analyses were conducted to determine the effect of HCT on risk behaviors. Results: A total of 2092 clients accessed the clinic. Majority of the clients were male (98.6%); about 70% were MSM. The median age of the clients was 25 years; above 80% were single; 5% were employed. About 69% of the clients had CHK; 60% had engaged in transactional sex (TS) & 49% had ever been tested for HIV. Comparative analysis between clients with prior HCT uptake & those with no prior HCT uptake, shows that CHK was higher (86% vs. 53%; p< 0.00) among those with prior HCT uptake than those with no prior HCT uptake. TS was lower (43% vs. 75% p< 0.00) while multiple sexual partnerships was higher (87% vs. 57% p< 0.00) among those with prior HCT uptake compared to those with no prior uptake. When controlled for age, sex, occupation, education & marital status, clients with prior HCT uptake were more likely [Adjusted OR= 4.54 (95%CI=3.64, 5.67)] to have CHK & less likely [Adjusted OR= 0.29 (95%CI= )] to engage in TS. Conclusions: HCT uptake among MSM was low; however access to HCT has the potential of increasing CHK as well as reducing risky sexual behaviours among MSM. High quality pre and post-test counselling should be delivered as a means of providing vital behavior change communication messages to MSM. 76 HIV Research for Prevention 204 HIV R4P

177 Posters 09: HIV Testing and Counseling P09.3 Wednesday, 29 October P09.4 LB Young Thai Men who Have Sex with Men and a Voluntary HIV Testing Service, Bangkok, Flash Test Week in Lyon and Rhône Valley: An Expanding Access to Rapid Testing, First Step of the Cascade and Prevention Wipas Wimonsate, Supaporn Chaikummao, Anuwat Sriporn, Pikunchai Luechai, Kesinee Satumay, Santi Winaitham, Somsak Yafant, Sarika Pattanasin, Anupong Chitwarakorn 2, Timothy H. Holtz,3 Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Ministry of Public Health, Department of Diseases Control, Nonthaburi, Thailand, 3 Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and Prevention, Atlanta, GA, United States Background: Previous studies have shown high HIV incidence among young men who have sex with men (YMSM: age 4-2 years) in Bangkok. We investigated the proportion of MSM clients who were YMSM who utilized the anonymous HIV voluntary counseling and testing (VCT) services at the Silom Community Clinic (SCC) during , and the rate of subsequent testing. Methods: We collected data during counselling sessions held between September 30, 2005 and November 30, 203. We used age at first HIV testing as baseline, and the chi-square test for trend for this analysis. HIV testing was voluntary but recommended every 6 to2 months. Results: Between , 6,0 MSM presented for HIV testing. The median age at first HIV testing was 27 years (range: 4-80 years); 2.5%- 7.0% were YMSM during any one year. The proportion of clients who were YMSM who utilized the service during remained stable (p-value for trend 0.47). Among 4,352 who tested HIV-negative at first testing, only 264/692 (38.2%) YMSM had subsequent HIV testing while,452/3,660 (39.7%) older MSM had subsequent testing (p-value 0.65). Conclusions: For the past nine years, the number, but not the proportion, of YMSM who have utilized the free HIV testing service at SCC has increased. The proportion of MSM who returned for subsequent testing did not differ by age group. Specific community efforts must be made to increase the use of VCT services among YMSM. Jean-Michel Livrozet,2, Anne-Cécile Delinotte 2, Sébastien Cambau 2,3, Mireille Joliot-Vilain 2, Colette Coudeyras 2,4, Geneviève Retornaz 2,5, Albertine Pabingui 2,6, Christine Fernandez 2,7, Patrick Caillon 2,8, Aurélie Neveu 9, Véronique Ronzière 0, Yasmine Erraïs 2,, Christine Haydont 2,2, Christian Chidiac 2,3, Pascale Prin 2,4, Jean Paul Godeau 5, Julie Biron 6, Patrice Redon 7, Gérard Millier 2,8, Stéphane Castello 9, Sylvaine Boige-Faure 20, Christelle Bibollet 2, Pascal Pourtau 2,22, Christophe Julien 2,23 Hôpital Edouard Herriot, SMIT-Unité du Pavillon P, Lyon, France, 2 COREVIH Lyon Vallée du Rhône, Lyon, France, 3 ENIPSE, Paris, France, 4 Réseau Virages Santé, Lyon, France, 5 Association de Lutte Contre le Sida, Lyon, France, 6 Datiseni, Lyon, France, 7 Hôpital de la Croix- Rousse, CDAG, Lyon, France, 8 Hôpital Edouard Herriot, CDAG, Lyon, France, 9 Médecins du Monde, Mission France, Lyon, France, 0 Conseil Général du Rhône, Lyon, France, AIDES, Lyon, France, 2 Le Mas, Pause Diabolo, Lyon, France, 3 Hôpital de la Croix-Rousse, SMIT, Lyon, France, 4 Centre Hospitalier de Bourg-en-Bresse, Centre de Santé Publique, Bourg-en-Bresse, France, 5 AIDES, Bourg-en-Bresse, France, 6 Croix- Rouge Française, Bourg-en-Bresse, France, 7 Centre Saliba, Bourg-en- Bresse, France, 8 Hôpital de Privas, CDAG, Privas, France, 9 Agayri, Bourg-les-Valence, France, 20 Mairie de Valence, Direction Santé Famille Environnement, Valence, France, 2 Conseil Général de la Drôme, CDAG le Forum, Valence, France, 22 CRIPS, Lyon, France, 23 Agence Régionale de Santé, Prévention et Promotion de la Santé, Lyon, France Background: The French Minister of Health organized an experimental week of rapid testing in the 4 French regions with high HIV prevalence. In Rhône-Alpes, the action took place in September 203. Methods: Expanding access to rapid testing for the most exposed populations to HIV, Men who have Sex with Men ( MSM), migrants and drugs users ( DU) with: innovating actions in new places next to the targeted populations; new partnerships between hospitals, Voluntary Testing Centers (VTC), community based offers and associations; communication performed by the National Institute for Prevention and Health Education: 400 posters, 000 flyers in gay venues and African shops and commercials in gay and African press and on radios; local coordination by the Regional Committee for HIV ( COREVIH); A questionnaire investigating HIV testing history and sexual behaviors including inconsistent condom use. This questionnaire was analyzed by the French Institute for Public Health Surveillance in June 204. Results: 49 structures were involved: hospitals, VTC, associations and 7 partnerships were signed.38 new people were trained for rapid testing. 049 rapid tests were performed: 60.3% were male, 39.5% female and 0.2% trans. 60.5% were between 8 and 30 year-old. 74.5% were born in France and 20.3% in a foreign country:.8% in sub-saharan Africa,4.5% in North Africa.9.6% were MSM, 3.5% DU. 55.8% reported non-condom use, 6.5% sex for money, 4% drug use in the past 5 years. 64.3% have had an HIV test before and 35.5% in the past 2 years. 27.2% of people performed a HIV test for the first time. In total, 4 tests were positive (0.4%):3 migrants and MSM, and the patients were referred to HIV units due to the links developed before this week. Conclusions: This week is an original initiative to enhance HIV prevention and screening in high-risk populations. An important proportion of persons were HIV tested for the first time. Finally, it allows new partnerships between community-based associations and referral centers. 77

178 Posters Posters 09: HIV Testing and Counseling P09.5 LB Assessment of Namibian Pharmacy Practices of the Sale of Over-the-Counter HIV Rapid Test Kits in the Private-sector Matthew Rosenthal, Ismelda Pieterson 2, Cheryl Johnson 3, Nersesian Paula 4, Mechtild Hulsman 5 USAID Namibia, HIV Office, Windhoek, Namibia, 2 Government of Namibia, Ministry of Health and Social Services, Windhoek, Namibia, 3 World Health Organization, HIV Testing and Counseling, Geneva, Switzerland, 4 AIDSTAR, John Snow International, HIV, Arlington, VA, United States, 5 Consultant, Windhoek, Namibia P09.6 LB ARV-based Prevention for Women: New Data on HIV Testing Behaviors from Potential Users of Vaginal Microbicides in Mpumalanga, South Africa Saiqa Mullick, Martha M. Brady 2, Barbara A. Friedland 2, Thabiso Mango 3, Ravikanthi Rapiti 4 Wits Reproductive Health and HIV Institute (WRHI), Johannesburg, South Africa, 2 Population Council, New York, NY, United States, 3 Solutions for Innovative Policies, Programs, and Technologies (Solutions-IPPT), Johannesburg, South Africa, 4 Population Council, Johannesburg, South Africa Background: In 20, the Namibia private sector began the sale of Overthe-Counter (OTC) HIV rapid test kits (RTK) for personal-use. By beginning 202, volume of RTK sales increased within the general public. In Dec 202, a team conducted a rapid survey of RTK for personal-use. The GRN has identified increased HIV Testing and Counseling (HTC) as an entry point into HIV care and treatment. Despite high HIV prevalence (3.4%), only 54% of Namibia s population has received HTC. While, RTK may offer a new strategy to HTC programs for implementing mixed HTC models, concern within GRN on unregulated sale and potential social harms of RTK remain. Methods: The assessment was conducted over a month. Key informants (n=68) included private-sector and GRN staff. A survey at 53/2 pharmacies (48%) throughout 6 regions in Namibia was administered to pharmacists. All of the pharmacies in 5 high prevalence regions were surveyed and a sample in Windhoek. Survey assessed attitudes, practices and approaches. 52/ 53 pharmacies participated and 36/52 reported current RTK sales. Results: Analysis of RTK varied by pharmacy and geography. Some coastal city pharmacies reported 30 tests sold in the last month prior to the survey and southern areas reported lower sales (-3 in same period). Price and location of tests differed (i.e. behind counter, controlled sales, on pharmacy floor) and influenced sales volume. Attitudes, practices and training of pharmacists varied and reflected poor regulation. Pharmacists (38%) reported having been trained in performing HIV RTK and 62% reported HTC training, ranging from pre-service to self-training. Stocking and supplying RTK varied with good lead time (i.e. day in some locations), but little quality assurance, maintenance and proper storage. 4 different test kits were sold to the public. Conclusions: Additional assessments of RTK in Namibia need to be conducted; including development of regulatory frameworks for RTK in Namibia and user acceptability, feasibility and quality of RTK. Background: Use of ARVs, such as Tenofovir (TFV) gel, for preexposure prophylaxis (PrEP) will require regular HIV testing to ensure users are uninfected before initiating and continuing product use. Understanding HIV testing practices among potential users is critical in planning for introduction. Methods: A cross-sectional descriptive study of women s healthseeking behaviors and HIV-testing practices was conducted among a convenience sample of women attending 5 high-volume primary health clinics (PHC) in Mpumalanga, South Africa in 204. A quantitative faceto-face interview captured information on frequency and circumstances around potential HIV exposure, HIV testing, risk perception, awareness of and willingness to try TFV gel. Results: A total of,227 women completed the survey. 93% of women reported having been tested at least once and 90% who tested more than once did so in the past year. The vast majority (over 90%) knew their HIV status, and the majority of those (79%) disclosed this information to someone. Although most women were not familiar with microbicides, after being provided with information 94% said they would be willing to try TFV gel. Most women (92%) said they would prefer to get the product from a clinic and 8% preferred receiving it from a nurse. The majority (82%) believed that a vaginal gel used only around the time of sex would be easier to use than a daily oral pill. Half said they would be willing to get tested for HIV every 3 months and an additional 22% said they would be willing to get tested once a month. However, 68% indicated that they would only use such a product if it were free. Conclusions: The data show that many women are already being tested regularly for HIV, a pre-requisite for provision of ARV-based prevention. Women s stated willingness to try TFV gel suggests an overall positive attitude. Preferences for accessing microbicides from a nurse at a clinic suggest that delivery of TFV gel from existing primary health services is acceptable. 78 HIV Research for Prevention 204 HIV R4P

179 Posters 09: HIV Testing and Counseling P09.7 LB Wednesday, 29 October P09.8 LB The Cost of Community Based HIV Counseling and Testing and Linkage to Care in Rural South Africa: Estimates from the Linkages Randomized Control Trial Monisha Sharma, Heidi Van Rooyen 2, Connie Celum 3, Jared Baeten 3, Carol Levin 3, Ruanne Barnabas 3 University of Washington, Epidemiology, Seattle, WA, United States, 2 Human Sciences Research Council, HIV/AIDS, STIs and TB, Pietermaritzburg, South Africa, 3 University of Washington, Global Health, Seattle, WA, United States Background: Community based HIV testing and counseling (HTC) and linkage to HIV care has demonstrated high effectiveness. However, HTC program costs are needed to inform policy decisions. Methods: We estimated costs of mobile clinic and home-based HTC with point-of-care (POC) CD4 testing and counselor follow up at home to encourage linkage. Costs were collected in KwaZulu-Natal in 203 from the Linkages Study, a randomized trial of community HTC and linkage to care. Time and motion studies were conducted to separate research from program activities. Costs were obtained from budgets, invoices and staff interviews. We assumed task shifting from nurses to community workers. Program effectiveness was estimated from our pilot study in a nearby area (N=272, 30% HIV prev). Total program costs for HIV+ persons were divided by number virally suppressed at 2 mos to estimate incremental cost per person virally suppressed. Results: Program cost of mobile HTC was $5.45 per HIV- person tested and $8.28 per HIV+ tested. POC CD4 increased cost per HIV+ tested to $4.78 and POC with follow-up cost $2.78. Home HTC cost $8.22 and $2.3 per HIV- and HIV+ person tested. Using effectiveness from our prior study (32% of HIV+ persons were ART eligible at CD4 350mL, of whom 69% initiated ART and 70% virally suppressed at 2 mos), incremental cost for all HIV+ persons for home HTC with POC CD4 and counselor follow up was $26.0 per person linked to care and $79.9 per person virally suppressed. Mobile HTC with POC CD4 and follow-up cost $96.62 and $37.30 per person linked to care and virally suppressed. Assuming similar program effectiveness under the new ART guidelines of eligibility at CD4 500mL, costs would decrease to $00.95 and $77.35 per person virally suppressed through home and mobile HTC respectively. Conclusions: Community HTC with POC CD4 and follow-up achieves high linkage and viral suppression at costs of $37-79 per HIV+ person virally suppressed. Incremental costs are expected to decrease with new ART guidelines. Does Culture Affect Counselling in HIV Prevention Research? Colleen Herman, Annalene Nel International Partnership for Microbicides, Clinical Operations, Cape Town, South Africa Background: The Ring Study is a Phase III microbicide study, currently enrolling in communities with a culturally-diverse background in Southern and Eastern Africa. The person-centred counselling (PCC) approach, which stimulates probing, is used by Research Centre (RC) counsellors. Participants and counsellors alike are diverse in ethnicity, cultural background and education. Culture can be described as the blue print for who we are. It includes patterns, beliefs, values, expectations and symbols for how people behave, think and feel in a certain social group. PCC aims at improving retention to study visits and adherence to study product. Methods: The following counselling is conducted during The Ring Study: ring use adherence; HIV pre- and post-test counselling; risk reduction; contraception and visit adherence counselling. RC staff attended informal training sessions on understanding and implementing the PCC approach. Regular assessments are conducted to get feedback on the implementation of the approach; e.g. visiting RCs; engaging with counsellors and nurses who conduct counselling on site. Additional feedback on adherence is obtained subjectively via questionnaire feedback. Results:. Participants differ and decisions are influenced by cultural norms (e.g. partner, family, friends and community). 2. Cultural self-awareness is an important counselling aspect as it allows identification of differences and more empathy and sensitivity in the counselling relationship. 3. Reticence in probing for in-depth PCC answers. Conclusions: Cultural norms are powerful and can sway a participant from her good intentions, even if the principles of the PCC approach are used effectively. These cultural norms can result in different outcomes with regard to protocol compliance and ring adherence. Further qualitative research is recommended to determine if culture affects counselling in HIV prevention research as this could lead to improved retention and adherence. 79

180 Posters Posters 0: Immunogens P0.0 Improving the Vaccine that Provided Protection in the RV44 Trial P0.02 Canyon Shielding of the CD4-Binding Site on HIV- Trimer Javier F. Morales, Trevor J. Morin, Bin Yu, Gwen P. Tatsuno, David L. Alexander, Sara M. O Rourke, Richard Theolis, Kathryn A. Mesa, Phillip W. Berman,2 University of California, Santa Cruz, Biomolecular Engineering, Santa Cruz, CA, United States, 2 Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA, United States Background: Two approaches have highlighted the importance of antibodies to the gp20 V/V2 domain in providing protection from HIV- infection. First, the RV44 HIV- trial documented a correlation between non-neutralizing antibodies to the V2 domain and protection. Secondly, multiple broadly neutralizing monoclonal antibodies (bnmabs) to glycan-dependent epitopes (GDEs) in the V/V2 domain have been isolated from rare infected individuals (elite neutralizers). Methods: Analysis of the glycan content of the A244 and MN-rgp20 antigens used in the RV44 trial showed that they lacked the glycans required for the binding of bn-mabs to the V/V2 and the V3 domains. We wondered if we could improve the ability of these immunogens to elicit both types of protective antibodies by in vitro mutagenesis and by production in GnTI- cells that limit carbohydrates to mannose-5 glycans. Results: We found that we could re-engineer monomeric gp20s used in the AIDSVAX B/E vaccine used in the RV44 trial to bind multiple bnmabs, e.g. PG9, PGT28, and VRC0 with high affinity. We also found that we could produce stable disulfide bonded fragments of the V/V2 domain (scaffolds) that preserved the 4-stranded beta sheet structure and could bind PG9 and PG9-like antibodies. Immunization studies showed that these gp20s and scaffolds could significantly enhance the magnitude of antibody responses that correlated with protection in the RV44 trial. These immunogens also enhanced the magnitude of antibodies directed to the PG9 binding site. Conclusions: Our studies suggest that we can substantially improve the immunogenicity of the vaccine that provided protection in the RV44 trial. Our goal is to improve the level of protection (3.2%) observed in RV44 to a level of 60% or more required for regulatory approval. By improving an existing vaccine with a record of safety in more than 9000 subjects and demonstrated efficacy, years of time and millions of dollars can be saved compared to developing a new vaccine from scratch. Lei Chen, Peter D. Kwong VRC/NIAID/NIH, Bethesda, MD, United States Background: Many viruses use single-headed immunoglobulin domains as receptors, but restrict access of double-headed immunoglobulins that comprise the recognition domains of human antibodies. The membranedistal single-headed immunoglobulin domain of CD4, for example, is recognized by the HIV- gp20 envelope glycoprotein, but access to the site of CD4 binding is restricted for most human antibodies. Methods: To understand the immunological consequences of doubleheaded versus single-headed recognition, we determined crystal structures of gp20 in complex with four single-headed antibodies, A2, C8, D7 and J3, derived from immunized llamas, which target the CD4- binding site. Results: Antibody J3 neutralization breadth approached 98% despite a targeting precision similar to that of the double-headed antibody VRC6, which has only ~56% breadth. A2 breadth was 4% despite a targeting precision below that of b3 and F05, which have only ~0% breadth. Double-headed A2 lose neutralize activity against twenty selected viruses. Conclusions: Overall, HIV- neutralization by double-headed antibodies at the CD4-binding site required substantially higher targeting precision, and more V-gene affinity maturation than single-headed antibodies, an effect our results attribute to canyon shielding of this important vaccine target. 80 HIV Research for Prevention 204 HIV R4P

181 Posters 0: Immunogens P0.03 Wednesday, 29 October P0.04 Optimization of a Clade A Env Outer Domain for Enhanced Binding to Germlines of Diverse VRC0 Class Antibodies Cheng Cheng, Ivelin Georgiev, M Gordon Joyce, Xuejun Chen, Adam Bossert, Wing-Pui Kong, Peter D. Kwong, John R. Mascola Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States Negative Selection Using CD4-binding Site Non-broadly Neutralizing Antibodies Yields Conformationally Homogeneous Clade B and C SOSIP Trimers Javier Guenaga, Natalia de Val 2, Karen Tran, Karen Satchwell, Yu Feng, Andrew Ward 2, Richard Wyatt International AIDS Vaccine Initiative (IAVI), Neutralizing Antibody Center, Scripps Research Institute, La Jolla, CA, United States, 2 Scripps Research Institute, Immunology and Microbial Science, La Jolla, CA, United States Background: Effective HIV vaccines need to elicit broadly neutralizing antibodies (bnab) as an essential component of protective immunity. Among the known bnab, those that target the conserved CD4 binding site (CD4bs) are among the most broad and potent. The VRC0 class of CD4bs antibodies derive from a VH-2 germline gene and have a characteristic short (5AA) CDRL3. These antibodies are highly affinity matured and when reverted to germline sequence fail to recognize most HIV Env sequences. Thus, a major challenge is to design immunogens that would activate the appropriate naïve B cells and generate VRC0 class antibodies. Previous studies have identified few clade C gp40 and clade B gp20 outer domain constructs (OD) that can engage germline of a subset of CD4bs antibodies. Here, we aim to identify HIV clade A OD mutants that can bind to germlines of diverse CD4bs antibodies to be used as HIV imunogens. Methods: Clade A OD4.2.2 (PDB code 4I3R) was displayed on the cell surface using a yeast display system. Mutations in its D loop ( ), which constitutes part of the CD4bs, were introduced by PCR to construct two libraries: one with randomized sequences and the other with sequences designed by structure based bioinformatics. The libraries were screened for binding to germline revertants of VRC0-class antibodies. Germline binders were sorted with FACS and characterized. Results: Consistent with previous studies, the original yeast displayed OD failed to bind to any of the germline revertants of CD4bs antibodies. Two clones from the randomized D loop library were identified to bind VRC03 germline (VH-2). When displayed on lumazine synthase 60mer nanoparticles, they bound to several VRC0 class germline revertants including VRC0, VRC07, VRC20, 2A2, CH3 and VRC03. Conclusions: Optimization of amino acids in loop D of the OD helped to identify mutants that can bind to diverse VRC0 class germline reverted antibodies. These OD constructs can thus serve as priming immunogens to elicit CD4bs bnab. Background: The recently published structures of the clade A-derived BG505 SOSIP trimers, mimetics of the native HIV envelope glycoprotein (Env) spike, mark the beginning of new era in HIV structural biology. Displaying a well-ordered quaternary array, the BG505 SOSIP trimers display an excellent antigenic profile, discriminating recognition by broadly neutralizing antibodies (bnabs) from non-broadly neutralizing antibodies (non-bnabs) and represent interesting Env-based immunogens. Even with this significant advance, obtaining soluble SOSIP trimers derived from other clades has been challenging. Here, we report the isolation of two SOSIP trimers derived from clades B and C. Methods: We modified the 2G2 affinity purification by replacement with lectin affinity purification, affording a scalable process with mild elution conditions. Using CD4 binding site-directed (CD4bs) non-bnabs in a negative selection purification process, we obtained homogeneous wellordered clade B JRFL and clade C 6055 SOSIP trimers from a mixture of conformations. We then used EM, bio-layer interferometry and differential scanning calorimetry to characterize these new Env mimetics. Results: Following negative selection, by EM we demonstrated that we achieved nearly complete removal of disordered trimers, recovering predominantly well-ordered trimers. We obtained EM 3D reconstructions of the trimers, unliganded, in complex with scd4 and the bnabs, VRC0, VRC03 and the newly identified trimer-specific PGT5. We employed bio-layer light interferometry to get a full antigenic profile and showed that the negatively selected homogeneous trimers behave as faithful mimetics of the native spike, possessing avid recognition by bnabs and poor recognition by non-bnabs. Conclusions: This study affords a new means to obtain conformationally homogeneous soluble mimetics of Env derived from two different HIV clades from a mixture of Env conformations that can potentially expand to other strains. 8

182 Posters Posters 0: Immunogens P0.05 Double Safety Measure for Vaccines Based on Replication Competent Poxvirus P0.06 CD8 T-cell Based HIV Vaccines - Is Targeting Conserved Epitopes the Answer? Ying Liu, Qicheng Zhang, Shuhui Wang, Chang Liu, Zheng Liu, Hong Peng, Yiming Shao National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Division of Virology & Immunology, Beijing, China Background: Poxvirus is a commonly used viral vector in vaccine research. There is no pre-existing vector immunity in young population due to eradication of the smallpox in 970s. Replication competent pox vector is more immunogenic than non-replicating one, but may not be safe in immunecompromised people. In HIV vaccine research, double safety measures may be needed to deal with rare case of spill over from vaccinee to AIDS patients. The Vaccinia Virus Tiantan (TV), the Chinese small pox vaccine, was attenuated and used as vector for recombinant HIV- vaccine (rtv). The rtv has proved to be safe and immunogenic in phase I /II trails in China. Methods: To provide a double safety measure, the Herpes Simplex Virus thymidine kinase gene was inserted into the TV to obtain the replication competent TV/tk vector. The inhibition of the Ganciclovir (GCV) to the viral vector was tested in vitro and in vivo. The immunogenicity of TV/tk vector was also tested and compared with the rtv. Results: The replication of TV/tk on CEF and Vero cells were significantly inhibited with EC 50 of 5.4 µm and 2.5 µm respectively.gcv could inhibit the replication of TV/tk in brains of mice and provide 00% protection to the mice at a dose 80mg/kg/day. If the GCV treatment began before 3 day p.i., mice infected with TV/tk could be protected. The TV/tk/Env can stimulate strong humoral and cellular responses, similar to that by TV/Env vector. Conclusions: The study showed that insertion of HSV-tk gene into TV vector is a reliable rescue measure to the replication-competent vaccinia virus vaccination. This strategy can provide a double safety guarantee in large scale application of such vaccine in situation, where the vaccine target population is mixed with ineligible immune compromised individuals. Shelby L. O Connor, Dane Gellerup, Max Harris, Ericka Becker University of Wisconsin-Madison, Madison, WI, United States Background: Immunodominant CD8 T cell responses emerge in the first few weeks after HIV/SIV infection, suppress virus replication, and select for escape variants. Although T cell based HIV vaccines have not successfully provided sterilizing immunity, vaccine-elicited CD8 T cells may contribute to the control of replication of breakthrough viruses. It is critical that a T cell based vaccine generates an effective pool of memory CD8 T cells that are available to respond during acute HIV infection and effectively control both acute and chronic virus replication. Designing a vaccine to elicit these potent CD8 T cells in all vaccinated individuals, however, is a daunting challenge. Conceptually, a vaccine that elicits CD8 T cell responses targeting highly conserved virus peptide sequences would have the most widespread impact, but the efficacy of T cells targeting these conserved epitopes is unknown. Methods: We employ a model of SIVmac239Δnef-infected MHCidentical Mauritian cynomolgus macaques to test the hypothesis that CD8 T cells targeting conserved epitopes are unable to detect and destroy virally infected cells. Accordingly, we expect that CD8 T cells targeting epitopes that accumulate variants are more effective at controlling virus replication. To test this hypothesis, we are creating variants of live attenuated SIVmac239Δnef designed to elicit CD8 T cells targeting epitopes that do and do not accumulate variants. Results: We will determine whether the mutant viruses are fit and whether the included variant epitope sequences are no longer detected by CD8 T cells. In the future, we will determine if acute CD8 T cells targeting these different categories of epitopes are able to control virus replication, in vivo. Conclusions: The conclusions from this study will help identify whether CD8 T cells that develop during acute HIV infection can be specific for highly conserved epitopes and whether they can control virus replication. 82 HIV Research for Prevention 204 HIV R4P

183 Posters 0: Immunogens P0.07 Wednesday, 29 October P0.08 Bivalent NYVAC-based Vaccine Candidates against HIV/AIDS Expressing Clade C Trimeric Soluble gp40(zm96) and Gag(ZM96)-Pol- Nef(CN54) as VLPs Beatriz Perdiguero, Carmen Elena Gómez, María Victoria Cepeda, Lucas Sánchez-Sampedro, Juan García-Arriaza, Ernesto Mejías-Pérez, Victoria Jiménez, Cristina Sánchez, Carlos Oscar S. Sorzano, Julie Delaloye 2, Thierry Roger 2, Thierry Calandra 2, Ralf Wagner 3, Benedikt Asbach 3, Karen V. Kibler 4, Bertram L Jacobs 4, Giuseppe Pantaleo 2, Mariano Esteban Centro Nacional de Biotecnología, CNB-CSIC, Madrid, Spain, 2 Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland, 3 University of Regensburg, Regensburg, Germany, 4 The Biodesign Institute at Arizona State University, Tempe, AZ, United States Background: The generation of vaccine candidates against HIV/AIDS able to induce long-lasting protective immunity remains a major goal in the HIV field. The reduced efficacy (3.2%) against HIV infection observed in the RV44 Thai clinical trial highlighted the need to develop novel improved poxvirus-based recombinants. Methods: In the present study, we have generated two novel NYVAC vectors expressing HIV- clade C gp40(zm96) (NYVAC-gp40) or Gag(ZM96)-Pol-Nef(CN54) (NYVAC-Gag-Pol-Nef) and defined their biological characteristics in cultured cells and in mice. Results: Insertion of the HIV- genes in the viral genome does not affect the replication capacity of the NYVAC recombinants in primary chick cells and the HIV- antigens are correctly expressed and released from the cells, with Env protein as a trimer (NYVAC-gp40), while in cells infected with NYVAC-Gag-Pol-Nef, Gag-induced VLPs are abundant. Electron microscopy revealed that VLPs are accumulated with time at the cell surface, with no interference with NYVAC morphogenesis. Gag- Pol-Nef expression in human primary monocytes markedly increases the levels of immunomodulatory molecules, such as cytokines and chemokines. Both recombinant viruses show an attenuation profile in immunocompromised BALB/c mice after intracranial inoculation. Analysis of the immune responses elicited in mice after homologous NYVAC prime/nyvac boost immunization shows that the two recombinant viruses induced polyfunctional Env-specific CD4 or Gagspecific CD8 T cell immune responses. Antibody responses against gp40 and p7/p24 were also elicited by both NYVAC vectors. Conclusions: Our findings showed that bivalent NYVAC vectors can be considered as candidate vaccines against HIV/AIDS. Characterization of the Binding Affinity of Siglec- to gp20, gp45, and V2 Loop via Sialic Acid Binding Motif Hung V. Trinh, Ousman Jobe, Guofen Gao 2, Carl R. Alving 3, Venigalla Rao 2, Mangala Rao 3 U.S. Military HIV Research Program (MHRP)/HJF, Laboratory of Adjuvant and Antigen Research, Silver Spring, MD, United States, 2 Catholic University of America, Department of Biology, Washington, DC, United States, 3 U.S. Military HIV Research Program (MHRP), Walter Reed Army Institute of Research, Laboratory of Adjuvant and Antigen Research, Silver Spring, MD, United States Background: Although HIV- primarily infects T-cells through the interaction of viral envelope with CD4 and a co-receptor CCR5/CXCR4, CD4 expresses in low level in macrophages. Recently, sialic acid-binding Ig-like lectin (Siglec-, CD69), which is highly expressed on the cell surface of macrophages, has been identified as a major receptor for HIV- infection. It is known that Siglec- binds to glycans containing sialic acid. To further understand the mechanism; we determined the binding affinity/kinetics of monomeric gp20 and trimeric gp45 to assess if there were differences in their ability to bind to Siglec-. We further narrowed down the env-binding motif to the V2 loop of gp20. Finally, we conducted an inhibition assay in the presence/absence of lactose (LA), 2, 6 -Sialyllactose (6 -SL) and sialic acid (SA) to identify the nature of the interactions. Methods: Siglec- was immobilized on a CM5 chip on a Biacore T200. JRFL and SF62 (clade B) gp20, gp45, and V2 loop proteins were captured. In additional experiments, these env proteins were immobilized, while Siglec- was injected over that surface. For the inhibition assays, LA, SL and SA were mixed with the env proteins or with Siglec- and then injected onto the immobilized Siglec- or env proteins, respectively. Results: The gp20 and gp45 binds to Siglec- with high affinity, 4.3 nm (JRFL gp20), nm (JRFL gp45) and nm (SF62 gp45). Importantly, we found that the interaction of env protein to Siglec- took place through the V2 loop, (4.34 nm JRFL V2;.96 nm SF62 V2). The presence of SA completely blocked the interaction of the env and the V2 loop protein with Siglec-, suggesting thereby that the interaction occurred via a SA motif. Conclusions: The binding of gp20, gp45, and V2 loop to Siglec- is a high affinity interaction. Binding occurs through the V2 loop and requires sialic acid. This suggests that the vaccine design should aim to induce antibodies against V2-sialic acid complexes to prevent HIV- infections. 83

184 Posters Posters 0: Immunogens P0.09 P0.0 Scalable and Robust Purification of Intact HIV- gp20 Monomer Subunit Antigens Yingxia Wen, Sai Tian, Christine Linton, Susan Barnett, Andrea Carfi Novartis Vaccines, Cambridge, MA, United States Background: Development of an effective vaccine against HIV- is challenging due to various viral evolutionary mechanisms to evade human immune system. The partial efficacy of the recent RV44 vaccine efficacy trial in Thailand provides hope for improvements of vaccine regimens for higher efficacy. Clinical trials in Thailand is planned to confirm and extend the results of the RV44 trial with the vaccine strategy of poxvirus vector prime plus envelope protein boost. Methods: To produce gp20 monomers, we generated CHO stable cell lines, consistently expressing intact gp20 subunits with high yield. Simple, scalable and robust antigen purification processes were developed to generate gp20 proteins with high purity, integrity and homogeneity. Results: The ion-exchange based purification strategy enabled the isolation of gp20 from host cell contaminates and separation of gp20 monomer from dimer. This purification strategy also significantly removed contaminated proteinase and inhibited proteinase activity, leading to intact gp20 monomer with high purity. Purified gp20 monomers were stable, either alone or in combination, and when formulated with adjuvant Alum and MF59. The antigenicity and immunogenicity studies are on-going. Conclusions: Scalable and robust purification process of intact HIV- gp20 monomer subunit antigens is developed to produce HIV gp20 antigen for clinic trials. Transmitter Founder Multi-envelope DNA Vaccine Induces Potent Cross-clade Cellular and Humoral Responses in Rabbits and Nonhuman Primates Megan Wise, Muthumani Karuppiah, Janess Mendoza 2, Natalie Hutnick, Jian Yan 2, David Montefiori 3, Celia Labranche 3, Kate Broderick 2, Matthew Marrow 2, Niranjan Sardesai 2, David Weiner University of Pennsylvania, Philadelphia, PA, United States, 2 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States, 3 Duke University, Durham, NC, United States Background: It has been reported that guinea pigs vaccinated with transmitted founder gp40 envelope proteins are able to induce low neutralizing antibodies with some improved breadth (Liao et al 203). This general induction of coverage may be ideal for a priming immunization, establishing a response which is able to be boosted with the addition of either chronic or consensus envelopes. Since the DNA vaccine platform is seen as a priming regiment, we aimed to investigate if broad responses could be induced using primary transmitter founder (TF) and acutely isolated gp60 immunogens developed from clades A, B, and C as DNA plasmids. Methods: Initial studies were performed in rabbits, which were immunized by EP route with different combinations of synthetically optimized gp60 immunogens from clades A, B, and C. Animals received the same amount of total DNA and were immunized at 3 week intervals. Humoral responses were determined after each immunization. As a follow on, non-human primates were immunized with clusters of gp60 DNA at weeks 0, 4, 8, 2 and boosted at week 48. Results: Rabbits immunized with clusters of clade A gp60 envelope DNA were able to induce cross-clade binding titers with limited neutralization. Including TF envelopes from different clades increased binding titers and neutralization breadth and potency. NHP immunized with clusters of clade A and B gp60s showed cross-clade cellular responses after two immunizations. These responses increased after each immunization and were maintaining into memory. After two immunizations, NHP were able to induce cross clade Ab binding titers against primary gp20 from clades A, B and C and tier neutralization. Conclusions: DNA plasmids encoding TF and acute gp60 immunogens are expressed and induce a potent immune response in vivo. We observed for the first time that exposure of the immune system to multiple DNA env vaccines at one time dramatically alters the immune phenotype induced resulting in increased magnitude and breath of responses. 84 HIV Research for Prevention 204 HIV R4P

185 Posters 0: Immunogens P0. CD4-binding-Site Recognition by VH-46 Germline-derived HIV- Neutralizers Wednesday, 29 October P0.2 Longitudinal Antibody Development in SHIV AD8 Infected Non-Human Primate Priyamvada Acharya, Tongqing Zhou, Cinque Soto, Lei Chen, Timothy S. Luongo, Stephanie Moquin, Ivelin S. Georgiev, Stephen D. Schmidt, Mark K. Louder, M. Gordon Joyce, Yongping Yang, Baoshan Zhang, Johannes Scheid 2, Michel C. Nussenzweig 2, John R. Mascola, Peter D. Kwong NIAID, NIH, VRC, Bethesda, MD, United States, 2 Rockefeller University, New York, NY, United States Background: The human immune system generates diverse antibodies against the binding site for the CD4 receptor in response to infection or vaccination. Two classes of antibodies that effectively neutralize HIV- through CD4 mimicry have been described, including those derived from VH-2 germline, which include the well-characterized VRC0 class of antibodies, and those derived from VH-46. Methods: We defined structural modes of recognition of three VH- 46 germline-derived antibodies from 2 donors by crystallizing antigenbinding fragments in complex with extended core version of the HIV- gp20. We used surface plasmon resonance and biolayer interferometry to determine antigen-binding properties, tested neutralization in a representative panel of 92 viruses, and performed cross-donor phylogenetic analyses to study antibody evolution. Results: VH-46 derived antibodies show a structural mode of CD4 mimicry distinct from that of the VH-2 derived antibodies. The altered mode of heavy chain recognition allows the VH-46 derived antibodies to accommodate light chain CDR3s of different lengths. Cross-donor phylogenetic analysis showed that the VH-46 derived antibodies from 2 donors evolved similarly and indicated that the VH-46 antibodies form a class. Conclusions: Our studies show how antibodies of the VH-46 class achieve CD4 mimicry, expand the range of known structural solutions that permit such heavy-chain mimicry, and reveal how small differences in germline (e.g. between VH-2 and VH-46) can impact mature antibody recognition. Zizhang Sheng, Joseph R. Francica 2, Yoshiaki Nishimura 3, Stephen D. Schmidt 2, Rebecca Lynch 2, Sam Darko 2, Zhenhai Zhang,4, Frederick Jaeger 5, Munir Alam 5, Daniel Douek 2, John R. Mascola 2, Malcolm A. Martin 3, Robert A. Seder 2, Lawrence Shapiro Columbia University, Department of Biochemistry and Molecular Biophysics, New York, NY, United States, 2 National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States, 3 National Institute of Allergy and Infectious Diseases, National Institutes of Health, Laboratory of Molecular Microbiology, Bethesda, MD, United States, 4 Southern Medical University, Division of Nephrology, Nanfang Hospital, Guangzhou, China, 5 Duke University Medical Center, Duke Human Vaccine Institute, Durham, NC, United States Background: SHIV infected Non-human primate (NHP) is an important system to study antibody development relevant to HIV- infection in humans. CCR5-tropic SHIV AD8-EO is a clade B virus that has a tier 2 neutralization phenotype and can induce broadly neutralizing antibodies. However, the developmental characteristics of SHIV AD8 elicited NHP antibodies are unclear. Methods: Four of eight SHIV AD8 infected Rhesus macaques developed cross-reactive antibodies (good neutralizers) and others showed weak Tier virus neutralizing activity (poor neutralizers). Peripheral memory B cells from all animals at four time frames (6-8, 26-32, 52-54, and 9-0 weeks) were sorted into gp20 reactive (gp20+) and nonreactive B cells (gp20-). Antibody heavy chains were then sequenced by 454-pyrosequencing. Germline V gene was assigned for each antibody sequence using a newly characterized NHP heavy chain V gene database. Somatic hypermutation level and CDRH3 length were then calculated using an in-house bioinformatics pipeline. Results: Compared with gp20- B cells, VH gene composition analysis of gp20+ B cells showed enriched VH3-J (human ortholog VH3-23), VH4-D (human VH4-B) and VH4-A (human VH4/OR5-8) antibodies from good neutralizers. The somatic hypermutation level of gp20+ B cell antibodies increased gradually from ~4% at week6 to ~0% at week0. In gp20+ B cells of six animals, more frequent usage of antibodies with CDRH3 ~20aa in length is observed. In animal DCF (good neutralizer), more than ten long CDRH3 (>=28aa) antibody lineages were elicited at early time points and some continued expanding through week08 post infection. Conclusions: SHIV AD8 infection preferably elicited certain VDJrecombined antibodies. The SHIV AD8 infected NHP system captures characteristics of antibody development in HIV- infected humans, such as longitudinal antibody maturation and elicitation of long CDRH3 antibodies. This supports the idea that SHIV AD8 infected NHP provides a good model for the study of HIV antibody development. 85

186 Posters Posters 0: Immunogens P0.3 LB A Novel Trimeric VV2-Scaffold Immunogen Induces V2q-Specific Antibody Responses Xunqing Jiang, Max Totrov 2, Constance Williams 3, Wei Li 4, Shan Lu 4, Shixia Wang 4, Susan Zolla-Pazner 3, Xiang-Peng Kong NYU School of Medicine, Department of Biochemistry and Molecular Pharmacology, New York, NY, United States, 2 Molsoft LLC, San Diego, CA, United States, 3 NYU School of Medicine, Department of Pathology, New York, NY, United States, 4 University of Massachusetts Medical School, Department of Medicine, Worcester, MA, United States P0.4 LB Chimeric Bovine-V-region and Human-Cregion mabs with Long and Extensively Mutated CDHR3 Domains Bind HIV- Env gp40 Trimers, but Not gp20 Monomer Behnaz Heydarchi, Robert Center, Sri Ramarathinam 2, Christopher Gonelli, Brian Muller,3, Charlene Mackenzie, Jack Cuthbertson, Marit Kramski, Damian F.J. Purcell University of Melbourne, Microbiology and Immunology, Melbourne, Australia, 2 Monash University, Biochemistry and Molecular Biology, Clayton, Australia, 3 Reef Pharmaceutical, Melbourne, Australia Background: Data from the RV44 vaccine clinical trial revealed that high levels of antibodies to the first and second variable regions (VV2) of gp20 were correlated with the modest protection from HIV- infection; thus VV2 is a potential target for HIV/AIDS vaccine development. VV2 is known to harbor three distinct epitope types including V2q defined by quaternary broadly neutralizing mabs such as PG9/PG6, V2p defined by the RV44 mabs CH58/CH59, and V2i defined by a panel of mabs, such as 697-D, targeting a region overlapping the VV2 integrin-binding site. It is highly desirable to develop immunogens that can induce antibody responses specific for these epitope types. Methods: We engineered a trimeric VV2 (ZM53 sequence) immunogen that was designed to mimic Env conformation by inserting it into a trimeric scaffold (PDB ID 2J9C). We produced this immunogen in 293 cells and tested its antigenicity by ELISA. Rabbits were immunized by the DNA prime-protein boost regimen with a gp20 DNA (ZM09 sequence) and the VV2 ZM53-2J9C immunogen. The immunogenicity was then assessed by antibody competition assays. Results: We found that the VV2-2J9C immunogen could bind mab PG9, CH58 and 697-D, thus it harbors the V2q, V2p and V2i epitopes. We also found that this immunogen is highly immunogenic in rabbits. Direct binding competition ELISA assays showed strong competition between immune rabbit sera and CH58 or PG9, but not with 697-D, thus this trimeric VV2 immunogen induced strong antibody responses targeted not only the linear V2p epitope type but also the quaternary V2q epitope type. Conclusions: Our results demonstrate that structurally constrained VV2 scaffolds with Env quaternary conformation can be designed and synthesized, resulting in elicitation of antibody responses with specificities overlapping the PG9 epitope region - a first step in immunogen design to target a major vulnerable site of HIV- Env. Background: Bovine immunoglobulins (Ig) typically have variable third heavy complementarity determining regions (CDRH3) for antigen engagement that are significantly longer than in humans and other mammals. We aimed to isolate HIV- bovine memory B cells binding HIV- AD8 Env gp40 trimer antigen from a vaccinated cow producing broadly neutralising antibodies to examine the structure of their HIV Env gp40 antigen binding sites and construct chimeric bovine-human antibodies. Methods: A cow was vaccinated with HIV- AD8 Env gp40 trimers over 4 years and antigen-specific memory B-cells. HIV specific memory B cells were detected in ELISPOT assay and isolated by FACS single-cell sorting from PBMC. The bovine Ig heavy (H) and light (L) chain variable (V) regions were amplified from cdna from anti-cd2+ anti-igg+ gp40-pe-binding+ cells by nested PCR. Paired H and L chain expression vectors using human Ig constant regions were made and co-transfected into 293T cells. Chimeric bovine-human (BH) Ig in supernatant was screened for HIV- gp40 binding in ELISA. Results: The frequency of HIV specific memory B cells was.96% ± 0.3% of total memory B cells on a background of 0.24% in nonimmune PBMC. Ig from 6 of 30 matched chimeric BH H and L chain plasmid transfections displayed strong binding to HIV- AD8 gp40 Env trimers using direct ELISA, but not gp20 monomers or cleaved gp4. Two mabs bound gp40 by mass-spectrometry and western blotting. Analysis of CDRH3 of the anti-hiv antibodies had a CDRH3 containing Cys and aromatic residues and were 4-22 amino acids (average size of 8.8 ± 3.). In addition, the somatic mutation rate in CDRH3 compared to the DH3 germline gene was % (Average: 87.34± 2.5%). The V-region sequence aligned most strongly with 2F5 and 4E0 patient derived mabs. Conclusions: The bovine V-gene CDRH3 size and frequency of somatic mutations of the isolated bovine Ig-genes raised through vaccination were comparable with those for human patient s elite neutralizing antibodies. 86 HIV Research for Prevention 204 HIV R4P

187 Posters : Informed Consent P.0 Developing a Novel Approach to Assess Stakeholder Views on the Length of Consent forms for HIV Prevention Research Amy Corneli, Emily Namey, Monique Mueller, Ansley Lemons, Jeremy Sugarman 2 FHI 360, Durham, NC, United States, 2 Johns Hopkins University, Baltimore, MD, United States Wednesday, 29 October P.02 Regulated Flexibility in Adolescent Informed Consent Procedures: Ensuring Inclusion of the Most Vulnerable Populations in HIV Research Rachael C. Dellar, Kshama Haribhai, Fanelesibonge Ntombela, Silvia Maarschalk, Ayesha Kharsany, Quarraisha Abdool Karim,2 CAPRISA, Durban, South Africa, 2 Columbia University, Epidemiology, New York, South Africa Background: Potential research participants often do not understand information in consent forms (CFs). Long CFs can be a contributing factor, yet empirical evidence is limited on the information that should be included or that could be removed from CFs in HIV-related research and on the barriers to reducing CF length. Methods: We are exploring these issues from the perspectives of stakeholders affiliated with the HIV Prevention Trials Network (HPTN): participants, investigators and site staff, community and regulatory representatives, institutional officials, and members of institutional review boards. We reviewed data collection approaches that could ) allow stakeholders to identify CF text as essential or extraneous and 2) identify areas of agreement and divergence across the stakeholder groups. We also explored approaches that could allow stakeholders to learn the perspectives of other stakeholders and to confirm or refute comments attributed to them. Results: A novel, modified Delphi approach was developed for building stakeholder consensus on how to shorten CFs. The three-step process begins with stakeholders highlighting essential or extraneous sentences of an HPTN CF on a tablet device and describing barriers to reducing CF length during a face-to-face interview. Next, up to three follow-up, online surveys will be conducted among a sample of participants from each stakeholder group. During each survey, stakeholders view results from the previous stage; refine their selections of essential information after considering all stakeholder responses, particularly for areas of continued disagreement; and reflect upon the barriers previously identified. The Delphi process concludes with an on-line group interview in which stakeholders will give input on a shorter CF built from the results of the prior steps. Conclusions: Consensus among key stakeholders about information to keep and remove from HIV-related CFs is critical for reducing CF length. Innovative methods should facilitate this process. Background: Adolescent-focused HIV prevention trials are key to altering predicted epidemiologic trajectories for HIV in hyper-endemic settings such South Africa. However, obtaining informed consent for adolescent participation in clinical and behavioral trials in such settings presents a number of challenges which can result in exclusion of the most HIV-vulnerable adolescents from HIV research. Methods: We assessed the major challenges to obtaining informed consent from assenting high school students under 8 years, and the uptake of more flexible informed consent procedures designed to overcome these challenges, in the behavioural school-based adolescent trial CAPRISA 007 based in rural KwaZulu-Natal, South Africa. Results: A total of 2675 students were enrolled into CAPRISA 007 in 200, of whom 98 students (72.0%) were under 8 years of age. Major challenges in obtaining informed consent for assenting students under 8 years were the significant proportions of students who were from child-headed households (8.8%), had absent parents or guardians (.3%), or had parents or guardians who failed literacy and comprehension assessments (3.9%). An important pathway developed and approved in collaboration with University of KwaZulu- Natal s Biomedical Research Ethics Committee to overcome such challenges and to introduce regulated flexibility in consent procedures were community groups which were permitted to give proxy consent for student participation in the trial. These community groups assisted directly in the consent procedures for 24.0% (46/98) of students who might otherwise been excluded from participation. Conclusions: Community engagement and the development of flexible and adaptive informed consent procedures are critical for facilitating the inclusion of the most vulnerable populations in HIV prevention trials targeted to adolescents. 87

188 Posters Posters : Informed Consent P.03 P.04 Assessment of Understanding for Informed Consent in HIV Vaccine Trials Graham C. Lindegger, Michael Quayle 2, Catherine M. Slack, Sagri Singh 3, Sabrina Welsh 3, Pat Fast 3 University of KwaZulu Natal, HIV/AIDS Vaccine Ethics Group (HAVEG), Pietermaritzburg, South Africa, 2 University of KwaZulu Natal, Psychology, Pietermaritzburg, South Africa, 3 International AIDS Vaccine Initiative (IAVI), New York, NY, United States Background: Informed consent (IC) is accepted as an essential ethical requirement for clinical trials, including HIV vaccine trials (HVT). Authentic IC requires that potential participants adequately understand various trial concepts and the implications of trial participation before being accepted into trials. While assessment of understanding is usually done using forced choice questionnaires, there are many limitations to this method of assessment. This paper reports on a series of studies done to compare various methods of assessment of understanding, including true/false questionnaires, vignettes and narrative approaches. Methods: Three studies have been conducted to date. The methodology involved quantitatively comparing the levels of measured understanding on three assessment tools, viz. forced choice questionnaires, trial related vignettes and narratives of trial participation. The comparisons were conducted first on a group of potential HVT trial participants in South Africa. A second study was conducted using a hypothetical HVT trial on samples of potential trial participants in South Africa and Zambia. A subsequent study compared measured understanding on these tools on samples of potential participants in HVT in Uganda and South Africa. Results: Results of all studies show a significant difference between measured levels of understanding, with the forced choice questionnaire having the highest scores, arguably over-estimating levels of understanding, and open-ended measures the lowest scores. Conclusions: It is suggested that open-ended methods of assessment provide more realistic and appropriate measures of understanding, which should be incorporated in IC procedures for HVT. Reservations have been expressed about the cost and skill implications of using such methods, however, suggestions are made for how these measures might be cost-effectively incorporated into HVT. Determinants of Informed Consent Comprehension among Fisher Folk Cohort in HIV Vaccine Preparatory Studies in SW Uganda Elizabeth Mbabazi, Andrew Abaasa, Gershim Asiki, Ubaldo Bahemuka, Eugene Ruzagira, Margaret Nambooze, Cissy Lilian Nalubega, Mathew A. Price 2, Anatoli Kamali Medical Research Council/Uganda Virus Research Institute, Entebbe, Uganda, 2 International AIDS Vaccine Initiative, New York, NY, United States Background: Informed consent comprehension is mandatory for individuals decision to enrol and participate in clinical trials. Low literacy may be a barrier to consent. We report the relationship between volunteer literacy and comprehension of study information at entry into a fisher folk HIV vaccine preparedness cohort. Methods: Community meetings were conducted at fishing sites along Lake Victoria to sensitize individuals to participate in HIV vaccine preparedness study. Interested individuals were provided HIV counselling and testing through which HIV uninfected volunteers were identified and invited to a study clinic located about 40km away. At the study clinic, written information on the study objectives, risks, benefits and procedures was read aloud and explained by a nurse to each individual. Those unable to read were requested to identify an independent witness to confirm provision of accurate information. Assessment of understanding (AoU) of study information was then done by a different study staff using 8 questions requiring true or false responses. Literacy was defined as the individuals ability to read the study information in the local language and write their names. For the analysis scoring all 8 points correctly was considered as a pass. Results: Of 584 (60% men) with mean age 28 years enrolled between Jan 202 and Mar 204, 22 (2%) were not literate. A total of 508 (87%) passed the AoU at first attempt. This did not differ by: literacy status (87% for literate and not literate), gender (86% men vs. 89% women, p=0.28] or other socio-demographic characteristics (age, occupation religion, marital status, number of dependants, tribe and parity) of the individuals. Conclusions: Literacy status and other demographic factors may not affect individuals comprehension of the informed consent process at study onset in these communities provided the investigator reads and explains the study information in the language understood by the individuals. 88 HIV Research for Prevention 204 HIV R4P

189 Posters : Informed Consent Wednesday, 29 October P.05 Participants Models of HIV Vaccine Trialrelated Concepts: The Dilemma of Trust for the Informed Consent Process Clinton L. Rautenbach, Graham C. Lindegger, Catherine M. Slack, Peter A. Newman 2, Melissa Wallace 3 University of KwaZulu Natal, HIV/AIDS Vaccine Ethics Group (HAVEG), Pietermaritzburg, South Africa, 2 University of Toronto, Factor-Inwentash: Department of Social Work, Toronto, ON, Canada, 3 Desmond Tutu HIV Foundation, Socio-Behavioural and Adolescent Division Leader, Cape Town, South Africa Background: Informed consent (IC) is a dialogical process between researchers and potential participants, which should be founded upon a reciprocal research rapport. A bilateral, dialogical process should be rooted in an ethos of credibility and trust. This paper will explore enablers and inhibitors of trust within the IC process in HIV vaccine trials (HVTs) in South Africa. Methods: The study employed an open qualitative, process-oriented exploratory design, using Focus Group Discussions (FGDs) to elicit mental models or key representations of critical research-related concepts from key constituencies, including CAB representatives, educators, and site staff involved in consent processes at an HIV prevention trial research site. Four FGDs were conducted - each constituency was interviewed separately - with approximately eight participants per group. Transcribed FGDs were subjected to thematic analysis by three qualitative researchers. Results: Findings showed several barriers to credibility and trust around research, researchers, and key research-related concepts indicated by community members, which may complicate how concepts are understood, interpreted or believed. Study respondents reported and engaged in several strategies to build trust and develop research rapport - likely to be foundational to research participation. We found socioeconomic, political, gender, race and culture-based mental models that may compete with or, alternatively, complement explanations of concepts offered by site-staff (e.g. vaccine-induced seropositivity [VISP]). Conclusions: Authentic IC should involve a systematic exploration of these barriers and enablers to trust as part of the on-going IC process, to complement efforts to promote and assess understanding. We make a series of recommendations for stakeholders who wish to incorporate trust-building into their engagement and consent work to enhance partnering and decision-making for HIV prevention trials. 89

190 Posters Posters 2: Innate Immunity P2.0 P2.02 Protective Role OF DC-SIGNR in HIV- Pathogenesis Omkar Chaudhary, Sanjeev Kumar, Muzamil A. Makhdoomi, Manju Bala 2, Jasbir Singh 3, Anjali Hazarika 4, Rajesh Kumar 5, Kalpana Luthra All India Institute of Medical Sciences, Biochemistry, New Delhi, India, 2 Apex Regional STD Teaching, Training & Research Centre, Vardhman Mahavir Medical College & Safdarjung Hospital, Microbiology, New Delhi, India, 3 Kurukshetra University, Biochemistry, Kurukshetra, India, 4 All India Institute of Medical Sciences, Blood Transfusion Services, Cardiothoracic and Neurosciences Center, New Delhi, India, 5 Society for Promotion of Youth and Masses Center, New Delhi, India Background: Dendritic cells (DCs) capture HIV- from periphery via DC- SIGN/R receptors and transfer to CD4 + T cells. The DC-SIGNR is highly polymorphic and changes in its repeat regions have been suggested to influence HIV- disease progression. Methods: Blood from 230 seronegative healthy individuals, 200 injecting drug users and 230 patients infected with HIV- was collected. DC-SIGNR polymorphism was performed by Polymerase chain reaction. The distribution of peripheral blood DCs and their subset frequency were determined by flow cytometry. DC-SIGNR expression in Peripheral blood mononuclear cells was determined in HIV- antiretroviral naïve patients and healthy individuals. DCs were cultured from monocytes and infected with HIV- Indian clade C virus. The expression of DC-SIGNR on DCs was silenced using DC-SIGNR sirna and the effect of down regulation of this receptor on HIV- infectivity of DCs, Co-stimulatory and signaling molecules. Results: The frequency of heterozygous DC-SIGNR 7/5 genotype and allele 5 was significantly higher in injecting drug users compared to HIV- infected patients and was associated with high dendritic cells count, CD4 + T cells and low DC-SIGNR expression, viral load. The expression of DC-SIGNR was higher in HIV- infected patients and inversely correlated with CD4 + T cell count. DC-SIGNR transfected DCs, the expression of co-stimulatory molecules and P38 MAPK was significantly reduced. Silencing of DC-SIGNR expression in DCs followed by infection with HIV- clade C viruses demonstrated lower levels of p24. Conclusions: DC-SIGNR 7/5 genotype and allele 5 may have a protective role in HIV- infection. The silencing of DC-SIGNR expression decreases the gene expression of the co-stimulatory molecules that in turn may inhibit the DC-T cell interactions needed for progression of HIV- infections. A long-term goal of our study is to develop novel therapeutic approaches to prevent HIV- infection. Skin Antigen-presenting Cells and Inflammation for Tailored Immunity to HIV Vaccines Behazine Combadiere, Clement Levin, Olivia Bonduelle, Charles Nuttens, Mireille Centlivre, Helene Perrin, CUT HIVAC Consortium INSERM U 35 CIMI-Paris and UPMC, Immunity and Vaccination, Paris, France Background: The EU-funded Cutaneous and mucosal HIV vaccination (CUT HIVAC) project are proposing alternative routes for vaccinating against HIV. A considerable part of CUT HIVAC work has been dedicated to understanding the mechanisms triggered after HIV vaccination via the skin for the induction of mucosal immunity by activation of T follicular helper cells. Skin outer surface protects from pathogens invasion thanks to skin antigen-presenting cells, which scan their changing microenvironment based on molecular changes that need to be identified. Methods: Using human skin explants model for vaccination as well as mouse models, we will present our understanding of skin cellular and molecular networks processes and aims at designing novel therapeutic approaches. Results: Langerhans and dermal dendritic cells pay a major role in dictating the quality of the immune responses in the draining lymph nodes and in programming mucosal immunity. Skin APC (Langerhans cells, dermal dendritic cells) surrounding the hair duct use their dendrite extension to scan their constantly changing microenvironment and interact to other skin cells (keratinocytes, fibroblasts, skin immune cells and other antigen presenting cells). In addition, inflammatory cells collaborate with professional antigen-presenting cells in the induction of CD8 in the draining lymph node and CD8 memory in the bone marrow as well as mucosal immunity in the vagina. Our work in mice model also highlights the importance of skin epidermal DCs in eliciting a strong T FH and B cell responses. It provides insight in skin vaccination mechanisms to improve the expansion of the T FH for vaccine efficacy. Conclusions: Thus, skin routes of immunization allow to target specific population of antigen-presenting cells and solicit inflammatory cells that could tailored cellular and humoral immunity for a preventive HIV vaccine development. 90 HIV Research for Prevention 204 HIV R4P

191 Posters 2: Innate Immunity P2.03 Wednesday, 29 October P2.04 Protective Role of TLR3 Induced Human Beta Defensins during Acute HIV- Infection Bjorn Corleis, Antonella Lisanti, Christian Korner, Molly A. Amero, Eric S. Rosenberg 2, Todd M. Allen, Marcus Altfeld,3, Douglas S. Kwon Ragon Institute of MGH, MIT, and Harvard, Cambridge, MA, United States, 2 Massachusetts General Hospital, Division of Infectious Diseases, Boston, MA, United States, 3 Heinrich-Pette-Institut, Leibniz Institute for Experimental Virology, Hamburg, Germany Background: Human beta defensins (HBD) and 2 (HBD2) are antimicrobial peptides expressed by epithelial cells and immune cells in tissue and peripheral blood. They are part of the first line of innate immune defense against invading pathogens at mucosal sites. Although beta defensins have been shown to have antiviral activity in vitro, their expression during the course of HIV- infection in vivo has not been fully characterized. Methods: We investigated production and activity of HBD and HBD2 in intestinal mucosal biopsies and peripheral blood mononuclear cells (PBMCs) from HIV--uninfected individuals (n=0) and subjects with chronic untreated HIV (n=0), immunologically controlled HIV (n=3) and acute HIV (n=23) using quantitative PCR and viral inhibition assays. In vitro studies of the effect of Toll like receptor (TLR) signaling on defensin expression were conducted using TLR specific ligands, inhibitors and sirna technology. Results: HBD expression is highly upregulated (p< ) in PBMCs but not in intestinal biopsy sample from acute HIV patients. Monocytes are the main producers of HBD in PBMCs whereas epithelial cells are the main source in the intestinal tract. HBD is upregulated in monocytes during acute infection (Fiebig stages 2/3) but returns to basal levels in chronic infection. In vitro studies reveal that HIV induces HBD but not HBD2 transcription in monocytes. HIV-induced HBD expression is dependent on TLR3 but not TLR 4, 5, 7 or 9 activation and independent of interferon or tumor necrosis factor alpha signaling. Importantly, recombinant HBD blocks HIV replication in activated T cells and suggests a protective role for defensins during acute HIV infection. Conclusions: Our study shows that the antiviral peptide HBD is upregulated in monocytes during acute HIV infection in a TLR3 dependent manner. Activation of the TLR3 pathway and upregulation of beta defensins at sites of infection might be an important defense mechanism during HIV acquisition and is currently under investigation. Chronic Untreated HIV- Infection Is Associated with Increased Erythrocyte Apoptosis and Inflammatory Monocyte Erythrophagocytosis Richard H. Glashoff, Stanley Loots, Hayley Ipp Stellenbosch University, Medical Virology, Cape Town, South Africa Background: Chronic HIV- infection is characterized by inflammation and also by anaemia. Erythrocyte apoptosis (erythroptosis) may contribute to anaemia and may also drive production of phagocytic monocytes/macrophages. In this study we investigated erythroptosis in asymptomatic, untreated HIV- infection and the relationship between erythrocyte death and phagocytic monocyte activity. Methods: A total of 44 chronically HIV- infected individuals (CD4 count > 200) and 33 matched uninfected individuals were included. Erythrocytes were stained with annexin V for determining ex vivo levels of erythroptosis. Erythrocytes were also subjected to oxidative stress in vitro (5mM hydrogen peroxide), in the presence or absence of the antioxidant N-acetyl cysteine (NAC). Finally, erythrocytes were also stained with CFSE and then incubated with purified autologous monocytes to monitor phagocytosis. Results: HIV- infected individuals had reduced haemoglobin levels (2.9g/dL vs. 4.g/dL, p=0.07). This was mirrored in decreased RCC. Significantly higher erythrocyte annexin V expression was observed (3.4% vs. 0.4%, p=0.089). Annexin V expression increased in both groups when exposed to hydrogen peroxide. NAC significantly reduced the percentage of apoptotic RBCs in the control group (20±5.% to 5±4.4%, p=0.006), but not the HIV- group (8.3±5.0% to 6±5.2%, p=0.065). There was a significant increase in the percentage inflammatory monocytes (CD4+CD6+) in the HIV- group (8.3±3.5% vs. 5.3±3.8% p=0.0054). The expanded inflammatory monocytes were more erythrophagocytic than classical monocytes, and preferentially phagocytosed apoptotic erythrocytes (phagocytic index 2.9% vs. 5.2%, p=0.008). Conclusions: Significant reduction in haemoglobin levels in untreated chronic HIV- infection was associated with increased erythroptosis. Inflammatory monocytes displayed enhanced uptake of erythrocytes. Limiting inflammatory toxicities may counteract the development of anaemia and associated monocyte changes in HIV- infection. 9

192 Posters Posters 2: Innate Immunity P2.05 Dysfunctional Neutrophil Responses to SIV Infection P2.06 Preliminary Evaluation of Serpins as Potential Candidate Microbicides Tiffany Hensley-McBain, Laura E. Richert-Spuhler, Michael Koday, Jillian Gile, Brandon F. Keele 2, Jacob D. Estes 2, Nichole R. Klatt Washington National Primate Research Center, University of Washington, Department of Pharmaceutics, Seattle, WA, United States, 2 Frederick National Laboratory for Cancer Research, AIDS and Cancer Virus Program, SAIC-Frederick, Inc., Frederick, MD, United States Background: Recent studies indicate that individuals with low neutrophils (PMN) are at increased risk of HIV infection. Also, the RV44 vaccine trial implicated non-neutralizing antibodies and associated Fcmediated functions in vaccine-induced protection. These studies suggest that early innate antiviral functions and Fc-mediated functions of PMN may be important mediators of protection. Consequently, protective immunity may rely on kinetics of PMN mobilization, activation, and recruitment during acute HIV/SIV infection. Methods: We assessed kinetic changes in PMNs in peripheral and mucosal tissues during acute SIV infection in six rhesus macaques challenged i.r. with 00,000 TCID50 of SIVmac239X. Flow cytometry, CBC, and luminex were used to assess PMN and cytokine levels and related PMN functional markers. Samples were collected pre-siv and days 3, 7, 4, 2, 28, 42, and 63 post-siv. Results: We observed a significant decrease in systemic IL-7 (p=.033) and a trending decrease in G-CSF (p=.0625) early after SIV. Surprisingly, blood PMN concentrations steadily decreased after infection, and no significant increase of PMN was detected in gut tissues. Blood PMN numbers and rectal PMN percentages significantly correlated (p=.0032), and HLA-DR (p=.033), CD86 (p=.056), and FcγRI (p=.033) were significantly upregulated on PMN during acute SIV infection. Conclusions: In contrast to other acute viral infection models, PMNsupporting cytokines are decreased or not induced during acute SIV, potentially contributing to lack of PMN mobilization from the bone marrow and recruitment to the tissues. Further, blood and rectum PMN levels correlate post-siv, suggesting that blood PMN concentration may directly impact recruitment to gut tissues. Lastly, upregulation of markers involved in antigen presentation and Fc-mediated functions highlight the potential diverse functional roles of neutrophils during acute SIV infection and the potential importance of inducing neutrophils for effective prevention strategies. Carolina Herrera, Natalia Olejniczak, Frank Plummer 2, Robin Shattock, Adam Burgener 2 Imperial College, Infectious Diseases, London, United Kingdom, 2 University of Manitoba, Winnipeg, MB, Canada Background: Studies on HIV-resistant women (exposed uninfected) from the Punwami Sex Worker cohort have shed light on putative protective mechanisms, suggesting that mucosal immunological factors, such as serpins, could be mediating HIV-resistance. This project aims to assess the activity of a panel of serpins against HIV- in a preclinical mucosal tissue explant model. Methods: Antiviral efficacy of nine blinded serpins was assessed in TZMbl cells, ecto-cervical tissue explants and migratory cells. Incubation of cells or tissue with serpins for h was followed by addition of an R5- tropic virus, BaL. Tissue was exposed to virus for 2 h and then washed. Following overnight incubation of tissue, migratory cells were harvested and co-cultured with PM- CD4+ T cells without drug. Infection was determined by measurement of luciferase expression (in TZM-bl cells) or p24 viral antigen in culture supernatants. Results: Following screening in TZM-bl cells where dose-response curves were measured for two serpins, antiviral activity in ecto-cervical explants was detected for five serpins allowing us to establish an order of inhibitory potency when mimicking pre-coital use of a serpin-based microbicide. In addition, the migration of cells out of the explants was blocked by certain serpins, indicating potential prevention of viral dissemination following amplification of the founder population. Conclusions: These results constitute the base for further development of these mucosal proteins as microbicides. Serpin combinations and sustained exposure of explants to serpins mimicking repeated dosing strategies will inform the dosing and formulation of these antiproteases which have shown potential as effective microbicides able to inhibit HIV- transmission in pre-clinical assays. 92 HIV Research for Prevention 204 HIV R4P

193 Posters 2: Innate Immunity P2.07 Wednesday, 29 October P2.08 Restoration of the NK Cells Ability to Mediate ADCC in HIV- Positives after Six Months of HAART Can be Explained by Normalization of their Phenotype Sanne Skov Jensen,2, Hans Jakob Hartling 3, Jeanette Linnea Tingstedt, Tine Kochendorf Larsen, Susanne Dam Nielsen 3, Court Pedersen 2, Anders Fomsgaard,2, Ingrid Karlsson Statens Serum Institut, Department of Microbial Diagnostic and Virology, Copenhagen, Denmark, 2 Odense University Hospital, Department of Infectious Diseases, Odense, Denmark, 3 Copenhagen University Hospital, Department of Infectious Diseases, Copenhagen, Denmark Skin Migratory APCs Fine-tune Lymph Node Microenvironment for the Generation of T Follicular Helper Cells and Mucosal Immunity Clement Levin,2, Charles Nuttens,2, Olivia Bonduelle,2, Helene Perrin,2, Bernard Verrier 3, Nicolas Fazilleau 4, Behazine Combadiere,2 Sorbonne Universités, UPMC Univ Paris 06, Unité Mixte de Recherche de Santé (UMR S) CR7, Centre d Immunologie et de Maladies Infectieuses, Paris, France, 2 INSERM U 35 CIMI, Paris, France, 3 Institut de Biologie et Chimie des Protéines UMR 5305 CNRS/UCBL, Lyon, France, 4 Centre de Physiopathologie de Toulouse-Purpan, UMR 043, CHU Purpan, Toulouse, France Background: Natural killer (NK) cell phenotype and function have recently gained much attention as playing crucial roles in antibodydependent cellular cytotoxicity (ADCC). Within the context of HIV infection, the ability of NK cells to mediate ADCC is critical for protection from disease acquisition and progression. Methods: We investigated NK cell function, as measured by ADCC, in HIV- positive individuals before and six months after highly active antiretroviral therapy (HAART) initiation. The ability of antibodies and NK cells to mediate ADCC was investigated separately and in combination in an autologous model. The NK cell subset distribution and NK cell phenotype were analyzed. The ADCC-GranToxiLux assay was used to evaluate the ability of NK cells and antibodies to mediate ADCC. The frequency of NK cells expressing receptors was determined by phenotypic labeling followed by flow cytometry. Results: The ability of NK cells to mediate ADCC was significantly increased after only six months of HAART and was not explained by a normalization of NK cell subsets but rather by a normalization of the NK cell phenotype. The frequency of NK cells expressing CCR7 and CD27 significantly decreased after six months of HAART. For individuals with no increase in ADCC after six months of HAART, the frequency of NK cells expressing NKp46 was down-regulated. The ability of antibodies to mediate ADCC alone and in combination in an autologous setup was not improved. Conclusions: HAART improves the ability of NK cells to mediate ADCC after six months. This improvement does not correlate with general immune restoration, as measured by CD4 + T cell counts, but rather to a normalization of the NK cell phenotype. The investigation of the immune function during HAART is important in order to gain knowledge about who may respond to a therapeutic vaccine. Background: The understanding of mechanisms of induction of humoral responses need to be further studied to define vaccination strategies against HIV. We previously showed that intradermal (i.d) immunization of mice with PLA-HIV-P24 nanoparticles induced mucosal immunity in the vagina. The skin is rich in specialized dendritic cells (DCs) subsets such as Langerhans cells (LCs) and dermal DCs, as well as other myeloid DC populations. We therefore questioned the role of skin antigen-presenting cells in the induction of T follicular helper cells (T FH ), which play a pivotal role in B cell help and isotypic class switch for shaping IgG and IgA-secreting B cells. Methods: We used Langerin-DTR mice model or ear ablation after ear i.d vaccination to study the role of skin cells and Langerin+ cells in T FH and IgG and IgA-secreting B cells induction. We used fluorescent PLA to track antigen uptake and migration by skin resident and inflammatory cells. Finally, gene array analysis was performed to reveal key markers of DLN inflammation after immunization. Results: Conditional depletion of Langerin+ cells or ear ablation resulted in partial to full abortion of T FH polarization and p24-specific IgG and IgA-secreting B cells expansion, respectively. Tracking of injected PLA nanoparticles showed high ability of LCs and Langerin+ dermal DCs to capture antigen in the skin and migrate to the T/B interface of DLNs. Gene array analysis of the whole DLNs of control and ear ablated mice after i.d immunization revealed distinct molecular signatures of inflammatory molecules critical for generation of immune responses. Conclusions: Work is currently in progress to validate the signature of innate immunity brought by skin APCs to the lymph nodes for T FH induction and mucosal immunity. Our work emphasizes on the role of migratory DCs from the skin in fine-tuning the DLN microenvironment. It will provide insights into mechanisms by which i.d immunization can induce superior humoral immune responses for the prevention of HIV infection. 93

194 Posters Posters 2: Innate Immunity P2.09 Acute SIV Infection Rapidly Depletes CD4 + pdcs, but Induces Gut-Homing pdcs from Bone Marrow Haiying Li, Tristan I. Evans, Michelle Connole, Jacqueline Gillis, Fay E. Wong, Yi Yu, Roger Keith Reeves New England Primate Research Center, Harvard Medical School, Southborough, MA, United States P2.0 Stimulation of Toll Like Receptor 7 and 8 (TLR7/8) Inhibits HIV- Replication in the Pregnant Uterine Mucosa Hicham El Costa, Heloise Quillay,2, Marion Duriez, Claire De Truchis 3, Anne Le Breton 3, Mona Rahmati 4, Julien Ighil 4, Françoise Barré-Sinoussi, Marie-Thérèse Nugeyre, Elisabeth Menu Institut Pasteur, Paris, France, 2 Univ. Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France, 3 A. Béclère Hospital, AP-HP, Clamart, France, 4 Pitié Salpétrière Hospital AP-HP, Paris, France Background: Given their potent antiviral functional role, plasmacytoid dendritic cells (pdcs), the major IFN-α-producing cells, remain a target of interest for HIV/SIV disease. pdcs accumulate in the gastrointestinal mucosae during chronic disease, but the kinetics of mobilization and trafficking are poorly understood. Furthermore, pdcs in multiple tissues show evidence of dysfunction through unclear mechanisms. Methods: This planned sacrifice study included 2 rhesus macaques six naïve and six intravenously infected with SIVmac239. Animals had colorectal biopsies prior to infection, followed by longitudinal blood draws, and tissue collection at day 4 post infection. Phenotypic and functional analyses were performed using polychromatic flow cytometry. Results: During acute SIV infection pdcs were rapidly depleted from blood, spleen and liver, but accumulated in the gastrointestinal tract. The gut-homing receptor, a4β7, increased significantly on circulating and splenic pdcs by day 4 following infection, suggesting that trafficking was a primary mechanism for the loss of pdcs from blood and lymphoid organs. Interestingly, compared with naïve controls, bone marrow pdcs in acutely infected rhesus macaques had increased proliferation and significantly higher levels of a4β7 expression, suggesting that mobilization and gut-homing of pdcs may be imprinted in the bone marrow. In multiple tissues CD4 + pdcs were identified as the major producers of IFN-α and TNF-α production compared to CD4 - pdcs, but were specifically lost during acute SIV infection. It was not, however, apparent whether loss of CD4 + pdcs was associated with specific depletion or downmodulation of CD4. Conclusions: We demonstrate SIV rapidly induces mobilization of bone marrow and circulating pdcs to traffic to the gut mucosae, but impairs the pdc functional response by depleting the CD4 + cytokine-producing subpopulation. Background: Elucidating the mechanisms underlying the natural control of HIV- mother to child transmission during the first trimester of pregnancy may help determine biologic correlates of protection against HIV- transmission in human mucosa. The decidua basalis (uterine mucosa) is the main materno-fetal interface. Decidual macrophages (dm) and natural killer (dnk) express functionnal TLR7/8. Since HIV- ssrna encodes for TLR 7/8 ligands that can mediate direct activation of the immune system, we investigated the potent role of TLR7/8 in the control of HIV- infection in the decidua. Methods: Deciduas were obtained from HIV- negative women undergoing elective abortions (8-2 weeks of amenorrhea). Decidual explants or purified dm were stimulated with R848 (TLR7/8 ligand). The culture supernatants were collected for soluble factor quantification. The explants and dm were then used for flow cytometry experiments or infected with R5 HIV- or HIV-/VSV-G-luciferase pseudotype. HIV- entry was assessed by supernatant transfer experiment. Postentry steps were investigated by PCR quantification. The role of R848 on the dnk cytotoxic potential was measured by flow cytometry. Results: HIV- replication was blocked efficiently in decidual explants or purified dm when R848 was added prior to infection and to a lesser extent when added several days after infection. Stimulated explants and dm produced high levels of MIP-α, MIP-β and Rantes and decreased significantly the expression of HIV- coreceptors. HIV- entry was inhibited by decidual soluble factors. The number of proviral copies was decreased in stimulated dm. TLR7/8 stimulated dm activated indirectly dnk cytotoxicity. Conclusions: Our findings provide evidence that TLR7/8 exerts a durable and long lasting anti-hiv- effect by targeting several steps of HIV- replication cycle and by activating the immune system.these findings could provide strategies for blocking HIV- infection of mucosa. 94 HIV Research for Prevention 204 HIV R4P

195 Posters 2: Innate Immunity P2. Wednesday, 29 October P2.2 Phenotypes of Monocytes and Monocyte Derived Dendritic Cells (MODC) in Antiretroviral Naïve Chronically Infected HIV Patients Nadesh Ngechae Nji, Jules C. Tchadji, Achile Nangue Nangue, Georgia Ambada, Carol Stephanie Ngane Sake, Abel Lissom, Edith Temgoua 2, Betrand Sagnia, Samuel Sosso 3, Rachel Kamgaing 3, Jule Clement Assob Nguedia Assob 4, Godwin Wapimawah Nchinda Chantal Biya International Reference Center for Research on the Prevention and Management of HIV/AIDS, Microbiology and Immunology Laboratory, Yaounde, Cameroon, 2 Chantal Biya International Reference Center for Research on the Prevention and Management of HIV/AIDS, Infirmary, Yaounde, Cameroon, 3 Chantal Biya International Reference Center for Research on the Prevention and Management of HIV/AIDS, Medical Analysis Laboratory, Yaounde, Cameroon, 4 Faculty of Health Sciences, University of Buea, Medcine, Buea, Cameroon Background: Monocytes are immune cells which differentiate into antigen presenting cells such as dendritic cells (DC). Monocytes have been sub divided into three subclasses, based on CD4 and CD6 surface markers; namely classical (CD4++CD6+), intermediate (CD4++CD6+) and non-classical (CD4+CD6+, CD4+CD6+) monocytes. There is a variation in the phenotypes of these subpopulations in HIV- chronically. Monocytes, are used to obtain MoDC in vitro, and in vivo for several immunological studies. It is thus imperative to verify if the variation between the different subclasses of monocytes can impact their ability to be converted to MoDCs. Methods: PBMCs purified from peripheral blood of participants, were used for surface staining with fluorochrome-conjugated antibodies characteristic of monocytes; and also to enrich for monocytes using CD4 magnetic beads and magnetic separation columns. The enriched monocytes were cultured in MoDC differentiation medium to obtain immature MoDCs, in which maturation signal was added (polyiclc) and cultured. Surface staining using antibodies was done at each stage to determine the phenotypic characteristics of the cell populations. Results: A fourth class of monocytes based on their expression of CD4 and CD6, (CD4++CD6++), which had a direct correlation with CD4+ absolute counts and an indirect correlation with viral load was noticed. There was progressive down regulation of CD4 on the monocytes as they converted to MoDCs and up regulation of maturation markers of DCs (CD80, CD86, CDc, CDa and CD83). Conclusions: Despite a variation in the different monocyte populations observed between HIV infected people and HIV negative donors, the monocytes still convert to MoDC similarly. Monocytes in Chronic HIV- Infection: Putative Gut Homing Markers and their Relationship with Immune Activation Karmistha Poovan, Hayley Ipp 2, Richard H. Glashoff Stellenbosch University, Division of Medical Virology, Department of Pathology, Cape Town, South Africa, 2 Stellenbosch University, Division of Haematology, Department of Pathology, Cape Town, South Africa Background: The GALT is a site of rapid replication of HIV- during early infection resulting in GIT damage and microbial translocation into systemic circulation. This fuels on-going inflammation and immune activation, which can persist despite the use of ARVs. Monocytes are key role players in innate immunity, and due to GIT damage in HIV- infection, homing of monocytes to this site is envisaged as key in the pathogenesis process. Methods: Whole blood samples were collected from HIV + (n = 39) and HIV- (n=26) volunteers from clinics across Cape Town. Cells were stimulated with mg/µl LPS for 4 hours and then stained with antibodies prior to analysis of 8 colour panels on a BD FACS CANTO II. Results: There was a significant increase in the percentage of inflammatory monocytes (CD4+6+), in the HIV+ group. There was a higher overall monocyte expression of several chemokine receptors such as CCR5 (53.4% vs. 0.0%), CCR (6.47% vs. 2.94%), CCR7 (86% vs %) and CCR2 (52% vs. 3.7%). Other markers that were increased include TRAIL-R (74.52% vs %), CD62L (29.5% vs. 5.56%) and IL-0 (3.67% vs. 0.48%). Upon stimulation monocytes appeared to respond similarly in both study groups, but the HIV+ group did display unique and significant increases in IL-0 (2.53%), CD69 (.4%), CD6 (4.09%) and HLA-DR (6.75%). The inflammatory subset showed an increased expression of CD69 at baseline. Conclusions: In HIV+ the total monocyte population shows a trend of increased migratory markers including gut specific markers CCR7 and CCR2, indicating they are primed to exit circulation to travel towards sites of infection. The increased inflammatory subset seen in HIV+ individuals is known to be associated with the persistence of the inflammatory state and with the release of pro-inflammatory cytokines. Although chronic HIV- infection does not seem to functionally impair monocytes, it does lead to up-regulated expression of key activation and homing markers, providing potential intervention biomarker targets. 95

196 Posters Posters 2: Innate Immunity P2.3 P2.4 Activation of Toll-like Receptor 2 (TLR2) Heterodimers by HIV- Proteins Significantly Increases HIV Infection and Inflammation Bethany M. Henrick,2, Xiao-Dan Yao,2, Kenneth L. Rosenthal,2 McMaster University, Department of Pathology & Molecular Medicine, Hamilton, ON, Canada, 2 McMaster University, McMaster Immunology Research Centre, Hamilton, ON, Canada Background: Immune activation is a critical driver of HIV- infection and pathogenesis, however our understanding of HIV innate immune activation remains incomplete. Recently, we showed that soluble TLR2 directly interacted with HIV proteins. Here, we investigated TLR2 and its heterodimers as innate pattern recognition receptors for HIV- structural proteins. Methods: Using cells stably expressing TLR2, primary human T cells and cell lines expressing selected TLRs, we determined the ability of HIV structural proteins to activate IL-8 production through a NFκB signalling pathway and the TLR2-dependent effect on HIV infection. Knockdown of TLR expression with sirna and anti-tlr2 antibodies were used to confirm our results. Co-immunoprecipiatation was used to determine direct interaction of TLR2 with HIV proteins. Results: Cells expressing TLR2 had significantly increased HIV- integration and NFκB-dependent IL-8 production compared to cells lacking TLR2. Primary T cells exposed to TLR2 ligand, Pam3CSK 4, or HIV- p7, p24 or gp4, but not to gp20, produced significantly more IL-8 compared to cells incubated with anti-tlr2 antibodies. HIV p7 and gp4 were recognized by TLR2/, while p24 activated cells through TLR2/6. These results were confirmed by TLR2/ sirna knockdown, as well as studies of HEK293 cells expressing selected TLRs. HIV gp4, p7 and p24 were shown to co-immunoprecipitate with TLR2. Interestingly, in competition assays, p24 blocked p7 and gp4-induced TLR2 activation, thus providing a novel mechanism by which HIV can modulate innate sensing. Conclusions: Our results demonstrate that HIV- structural proteins p7, p24 and gp4 act as pathogen-associated molecular patterns (PAMPs) and signal through TLR2 and its heterodimers leading to significantly increased HIV infection,nfκb-dependent activation and proinflammatory cytokine production. These findings have important implications for our understanding of HIV immune activation, pathogenesis and vaccine development. Phenotypic Characterization of Natural Killer (NK) Cells in Antiretroviral Naïve Chronically Infected HIV- Patients (the CIRCB Afrodec cohort) Carole Stéphanie Sake Ngane,2, Jules C. Tchadji,3, Achille Nangue, Nadesh Nji, Georgia Ambada,3, Edith Temgoua, Samuel Sosso, Rachel Kamgaing, François-Xavier Etoa 2, Godwin Nchinda -CIRCB/Microbiology and Immunology Lab, Yaoundé, Cameroon, 2 University of Yaoundé I, Department of Biochemistry, Yaoundé, Cameroon, 3 University of Yaoundé I, Yaoundé, Cameroon Background: Natural Killer (NK) cells are effectors cells of the innate immune system which play a critical role as a first line of defense against viral infection.they are characterized as CD3-CD56+CD6+/- cells and effect cytotoxic activity against target cells as well as cytokines and chemokines production. NK cytotoxicity is modulated by inhibitory and activating receptors. HIV- infection maintains immune system in a sustained state of activation and causes immune dysfunction.however the impact of NK cell phenotype modulation has not been acessed in the context of Antiretroviral naïve HIV- infection. We shall verify the existence of HIV-specific NK cells in antiretroviral naïve chronically infected HIV- patients. Methods: 35 treatment naïvehiv- patients, CD4 greater than 350 cell/µl and 30 healthy donors were recruited. Age ranged from 2 to 65 years old. Patients group was composed of 05 patients with Viral load(vl)< 2Log,23 VL between 2-4,5 Log and 07 VL>4,5 Log.PBMCs were obtained from whole blood using ficoll-paque density grandient. NK cells are analyzed by multiparametric flowcytometry in bulk PBMCs. samples were aquired using BD FACScanto II machine and data analysed by flowjo 7.4 Results: There was a differential modulation of NK cell phenotype as we observed a decreased expression of activating markers NKP30, NKP44, NKP46 and a significant increase (P< 0,05) in the expression of HLADR, CD38 and NKG2D in treatment naïve HIV- infected patients relative to healthy donors. Down regulation of NKG2A expression was also observed in the patients group. Conclusions: This study shows phenotypic pertubations on NK cells in treatment naive HIV- infected patients. Observation of increased expression of activating receptors NKG2D, CD38 and HLA-DR is a pathological signature of HIV- infection however a decreased NKG2A in expression could be necessary in increase cytolytic function of NK cells. 96 HIV Research for Prevention 204 HIV R4P

197 Posters 2: Innate Immunity P2.5 Human Beta Defensins and RNases: Antiviral Effect during Sexual Exposure to HIV- Wednesday, 29 October P2.6 Sex Differences in TNFAIP3 Levels in pdc upon TLR7 Stimulation Wildeman Zapata,2, Wbeimar Aguilar-Jimenez, Zhimin Feng 3, Aaron Weinberg 3, Aniello Russo 4, Nicoletta Potenza 4, Hernando Estrada 5, Maria Teresa Rugeles Universidad de Antioquia UdeA, Facultad de Medicina, Grupo Inmunovirología, Medellin, Colombia, 2 Universidad Cooperativa de Colombia, Facultad de Medicina, Grupo Infettare, Medellin, Colombia, 3 Case Western Reserve University, School of Dental Medicine, Department of Biological Sciences, Cleveland, OH, United States, 4 Second University of Naples, Department of Life Sciences, Naples, Italy, 5 HERES Health, Lending Institution of Health, Santa Marta, Colombia Background: Individuals demonstrating natural resistance to HIV infection, despite repeated unprotected sexual exposure, are referred to as HIV--exposed seronegative (HESN), and are considered a model to determine mechanisms of protection. Innate immune factors, such as human beta defensins (HBD), and RNases, previously known for their antiviral potential, are present at mucosal barriers, main ports of HIV entry, suggesting their involvement in avoiding the establishment of HIV infection. Methods: To evaluate the role of these factors in the natural resistance to HIV-, we performed a study, including 60 HESN, and 6 healthy controls (HC). Vaginal, endocervical and oral mucosal samples were taken from these individuals; RNA was extracted, and mrna transcripts of HBD-2, -3, and the Ribonucleases, eosinophil-derived neurotoxin (EDN), RNase- and angiogenin were quantified by qpcr. In addition, we explored the step(s) of the viral cycle that was inhibited by these soluble factors, using an in vitro single-round, recombinant-based, viral infectivity assay. Results: Oral mucosa of HESN expressed significantly higher levels of hbd-2 (p=0.04) and hbd-3 (p=0.04) mrna compared to HC. Likewise, vaginal mucosa of HESN showed significantly higher mrna levels of RNase- compared to HC (p=0.036); in addition, endocervical mucosa showed higher mrna levels of EDN (p=0.04) and Angiogenin (p=0.02) compared to HC. Finally, HBDs and RNases inhibited HIV- replication, in a dose dependent manner at the following steps of the viral replication cycle: entry, reverse transcription and nuclear import. Conclusions: Taken together, these results suggest that innate immune factors, such as HBD-2 and -3, and RNases inhibit HIV- infection during either oral or genital exposure of HIV. In fact, HBDs and RNases inhibited in vitro replication of HIV-, pointing their potential role as antiretroviral molecules to be explored in the near future. Susanne M. Ziegler, Morgane Griesbeck 2,3, Judy Chang 4, Marcus Altfeld,2 Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Viral Immunology, Hamburg, Germany, 2 Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States, 3 CIMI/ U35, Paris, France, 4 Monash University, Department of Infectious Diseases, Melbourne, Australia Background: The outcomes of many diseases differ between women and men. It is well documented that women have a higher incidence as well as more severe pathogenesis of autoimmune diseases. Furthermore, accumulating evidence indicates a role of sex-based divergence of infectious diseases, including influenza and HIV-. In HIV- infection, clinical studies have shown faster disease progression and stronger immune activation in females compared to males for the same level of viral replication, as well as better control of initial viremia in women during primary infection (Meier et al 2009 Nat Med, Sterling et al 200 NEJM, Farzadegan et al 998 Lancet). It has been suggested that this is mainly due to Toll-like receptor (TLR)-mediated responses of plasmacytoid dendritic cells (pdcs), the main producers of IFN-α. We investigated the role of the TNF-α induced protein (TNFAIP) 3 (A20), an estrogen-regulated early NF-κB-responsive gene that encodes a ubiquitin-editing protein involved in negative feedback regulation of NFκB signaling. Thus, TNFAIP3 is known to be a potent anti-inflammatory signaling molecule that restricts multiple intracellular signaling cascades including TLR signaling. Methods: 00 pdc were sorted and stimulated for 2h with a TLR7 ligand. Using fluidigm technology the expression level of TNFAIP3 mrna was determined. Results: We detected a significant difference in TNFAIP3 mrna upregulation in pdcs after 2h TLR7 stimulation between males and females (p< 0.05). pdcs from males showed a 2-log higher expression level of TNFAIP3 mrna compared to females, associated with lower production of IFN-α in pdcs from males in response to HIV--derived TLR7 ligands and HIV-. Conclusions: Taken together, these data demonstrate that sex differences described in response to HIV- are associated with the enhanced expression of the sex hormone-dependent inhibitory regulator of TLR signaling TNFAIP3 in pdcs of men compared to women, providing a novel target to modulate the inflammatory IFN-α response to HIV

198 Posters Posters 2: Innate Immunity P2.7 Phenotypic and Functional Characteristics of Siglec-7, NKG2A and NKG2C Expressing NK Cells in Chronic HIV- Clade C Infection Michael Z. Zulu, Kewreshini Naidoo, Zenele Mncube, Manjeetha Jaggernath, Philip Goulder,2, Thumbi Ndung u,3, Marcus Altfeld,3,4, Christina Thobakgale Doris Duke Medical Research Institute, University of KwaZulu-Natal, HIV Pathogenesis Programme, Durban, South Africa, 2 Peter Medawar Building for Pathogen Research, University of Oxford, Department of Paediatrics, London, United Kingdom, 3 Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States, 4 Heinrich-Pette Institute, Department of Viral Immunology, Hamburg, Germany Background: Natural Killer (NK) cells play a critical role in the control of HIV- infection. HIV viremia has been shown to induce several phenotypic and functional abnormalities in NK cells such as a decrease in NKG2A+ and expansion of NKG2C+ NK cell population. Also, Siglec-7 was identified a cellular marker associated with NK cell abnormalities in HIV- infection. However, there is still a paucity of data on factors mediating NK cell receptor changes and identification of dysfunctional NK cell subsets during different stages of clade C HIV- infection. To assess mediators of NK cell dysfunction, our study examined the phenotype and function of NK cells based on Siglec-7, NKG2A, NKG2C and CD57 expression in HIV- clade C chronically infected individuals versus healthy donors. Methods: NK cell phenotypic profiles were characterized by assessing Siglec-7, NKG2A, NKG2C and CD57 expression on PBMCs of combination antiretroviral therapy (cart)-naïve HIV- chronically infected individuals (n=5), HIV- chronically infected individuals who have been on cart for at least 2 months (n=5) versus healthy individuals (n=5). The cytolytic potential of individual NK cell subsets between the three study groups was determined by flow cytometry assessment of CD07a and IFN-γ following overnight stimulation with K562 target cells. Results: Healthy individuals had the highest frequency of NK cells expressing Siglec-7 when compared to HIV-infected subjects (p=0.002). Interestingly, CD57 expression on total NK cell population was the highest in healthy donors compared to HIV- infected subjects (p=0.0). Overall, NKG2C expression in total NK cells was higher compared to NKG2A; however, there were no significant differences in NK cell degranulation and IFN-γ secretion across the three study groups. Conclusions: HIV- viremia alters Siglec-7 and CD57 expression on NK cells of infected subjects. Phenotypic changes induced by viremia may lead to functional dysregulation on NK cell subsets during chronic HIV- clade C infection. 98 HIV Research for Prevention 204 HIV R4P

199 Posters 3: Key Populations P3.0 Wednesday, 29 October P3.02 Awareness, Knowledge of HIV and Utilisation of HIV Preventive Services among Young People in Nigeria Mojisola Morenike Oluwasanu, Olatunji O. Adetokunboh 2 University of Ibadan, Health Promotion and Education, Faculty of Public Health, Ibadan, Nigeria, 2 Stellenbosch University, Division of Community Health, Cape Town, South Africa Background: Human Immunodeficiency Virus (HIV) infections remain a public health challenge in Nigeria. Despite the widespread knowledge of HIV coupled with the expanded access to HIV-related services, risky behavioural practices are prevalent among young people and the utilization of HIV preventive services remains low. The aim of this study is to examine the trend in the knowledge of HIV and utilisation of HIV preventive services among young people in Nigeria between 2003 and Methods: Data was obtained from Nigeria Demographic Health Survey conducted in 2003 and The data for indicators on comprehensive knowledge of HIV, comprehensive knowledge of prevention of motherto-child transmission, percentage who had ever had HIV test and collected result, percentage of pregnant women who were counselled, tested for HIV and received results and male circumcision for the 5-24 years age group were extracted. The statistical significance of the observed trend was tested. Results: In 2008, level of comprehensive knowledge of HIV among female youth was 22.2% while in 2003, it was 32.6%. Comprehensive knowledge of prevention of mother-to-child transmission among women increased from 5.% to 23.2% and from 4.4% to 29.3% among men in 2003 and 2008 respectively. In 2003, only 4.6% women had ever had HIV test and collected result and in 2008 this increased to 9.2% while among men, the percentage increased from 6.4% to 7.4%. Only 45.4% of the women interviewed knew where to get tested for HIV in contrast to 59.3% men. In 2008, only 8.4% pregnant women were counseled, tested for HIV and received results and 97.7% male has had circumcision done. Conclusions: The level of awareness and utilisation of HIV preventive services is very low in Nigeria among the youth. Male circumcision is an exceptional case because it is a culturally and religious norm for almost every male to be circumcised in Nigeria. There is a need to change the strategy for HIV youth awareness and utilisation of HIV services in Nigeria. Prior HIV Testing and ARV Use among HIV Positive Female Sex Workers in Kigali Rwanda Nurilign Ahmed, Etienne Karita, Rachel Parker 2, Rosine Ingabire, Julian Nyombayire, Robertine Sinabamenye, Jean Nduwamungu 3, Jean Bizimana 3, Gisele Umviligihozo 4, Honoree Uwamahoro 3, Amanda Tichacek 2, Eric Hunter 2, Susan Allen 2 Project San Francisco, Kigali, Rwanda, 2 Emory University, School of Medicine, Department of Pathology and Laboratory Medicine, Atlanta, GA, United States, 3 Projet San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 4 Profectus Biosciences, Kigali, Rwanda Background: In Rwanda, an estimated 3% of the adult population is infected with HIV (3.6% of adult women and 2.3% of adult men). This prevalence is higher in urban setting 7.% compared to rural setting 2.3%, and HIV risk is highest in urban FSW. Urgent access to ART for FSWs is vital to reduce new infections in clients. Rwanda national guidelines for ART for FSWs affirm that HIV+ FSWs should initiate ART regardless of CD4 cell count. Methods: FSWs were invited for services at Project San Francisco in Kigali Rwanda from September 202- April 204. Prior HIV testing, use of ARVs, and risk factors including average number of clients, and disclosure of HIV status to/from clients were assessed. Results: In this period, 677 FSWs were invited and tested for HIV and 372 (55%) were HIV+. Of these, 265 (7%) knew their HIV status beforehand and 86 (70%) FSWs were using ARVs. On average, FSW had 6 clients/month, one fifth of whom were regular/repeat clients. 5% of HIV+ FSWs disclosed their HIV status to their clients (73% willingly, 27% by client request). 25% of HIV+ FSWs reported requesting the HIV status of their clients and % of HIV+ FSWs were willing to get tested with their clients. FSW charged a median of $8 for services (range $2-$50) and reported a variety of successful strategies to minimize risk of violence from clients. Oral sex was reported by 0% and anal sex by 5% of FSW; 3% reported using condoms all the time and 44% most of the time. One in three were illiterate in the local language and one in eight could understand French or English. Conclusions: HIV testing should be promoted among FSW and counseling should emphasize Rwanda guidelines regarding ART. Research is needed to better understand obstacles to testing and ART in HIV+ FSW. It is encouraging that many HIV+ FSW disclose their HIV status to clients and are willing to be jointly tested with regular/repeat clients; this may be an avenue for prevention research. Inconsistent condom use remains a concern. 99

200 Posters Posters 3: Key Populations P3.03 Knowing Whom we Are trying to Protect: An Assessment of HIV Risk in South African Adolescent Females P3.04 Expanded Use of Antiretrovirals (ART) for Treatment and Prevention for Female Sex Workers in South Africa Shaun L. Barnabas,2, Heather B. Jaspan 3,4, Smritee Dabee, Shameem Z. Jaumdally, Hoyam Gamieldien, David Lewis 5, Anna-Lise Williamson,6, Thola Bennie 2, Angel Phuti 2, Martin van der Watt 7, Janan Dietrich 7, Nicola Mulder 8, Clive Gray 9, Thomas J. Hope 0, Francesca Chiodi, Robin Shattock 2, Lynn Morris 5,3, Nonhlanhla N. Mkhize 5, Glenda Gray 7,4, Linda-Gail Bekker 2,5, Jo- Ann S. Passmore,6, Women s Initiative in Sexual Health (WISH) University of Cape Town, Medical Virology, Cape Town, South Africa, 2 Desmond Tutu HIV Foundation, Cape Town, South Africa, 3 University of Cape Town, Department of Immunology, Cape Town, South Africa, 4 Seattle Biomedical Research Institute, Seattle, WA, United States, 5 National Institute for Communicable Diseases, Johannesburg, South Africa, 6 National Health Laboratory Service, Cape Town, South Africa, 7 Perinatal and HIV Research Unit, Johannesburg, South Africa, 8 University of Cape Town, Computational Biology, Cape Town, South Africa, 9 University of Cape Town, Immunology, Cape Town, South Africa, 0 Northwestern University, Department of Cell and Molecular Biology, Feinberg School of Medicine, Chicago, IL, United States, Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 2 Imperial College, Department of Infectious Diseases, Division of Medicine, London, United Kingdom, 3 National Health Laboratory Service, Johannesburg, South Africa, 4 HIV Vaccine Trials Network, Johannesburg, South Africa, 5 Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa Background: South African adolescent females have been underrepresented in HIV prevention trials though they have a disproportionately high risk for HIV acquisition. We hypothesized that changes associated with puberty may influence this susceptibility. Methods: The Women s Initiative in Sexual Health (WISH) study is investigating factors associated with HIV risk in young South African women, by enrolling HIV-negative adolescents between 6-22 years. This study plans to enroll 50 participants from Masiphumelele, Cape Town and 00 from Soweto, Johannesburg. As part of WISH, information on vaginal ph and the prevalence of STIs (C. trachomatis, N. gonorrhoea, T. vaginalis, M. genitalium, HSV-2, syphilis, candida) and bacterial vaginosis (BV) was collected. Participants were surveyed on sexual risk and hormonal contraceptive use. Results: Preliminary results in 48 enrolled participants show the majority of adolescents used injectable hormone contraception (80% Nur-Isterate and 6% Depo-Provera), with only 4% choosing oral contraceptives. Almost all, 98%, of the participants self-identified as heterosexual. Their median age of sexual debut was reported to be 6 years (IQR 6-7); with their median lifetime number of sexual partners being 2 (range -5). Reported condom use with their last sex act was 70%. Of the participants, 20% had been pregnant and all carried to term. Although only 25% reported having had a previous symptomatic STI, approximately half of the women were infected with an STI at enrolment; with C. trachomatis being the most common by far. Furthermore, 40% had BV (Nugent >7), with a median vaginal ph of 4.9. While STI prevalence at follow-up visits declined following treatment, BV did not. Conclusions: These data highlight the vulnerability of South African adolescent females to STIs, BV and potentially HIV despite self reported condom use. This study will provide insight into the reproductive health of South African adolescents and may influence the design of future preventative strategies. Robyn Eakle,2, Gabriela Gomez 3, W.D. Francois Venter, Helen Rees Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 2 London School of Hygiene & Tropical Medicine, Infectious Disease Epidemiology, London, United Kingdom, 3 Amsterdam Institute for Global Health and Development, Amsterdam, Netherlands Background: Recent trials demonstrated the efficacy of ART in reducing the risk of transmission in HIV-positive patients and of pre-exposure prophylaxis (PrEP) in reducing the risk of HIV acquisition. Adherence and implementation challenges relating to service delivery may hinder the ability of these interventions to make a significant impact. Sex workers have been identified as a critical community to access HIV prevention technologies. Methods: This demonstration project seeks to test the real-world viability of offering oral PrEP (for those testing HIV negative) and immediate HIV treatment (for those testing HIV positive) to female sex workers (FSWs) to prevent acquisition and transmission as part of a routine package of clinical care in two urban sites in South Africa. The main outcome studied will be retention at 2 months. The study will have two arms: ) PrEP arm for those testing negative; and 2) immediate treatment arm where women will be started on treatment for CD4 counts of >350 cells/mm 3. We will investigate process and other health indicators (i.e. uptake, safety, rates of pregnancy, sexual behaviour, rates of co-infections, and use of SMS reminders). A qualitative research component will aim to better understand the motivations and barriers to uptake of PrEP and immediate treatment from perspectives of participants and providers. Finally, an economic evaluation will inform a cost-effectiveness analysis combined with estimates of impact through epidemiological modelling. Results: In early 207, after following FSWs for up to 24 months, we will have finalised the analyses of all outcomes, as well as clinical data, qualitative research, cost data, and impact. Conclusions: This is one of two demonstration projects integrating immediate treatment and PrEP service delivery in a real world setting. Our results will aim to inform national and global policy making of the viability of PrEP and immediate treatment as interventions prioritised for high risk populations. 200 HIV Research for Prevention 204 HIV R4P

201 Posters 3: Key Populations P3.05 Wednesday, 29 October P3.06 Tenofovir Gel Use in Women at High Risk of HIV Infection: A Retrospective Analysis of the Sex Worker Sub-group within the CAPRISA 004 Cohort Tanuja N. Gengiah, Lise Werner, Quarraisha Abdool Karim,2, Salim S. Abdool Karim,2 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa, 2 Columbia University Mailman School of Public Health, New York, NY, United States Facing our Fears : Facilitated Film Viewings as a Community Engagement Tool in Research Involving MSM in Kenya Evans Gichuru, Salla Sariola 2, Elisabeth M. Van der Elst, Peter Mugo, Murugi Micheni, Susan M. Graham,3, Catherine Molyneux,2, Eduard J. Sanders,2 KEMRI-Wellcome Trust Research Programme, Kilifi, Kenya, 2 University of Oxford, Headington, United Kingdom, 3 University of Washington, Seattle, WA, United States Background: Sex workers in Durban, South Africa are at high risk for HIV infection and would benefit from effective prevention technologies. A small urban sex worker cohort participated in the CAPRISA 004 trial and their demographics, trial outcomes and adherence estimates have not been exclusively described previously. Methods: We conducted a retrospective analysis of demographic data, sexual frequency, gel applicator use and assessed adherence using applicator returns in conjunction with behavioural data from the CAPRISA 004 trial sex worker cohort. Results: In total 8 eligible female sex workers (self-identified) were enrolled (two ineligible women were excluded from this analysis). At baseline median (IQR) age was 22 years (IQR: 20-27) and 27.8% completed high school. At enrolment, approximately 77.8% reported condom use at last sex act, and 6.% reported having a stable partner. Eleven participants were randomized to the placebo arm and 7 to the active arm. Overall, median trial participation duration was 2 months (IQR: -28 months). Median sex acts reported per month was 0 (IQR: 7 to 3.5) and median number of applicators dispensed per month was 3 (IQR: 0-20). Of these 60% (IQR: 40-93%) of applicators were returned as used. Adherence, based on returned used applicators and self -reported sex, was 50% (IQR: 45-00) overall. HIV incidence was 7.53 per 00PY (95% CI ). However, there were only two HIV infections, both being in the placebo arm and HIV incidence in that arm was 3.8 per 00PY (95%CI ). This is in contrast to the HIV incidence rate in the parent trial which was 9. per00py (95%CI ) in the placebo arm. Conclusions: Despite their high risk for HIV infection the small CAPRISA 004 sex worker cohort demonstrated moderate adherence to study gel. However, it is noteworthy that the active arm had 00% protection and that the sex workers had higher adherence to this coital gel regimen compared to women prescribed daily gel in the VOICE trial in South Africa Background: Research with men who have sex with men (MSM) in Africa requires ongoing community engagement to ensure safety and access to care of participants. We assessed views of stakeholders regarding a community engagement film more than 3 years after a research clinic was attacked in Coastal Kenya. Methods: We made a 6-minute video Facing Our Fears capturing footage of the attack in 200, interviews with attack leaders and victims in 200 and 203, and various reflections of community members on MSM research. We conducted 0 facilitated film viewings (FFV) followed by focus group discussions (FGD) with 8-20 participants each, comprising of religious leaders, LGBT members, health care providers, policy makers and media representatives to assess the film s usefulness as a communication tool, perceived security risks if the film would be released publicly, and if it supported community ownership of MSM research. FGD were tape-recorded or noted by hand. Transcripts were analysed thematically by two of the authors. Results: FFV presented a platform for discussion about involvement of MSM in medical research, importance of stakeholder collaboration, and LGBT rights in general. Visual representations of community engagement efforts were seen to be effective in highlighting the importance of including MSM in health care and research. FGD participants identified concerns over the possible risks to LGTB communities and those working with them following public release, due to homophobic and discriminatory atmosphere in much of Africa. Conclusions: FFV were seen as an empowering tool for raising awareness of MSM research and same-sex relations in general. Facilitated viewings allowed the film to be used as a training tool for successful LGBT research with community engagement, and may mitigate the possible risks of further stigma following open release. 20

202 Posters Posters 3: Key Populations P3.07 Are Nigerian Healthcare Providers (HCP) Prepared for Men who Have Sex with Men (MSM): Lessons from the Mystery Client Survey in Nigeria? Chiedu Chike Ifekandu,2 Population Council, HIV Division, Abuja, Nigeria, 2 Ahmadu Bello University, Development Communications/Theatre Arts, Samaru-Zaria, Nigeria P3.08 The Anti-same Sex Marriage Law Implications on HIV Interventions for Men who Have Sex with Men in Nigeria Chiedu Chike Ifekandu, Bala Abullahi, Ibrahim Suleiman, Hafiz Abdullahi, Jean Njab Population Council, HIV Division, Abuja, Nigeria Background: Services in the most healthcare facilities in Nigeria are targeted at the general population; the services they provide tend not to be friendly towards the health needs of men who have sex with men (MSM). The need to address the situation informed the mystery client survey to health facilities to evaluate the quality of treatment, care and services rendered to MSM in Nigeria. Objectives: To evaluate the quality of HIV prevention services for Men who have sex with men in Nigeria. Methods: The study was conducted in 2 Nigerian states. Mystery Client Survey methodology was used for the study. Pre-recruited MSM visited healthcare Facilities anonymously, posing as a regular customer/client. After leaving the facility, a standardized reporting form (questionnaire) was discretely completed by the mystery client immediately after each visit to capture the facility with regard to their assessment of provision of quality services. The mystery client in this survey sought one of the two services: HCT or STI consultation services. Quantitative data was entered and analyzed using SPSS version 20.0 Descriptive statistics, including 95% confidence intervals (CI) and Chi square tests of comparison for differences between categorical variables were conducted. Results: The mean age of the respondents is 22years +/-SD. The study size is 04 MSM. More than 76% reported that information at the clinic was neither inclusive of their specific sexual needs nor address stigma against MSM. About 40% reported that their sexual practices and behaviours were not explored during the pre and post-test counselling. More than 50% reported that the HCP exhibited a judgmental attitude towards MSM, while 47% reported that follow-up, care and support as well as referral were not made after the pre and post-test counselling sessions. Conclusions: From this study, sensitizing HCP on sexual diversity can help in changing their attitudes and believe towards. Background: HIV prevalence in Nigeria among men who have sex with other men is at 7.2% (IBBSS 200). Despite the alarming prevalence figures, Nigerian president signed the anti-same-sex bill into an Act. This law does not criminalize MSM alone, but incurs 0 years jail sentence for institutions and healthcare providers working with MSM as well. Objectives: To evaluate how the law will impact on HIV interventions and possibly come up with a more effective strategy for field implementation. Methods: There are 48 MSM peer educators on the Strengthening HIV prevention Services (SHiPS) project in four northern Nigerian states. These 48 peer educators reach a total of 200 MSM with six month timeline. The strategies being utilized before the Act was enacted was the combined prevention packaging of intervention (CMPPI); using the cohort formation approach. A well-structured questionnaire were administered to the 48 peer educators on their background knowledge of the MSM community within their area of coverage, their knowledge of the content of the antisame-sex Act and the impact on peer participation in the SHiPS Project and brainstorm on new strategies in reaching the MSM peers without exposing them to danger. Results: More than one-quarter of the peer educators reported than most of their peers are declining participation for SHiPS project. Over half of the peer educator have read the same-sex marriage prohibition Act and believes that they are vulnerable to harassments from the law enforcement agencies or the general public while having their peer sessions. Almost all the peer educators suggested on the change of the approach from peer education sessions to inter-personal Communication (IPC). Conclusions: Advocacy led the National Agency for the control of AIDS should be paid to the federal ministry of health and the presidency using informed evidence from national survey to convince the government from public health angle while MSM should not be criminalized. 202 HIV Research for Prevention 204 HIV R4P

203 Posters 3: Key Populations P3.09 Wednesday, 29 October P3.0 Need to Raise the Level of Knowledge of PrEP and Rectal Microbicides (RM) among Men who Have Sex with Men (MSM) HIV and Sexually Transmitted Infection (STI) Testing among Female Sex Workers (FSWs) in Urban Zambia Rewa Kohli, Soumyashree Mohanty, Praveen Jadhav, Ramesh Paranjape 2 National AIDS Research Institute, Socio-behavioral Research Department, Pune, India, 2 National AIDS Research Institute, Immunology Department, Pune, India Background: Very little information is available about acceptability of new biological prevention tools such as PrEP and RM among MSM in whom the HIV risk is high. We explored knowledge and willingness about these tools among MSM community in India. Methods: Qualitative study using repeated in-depth interviews among 39 consenting MSM was conducted in Pune. Expert peer and social networking helped recruitment. Interview guide aided discussion. Data collection and analysis occurred concurrently. Results: MSM were between yrs of age, mostly educated till 0 th grade, 2 were highly educated, and 4 MSM were HIV positive, 9 MSM engaged in sex work regularly and 0 occasionally. Few MSM had correct knowledge about HIV. Despite using condoms 23 MSM perceived the risk of HIV infection because of fear of condoms tear/ nonuse owing to liquor consumption. Majority lacked knowledge about other prevention options while 2 mentioned PEP. PrEP was acceptable as it provided added protection in cases of condom nonuse and failure, multiple partners and for a stress free sex life. Majority was fine with daily intake but others wanted it before sex as sex did not happen frequently. Most would continue intake till risk behavior lasts. Concerns about sideeffects and adherence were expressed. Side-effects affecting physical appearance and functioning of internal organs were not acceptable Free/ cost below Rs 500/m was fine. Efficacy should be nearly 00 percent. All MSM were willing to use the RM as it would provide HIV protection. Most MSM were habituated to using lubricants and favored gel formulation. If RM enhances libido it would sell very well. Availability preferred at pharmacy with/without prescription; or NGOs/public hospital at nominal cost/free. Conclusions: It is critical to raise awareness of community about new prevention tools such as PrEP and RM, for successful positioning when efficacious options become available. Tools should be effective, easily available at low cost and be without major side-effects. Jennifer A. Kotlewski, Linda Kimaru, Tyronza Sharkey, Marydale A. Oppert 2, William Kilembe 3, Mubiana Inambao, Hervette Nkwihoreze, Amanda Tichacek 2, Nurelign Ahmed 4, Rachel Parker 2, Susan Allen 2 Rwanda Zambia HIV Research Group, Ndola, Zambia, 2 Rwanda Zambia HIV Research Group, Atlanta, GA, United States, 3 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 4 Rwanda Zambia HIV Research Group, Kigali, Rwanda Background: FSWs in Sub-Saharan Africa are a vulnerable group in the AIDS epidemic. Due to barriers in access to testing and counseling, numerous sexual partners, and little control over safe sex practices with clients, FSWs are at particularly high risk for HIV/STIs. In this study, FSWs in urban Zambia were surveyed and tested for HIV, T. Vaginalis (TV), and syphilis. Methods: FSWs were recruited from known activity hubs through peer outreach in Lusaka and Ndola, Zambia. Interested FSWs received Voluntary HIV Counseling and Testing, RPR serologies for syphilis, and self-administered vaginal swab for TV. A risk assessment survey was administered, including questions on previous testing and use of antiretrovirals (ARV). Results: 733 FSWs were screened over an 8-month period. At baseline, 323 (44%) were HIV positive (HIV+), 39 were HIV negative (HIV-), 9 were indeterminate, and 0 declined testing. 45% had some level of spoken and/or written illiteracy. Among those testing HIV+, 40% reported previous HIV testing and of those 7% were on ARVs. Of HIV+ FSW, 0% were positive for TV and 25% were positive for syphilis. In the HIV- group, 9% were positive for TV and 3% were positive for syphilis. 4% of HIV+ participants reported treatment for STI symptoms in the 2 months before this screening. None of the participants could name health or social services, whether provided by NGO or government, that cater to FSW. Conclusions: Less than half of FSW had been previously tested for HIV. Among HIV+ women who had been previously tested fewer than one in ten were on ART. Low literacy may contribute to poor comprehension of referral services, and fear of stigma may deter FSW from seeking care. Reported treatment for past STI was low, though screening for asymptomatic STI uncovered many cases of syphilis and T.vaginalis. The type of FSW-friendly services provided in this study may be helpful in increasing HIV and STI screening and uptake of ART. Particular attention to low literacy is critical in this effort

204 Posters Posters 3: Key Populations P3. Trends and Factors Associated with Unprotected Anal Intercourse among Young Men who Have Sex with Men in Bangkok, Thailand, Sirirat Lertpruek, Wipas Wimonsate, Sarika Pattanasin, Kesinee Satumay, Wichuda Sukwicha, Jaray Tongtoyai, Kanokpan Pancharoen, Anupong Chitwarakorn 2, Timothy H. Holtz,3 Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Ministry of Public Health, Department of Diseases Control, Nonthaburi, Thailand, 3 Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and Prevention, Atlanta, GA, United States P3.2 Serodiscordant Couples Cohort: An Important Cohort to Explore Correlates of Protection to HIV Infections Sophia Osawe,2, Evaezi Okpokoro 3, Ruth Datiri 2, Grace Choji 2, Felicia Okolo 2, Stephen Umaru 2, Pam Datong 2, Alash le Abimiku 2,3,4 Institute of Human Virology, Jos, Nigeria, 2 Plateau State Human Virology Research Centre, Jos, Nigeria, 3 Institute of Human Virology, Abuja, Nigeria, 4 Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, United States Background: Risk factors for HIV acquisition among young men who have sex with men (YMSM, aged 8-24 years) in Bangkok have been described. We investigated behavior associated with unprotected anal intercourse (UAI) at baseline and during follow-up visits among YMSM enrolled in the Bangkok MSM Cohort Study (BMCS). Methods: Thai men from the Bangkok metropolitan area, aged 8 years who had engaged in sex with another man in the past six months were enrolled and followed up every four months. At enrollment, participants received HIV testing, and were asked about HIV-risk behaviors during the preceding four months using audio computer-assisted self-interviews. We analyzed the factors associated with UAI at baseline and over time using logistic regression and Generalized Estimating Equations (GEE), respectively. Results: We enrolled 72 YMSM during , 62 (87%) contributed at least one-follow-up visit through 203. HIV prevalence was 2%. The overall UAI at baseline was 60% and declined to less than 20% at the 36-month visit, thereafter remaining stable. For each visit increase, we found a 2% decrease in odds for reported UAI regardless of HIV infection. A significant association with UAI at baseline was found among YMSM living with a partner [Adjusted Odd Ratio (AOR) 3., 95% Confidence Interval (CI).7-5.6] compared with YMSM living with family. Factors associated with UAI over time were: 2 sexual partners (AOR 3.6, 95% CI ), current HIV infection (AOR 0.5, 95% CI ), club drug use (AOR.5, 95% CI.-.9), erectile dysfunction drug use (AOR.6, 95% CI.-2.), participation in group sex (AOR.8, 95% CI.5-2.2). Conclusions: YMSM who enrolled in the BMCS exhibited a high level of UAI. The trend over time suggests that intensive HIV-risk reduction counseling and HIV testing campaigns may be associated with a significant reduction in self-reported UAI in this population. However, a longitudinal effect may limit the generalizability. Background: Seronegative partners of a discordant relationship are exposed to HIV and other STIs. Despite availability of ARVs to HIV+ partners, they had varying levels of detectable virus in our cohort and yet their partners remain HIV negative. Studying this more natural model of HIV exposure, which is slightly different from the HESN described by other investigators, can help understand mechanisms of protection to HIV infection. Data from a cohort of discordant couples can explore risk factors in this group and correlates of protection to HIV. Methods: A cohort was developed in October 20 and has been followed up for 2 years and 6 months. Behavioral data collection, medical history, examinations and safety labs were conducted at each of the follow up visits. HIV positive partners were offered CD4 counts and viral loads as well as risk reduction and couples counseling with their partners. Results: Out of 683 screened, 534 were eligible for the study with 257 (48.%) females and 277 (5.9%) males. Condom use was collected at all follow up visits with baseline data showing only 40% of enrollees reporting consistent use of male condoms. Partner viral loads and CD4 counts done revealed that /3 had viral loads above >,000 copies/ml despite being on ARVs and almost 50% had CD4 counts < 350 cells/ µl. Condom use increased slightly during the follow period up to 47%. A total of 22 women (8.6%) became pregnant so far. Recorded HIV and Hepatitis C incidence were.% and 3.7% respectively. Conclusions: Our cohort was made up of couples in a stable home with 99% married. A high number of enrollees expressed their desires to have children explaining the low use of condoms which did not significantly increase through the follow up period despite consistent risk reduction counseling and free condoms. Exposure to HIV is relatively high in the cohort. With documented inconsistent condom use and detectable viral loads this cohorts make an interesting group to look into correlates of protection which is planned. 204 HIV Research for Prevention 204 HIV R4P

205 Posters 3: Key Populations P3.3 Wednesday, 29 October P3.4 Testing for HIV as Individuals or Couples: Preference of MSM in Lagos State Nigeria Adekemi Sekoni, Sikeade Alagbe, Olukemi Odukoya, Kofoworola Odeyemi University of Lagos, Lagos, Nigeria Background: Testing as couples rather than as individuals has been shown to confer added benefits because it allows individuals to also know their partners status and therefore avoid the challenges associated with disclosure of status. Methods: This cross-sectional descriptive study was carried out to assess testing for HIV as individuals and as couples among MSM in eight local government areas of Lagos State, Nigeria using respondent driven sampling to reach 320 men. An interviewer-administered questionnaire was used to collect information after informed written consent was obtained from the participants. Univariate, bivariate and multivariate data analysis was carried out using IBM SPSS 20 software, association was established at p< Results: Majority of the respondents was within the age range years, Christians, self reported bisexual, had post secondary school education and was unemployed. Half of the respondents were single and not in a steady relationship. Majority of the respondents (85.3%) had been tested for HIV, and out of those that have not been tested for HIV, 48.9% are willing to do the test. About half (52.8%) of those who tested as individuals had disclosed their results to their main sexual partners. Less than a quarter of respondents that had been tested did so with their main sexual partner Willingness to test for HIV as a couple (CVCT) among respondents who had previously tested as individuals was poor as only about a third (33.5%) was willing to go for CVCT. Statistically significant association was observed between the age of the respondents, employment status, graded knowledge of HIV/AIDS, knowledge of at least an individual who was tested for HIV or is HIV positive, sexual behavior of respondents and being tested for HIV as individuals but not as couples on bivariate analysis. Conclusions: Couple HIV testing is not popular among our study population, it is necessary to promote this practice among key population in other to achieve zero new infections. Use of Virtual Cohorts in Zambia s Couples Voluntary Counseling and Testing (CVCT) as a Screening Mechanism for Clinical Trials Tyronza Sharkey,2, Rachel Parker,3, William Kilembe,2, Mubiana Inambao,4, Frances Priddy 5, Pat Fast 5, Matt Price 5, Amanda Tichacek,3, Eric Hunter 3, Susan Allen,3 Rwanda Zambia HIV Research Group, Atlanta, GA, United States, 2 Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Emory University, Atlanta, GA, United States, 4 Zambia Emory HIV Research Project, Ndola, Zambia, 5 International AIDS Vaccine Initiative (IAVI)-New York, New York City, NY, United States Background: CVCT, recommended by WHO, is used to identify virtual cohorts (VC) for Zambia Emory HIV Research Project (ZEHRP). VC are at risk groups (HIV concordant negative (M-F-) and discordant couples (DC)) that are followed up and offered various services as standard of care within government and stand-alone clinics. Costs to maintain VC are leveraged by program activities offering these services. Data acquired from these programs are used to calculate seroincidence, follow-up rates and pre-screen potential volunteers for clinical trials. Methods: VC are verbally consented, identified and followed in > 70 centers from Couple HIV status and basic demographics are collected using ZEHRP data collection tools that capture Zambian health indicators, which are reported to the districts. Seroincidence by couple HIV status, % of index partners on/off ARV at seroconversion, and follow-up rates are calculated. Results: Of 48,839 couples tested, 29,8 were identified for VC enrolment (09,677 or 74% M-F- and 7,69 or 2% discordant). 73% of VC were ages 8-40 with 96% cohabiting >3 months. Follow-up rates within VC are 624PY overall with 5083PY in M-F- and 58PY in discordants. In 2 seroconverted couples, 62 were initially discordant and 50 were M-F-. Incidence rates were highest in DC with 5.36/00PY (95% CI ) and lowest in M-F- couples with 0.98/00PY (95% CI ). In DC with linked transmissions (n=48, 77.4%), 27/48 (56%) of index partners had not initiated ARV and 2/48 (44%) were on ARV before seroconversion. Conclusions: VC allow for calculating seroincidence, follow-up rates and is used for pre-screening of volunteers for clinical trials. In discordant couples, VC has illustrated the continued need for inclusion of this high risk group in clinical trials as one size does not fit all. Future plans are to use VC to follow-up other HIV high risk groups in Zambia. VC has the potential to be applied to other diseases/infections for clinical trials and epidemiologic research

206 Posters Posters 3: Key Populations P3.5 Getting a Foot in the Door: Adolescent Retention in a Mock HIV Prevention Clinical Trial P3.6 LB Facilitators and Challenges to ART Adherence among Men who Have Sex with Men (MSM) in Coastal Kenya Elizabeth E. Tolley, Joy N. Baumgartner 2, Sam Field 3, Jennifer Headley, Anna Kaale 4, Anna Minja 4, Sylvia Kaaya 5 FHI 360, Social & Behavioral Health Sciences, Durham, NC, United States, 2 FHI 360, Social & Behavioral Health Sciences, Washington, D.C., DC, United States, 3 FHI 360, Quantitative Sciences, Durham, NC, United States, 4 Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania, United Republic of, 5 Muhimbili University of Health and Allied Sciences, Psychiatry and Mental Health, Dar es Salaam, Tanzania, United Republic of Background: Young women below age 8 are underrepresented in HIV prevention trials, despite being among the most at risk groups in Africa. Their non-inclusion is due in part to ethical concerns. But, adolescents may also face challenges with trial follow-up due to mobility, lower HIV risk perception and less access to sexual and reproductive health (SRH) services. We examine whether retention differs among adolescents aged 5-7 and young women aged 8-2, and what factors might account for this difference. Methods: We enrolled 35 sexually-active, non-pregnant, HIV-negative women, aged 5-2, into a 6-month mock clinical trial in Dar es Salaam, Tanzania, with visits at baseline and months 2, 4, and 6. A mediation analysis assessed direct and indirect effects of age and potential mediators on number of missed visits. Mediators included theoreticallyderived barriers and facilitators to retention. Results: Adolescents held significantly lower risk perception and were less likely to have used SRH services compared to young adults. Both of these factors were positively associated with missed visits, suggesting that adolescents should have missed more visits than adult women. However, controlling for mediators, adolescents were less likely [trending towards significance] to miss visits than adult women. These results suggest that HIV risk perception and previous SRH service use may act as suppressor variables in the relationship between age and retention in a clinical study. Conclusions: Adolescents were no more likely to miss visits than adult women, despite having lower risk perception and less exposure to SRH services. Interventions that realistically increase adolescent risk perception and link them to services - getting their feet in the door - may support their retention in future HIV prevention trials and in SRH services more generally. Also, these analyses suggest that doing so may lead to improvements in retention beyond what is seen in an older cohort of study participants. Murugi Micheni, Andrew Secor 2, Elisabeth van der Elst, Bernadette Kombo, Jane Simoni 2, Don Operario 3, Eduard Sanders, Susan Graham 2 KEMRI-Wellcome Trust Research Programme, Kilifi, Kenya, 2 University of Washington, Seattle, WA, United States, 3 Brown University, Providence, RI, United States Background: In coastal Kenya, 20% of HIV infections occur among MSM, a highly stigmatized group facing many barriers to care engagement and ART adherence. We aimed to identify key facilitators and challenges among HIV-positive MSM, with the goal of developing an intervention to improve clinical outcomes Methods: We conducted individual in-depth interviews (IDI) with purposively sampled HIV-positive MSM and focus group discussions (FGD) with their health providers using semi-structured, open-ended topic guides based on an access-information-motivation-behavioural skills model of adherence, with added focus on provider trust, stigma and discrimination. Detailed interviewer notes and transcriptions were translated into English and reviewed for common factors influencing ART adherence. Results: 30 IDI and 4 FGD were carried out and analysed. Conformity was observed between individual-level factors proposed by the conceptual framework and those reported. Barriers included poverty, limited access to MSM-friendly information and services, poor HIV and ART knowledge, lack of psychosocial support, nondisclosure of HIV status, medication side effects, mental health challenges and substance abuse. Facilitators included economic empowerment, tailored knowledge, peer support, selective disclosure, self-acceptance and pilltaking skills. Resiliency, not identified in the original model, emerged as an important protective factor against stigma and discrimination. A trained ART-experienced peer working with trusted care providers was the favoured means for adherence support. Conclusions: Our final conceptual model comprises factors at the individual, community and policy levels, with access to MSM-friendly services and information, disclosure, self-acceptance and support being key facilitators despite challenging stigma and discrimination. Trust in providers and peers appeared to mediate a hostile social environment and should be fostered through training and structural support. 206 HIV Research for Prevention 204 HIV R4P

207 Posters 3: Key Populations P3.7 LB Sexual Health, Alcohol Use, Childhood Sexual Abuse, and Mental Health Outcomes Among Spanish-speaking Latino MSM in the Northeastern United States. Wednesday, 29 October P3.8 LB Effectiveness of the Community PROMISE and Enhanced Community PROMISE Interventions among Female Sex Workers in the Dakar Region, Senegal Omar Martinez, Elwin Wu 2, Joseph Spadafino 3, Theo Sandfort 2, Andrew Z Shultz, Javier Lopez Rios, Hugo Ovejero 4, Scott D Rhodes 5, Eva Moya 6, Silvia Chavez Baray 6, Brian Dodge 7, Alex Carballo-Dieguez 2 HIV Center for Clinical and Behavioral Studies at Columbia University, Division of Gender, Sexuality, and Health, New York, NY, United States, 2 Columbia University, New York, NY, United States, 3 Arizona State University, Phoenix, AZ, United States, 4 Latino Commission on AIDS, New York, NY, United States, 5 Wake Forest School of Medicine, Winston-Salem, NC, United States, 6 University of Texas, El Paso, TX, United States, 7 Indiana University Bloomington, Bloomington, IN, United States Background: Little is known about the health status of predominantly Spanish-speaking Latino men who have sex with men (MSM). Methods: Between January and March of 204 a cohort of Latino MSM (N=76) was recruited to participate in Latinos en Pareja, an HIV/ STI prevention intervention adaptation study. A multinomial logistic regression model predicting problematic alcohol consumption was carried out; demographic characteristics, sexual risk factors, childhood sexual abuse experiences, and mental health outcomes were included in the model. Results: Prevalence estimates of problematic alcohol consumption in the past 30 days and clinically significant depressive symptoms (CES-D score 0) were 47% and 68%, respectively. Internal consistency reliability coefficients of the CES-D scale were satisfactory (Cronbach α=0.86). Among participants who reported sexual activity before the age of 7 (n=30, 74%), 39 participants (30%) reported childhood sexual abuse. In univariable analyses, characteristics and covariates associated with problematic alcohol consumption included having more than one sexual partner in the past 3 months, engaging in risky sexual behavior (operationalized as condom nonuse in the past 3 months), being in a relationship, reporting intimate partner violence, screening for clinically significant depressive symptoms, and having experienced childhood sexual abuse. In the multinomial logistic regression model, problematic alcohol consumption was predicted by having more than one sexual partner in the past 3 months, engaging in risky sexual behavior, being in a relationship, and reporting intimate partner violence. Conclusions: Further work is needed to develop effective prevention intervention approaches for problematic alcohol consumption among Latino MSM. Given the gap in research on Latino MSM and the high prevalence estimate of childhood sexual abuse among this subpopulation, there is a need to steer effective preventive and treatment interventions to meet the particular needs of this community. Moussa Sarr, Julie Pulerwitz 2, Ibou Thior 2, Marièma Soumaré 3, Ibrahima Traoré 4, Carlos Suarez, Elizabeth King 2, Lindsay Hugues 2, Daouda Gueye 3, Amadou Sougou 3, Aminata Mboup 4, Souleymane Mboup 4 Westat, Rockville, MD, United States, 2 PATH, Washington, DC, United States, 3 AWA, Dakar, Senegal, 4 Cheikh Anta Diop University, Dakar, Senegal Background: In Senegal, the HIV prevalence is % in the general population, but very high among female sex workers (FSWs) with rates up to 30%. The objective of this PATH funded study through a Canadian International Development agency (CIDA) grant is to assess the effectiveness of the Community PROMISE intervention, an effective community-level STD/HIV prevention intervention that relies on role model stories and peers from the target community (Community Promise Only). A structural component was also added to the Community PROMISE activities by targeting bars, bar owners/staff and clients (Enhanced Community Promise). Methods: After adapting and implementing the proposed interventions, we used data from pre and post test surveys of FSWs conducted in the Dakar region to assess differences in consistent condom use with clients and with regular partners. Comparisons were also done between districts of Dakar randomly assigned to Community Promise Only versus Enhanced Community Promise Interventions. Univariate and multivariate logistic regressions were used to assess effectiveness. Results: The average age of the FSW population was years (STD=9.62). Most FSWs were Senegalese (96.7%), and from the Pulaar (3.5%), Sereer (27.4%) and Wolof (2.6%) ethnic groups. Approximately half of FSW (46.9%) never attended school. Overall, we found evidence of increased condom use with clients at post-test versus pre-test (p = 0.04). Consistent condom use increased from 79% at pre-test to 85% at post-test (non-significant) in Community Promise only districts; but reached 93% in Enhanced Community Premise districts (p = 0.006). We did not find evidence of increased condom use with regular partners. Conclusions: Our results showed evidence of increased consistent condom use with clients when both FSWs and clients are targeted by our intervention activities. We see a great potential in diffusing the adapted intervention throughout FSWs communities in Senegal and in other countries in Africa

208 Posters Posters 4: MPT Development P4.0 Drug-drug Interaction Studies Investigating the Impact of Levonorgestrel on Antiviral Potency of Dapivirine Abbey B. Evans, Jonathon Holt 2, Jeremy Nuttall 2, Robin Shattock Imperial College London, Group of Mucosal Infection & Immunity, London, United Kingdom, 2 International Partnership for Microbicides, Silver Spring, MD, United States P4.02 MZC and % TFV Gel: Multipurpose Prevention Approaches Ninochka Jean-Pierre, Patrick Barnable, Larisa Kizima, Aixa Rodríguez, Samantha Seidor, Meredith Clark 2, Gustavo Doncel 2, Melissa Robbiani, Thomas Zydowsky, Natalia Teleshova, José A. Fernández-Romero Population Council, New York, NY, United States, 2 CONRAD Eastern Virginia Medical School, Arlington, VA, United States Background: Intravaginal rings represent an ideal platform to deliver both anti-viral microbicides and hormonal contraception. An intravaginal ring releasing dapivirine (DPV), a potent NNRTI, is currently undergoing late stage clinical efficacy testing. Levonorgestrel (LNG), a synthetic progestogen and hormonal contraceptive, could be used in conjunction with DPV offering the potential of dual protection against HIV infection and pregnancy. Initial drug-drug interaction studies have been performed to determine the potential impact of LNG on the antiviral potency of DPV. Methods: Initially studies were performed to determine the biocompatibility of LNG with common cell lines used to evaluate antiviral potency of candidate drugs. Having determined the maximal compatible dose of LNG in the different cell models, the anti-viral activity of DPV alone or in combination with LNG was determined using TZM-bl, PM- CD4 + T cells and PBMC. Viral replication was assessed by quantitation of luciferase or p24. Results: A concentration of 0µg/ml was selected as the maximal compatible dose for use in all assays. In TZM-bl cells LNG alone showed significant inhibitory activity against HIV- at concentrations >µg/ml. This is likely attributable to the progestogen-driven anti-proliferative effect observed in epithelial cell lines, from which TZM-bl are derived. However LNG had no direct activity against HIV- infection when using PM- CD4 + T cell lines and PBMC, which do not express progesterone receptors. Titration of DPV alone or in the presence of LNG displayed superimposable curves indicating no impact of LNG on the anti-viral activity of DPV. Conclusions: LNG does not alter the anti-viral potency of DPV against HIV- BaL in PM- CD4 + T cells or human PBMC and is therefore unlikely to influence the anti-viral potency of DPV when co-formulated in an intravaginal ring. Background: Next generation microbicides may be broad-spectrum multipurpose prevention technologies with improved efficacy and safety. CAPRISA-004 showed that % TFV vaginal gel prevented HIV- and HSV-2. Here we compare the in vitro and in vivo safety and efficacy profile of % TFV gel versus a microbicide gel (MZC) containing 50 mm MIV-50 (M), 4 mm zinc acetate (Z) and 3% carrageenan (CG). Methods: Increased HSV-2 susceptibility in mice (n=20), TEER in Caco- 2 cells, and macaque vaginal explant histology were used to estimate damage to epithelia. XTT and Cyquant were used to evaluate in vitro cytotoxicity in TZM-bls and PBMCs. Anti-HIV activity was tested against HIV- lab strains (n=5), primary isolates and multidrug resistant strains/ clones (n=28) in TZM-bls or PBMCs. CC 50 and IC 50 values were used to estimate therapeutic indexes (TI=CC 50 /IC 50 ). Anti-SHIV-RT activity (0 4 TCID 50 /explant) of diluted gels was assessed in macaque vaginal explants. In vivo anti-hsv-2 activity (5x0 3 pfu/mouse, n=20/treatment) was evaluated in mice (gel applied h before and after HSV-2 vaginal challenge). Levels of TFV and TFV-DP in mouse cervicovaginal tissues were quantified by LC-MS/MS. Fisher s exact test (P< 0.05) was used for statistical comparison in all HSV-2 murine models. Kruskal-Wallis and Dunn s Multiple Comparison test (P< 0.05) in the explant model. Results: Both gels were safe. However, diluted % TFV reduced TEER values in Caco-2 monolayers. MZC showed greater in vitro antiviral activity (2 to 80-fold) than % TFV gel against HIV- in TZM-bls. Both gels showed good TI (>25-800) in PBMCs, except for one and two MDR HIVs with %TFV and MZC, respectively. MZC fully protected explants from SHIV-RT (p< 0.009) with greater anti-shiv-rt activity than % TFV gel. MZC fully protected mice from HSV-2 (p< 0.000), but the % TFV gel did not. The lack of protection could be associated with subtherapeutic levels of TFV and TFV-DP in mouse tissues. Conclusions: MZC is a promising microbicide candidate for clinical use. 208 HIV Research for Prevention 204 HIV R4P

209 Posters 4: MPT Development P4.03 Wednesday, 29 October P4.04 Formulation Development of a Combination Silicone Elastomer Ring for Vaginal Delivery of Dapivirine and Levonorgestrel Susan Fetherston,2, Clare McCoy 2, Diarmaid Murphy 2, Peter Boyd 2, Andrew Brimer 3, Jonathon Holt 3, Brid Devlin 3, Wendy Blanda 3, Jeremy Nuttall 3, Chris Gimour 3, Karl Malcolm 2 University of Ulster, School of Pharmacy and Pharmaceutical Sciences, Coleraine, United Kingdom, 2 Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 3 International Partnership for Microbicides, Silver Spring, MD, United States Matrix Vaginal Ring Formulations that Maintain Target in-vitro Release Rates of Dapivirine and Levonorgestrel (Alone or in Combination) over 90 Days Jonathon Holt, Andrew Brimer, Susan Fetherston 2, Peter Boyd 3, Brid Devlin, Karl Malcolm 3 IPM, Silver Spring, MD, United States, 2 University of Ulster, Coleraine, United Kingdom, 3 Queen s University Belfast, Belfast, United Kingdom Background: IPM is currently evaluating a 25mg dapivirine (DPV) vaginal ring (VR) in Phase III clinical trials as a 30-day HIV microbicide product. A second generation VR delivering both DPV and levonorgestrel (LNG) is being developed as a multi-purpose prevention technology (MPT) offering simultaneous HIV prevention and hormonal contraception for at least 60 days. In order to maintain DPV release rates for 60 days at levels likely to provide protection, it will be necessary to increase the DPV loading beyond 25mg. Methods: Eleven prototype, silicone elastomer (Nusil MED-4870), matrix VRs containing various loadings of DPV (0, 00, 50 and 200mg) and/or LNG (0, 6 and 32mg) were manufactured by injection molding at 60 C for 60 sec. In vitro release testing under sink conditions was performed for at least 60 days. Release of DPV and LNG from the VRs was quantified by HPLC-UV. Results: In vitro release of DPV and LNG obeyed root time kinetics for all VRs tested, as expected for matrix-type devices and indicative of a permeation-controlled release mechanism. Increasing the DPV or LNG loading significantly increased the release rate for the same active (P < 0.00). The addition of LNG (6 or 32 mg) in the VRs did not significantly influence DPV release. LNG release was significantly increased by the presence of DPV, as compared to LNG only rings with equivalent LNG loading. Each formulation tested exceeded the in-vitro release target for DPV (200 µg at d60). Day 60 DPV release was µg, µg, and µg for the 00, 50 and 200 mg DPV VRs respectively. Conclusions: The data confirm the potential for development of a MPT VR strategy based on a matrix ring containing DPV and LNG. Dapivirine load of 200 mg should maintain requisite levels of dapivirine for 60 days in-vitro. In-vitro release of LNG is enhanced by the presence of DPV. In-vivo testing of DPV-LNG rings will be necessary in order to test drugdrug release effects under physiological conditions Background: IPM s silicone-based Dapivirine Vaginal Ring is in Phase III trials as a vaginal microbicide for month continuous use. New formulations have been assessed with in-vitro drug release of greater than 90 days. By extending the use duration per ring, the overall production cost per patient per month may be decreased to nearly one third that of the monthly dapivirine vaginal ring. Methods: Silicone vaginal rings manufactured at Queen s University Belfast using similar manufacturing processes as IPM s monthly dapivirine vaginal ring were produced as either dapivirine (DPV) only (25 and 200mg) or a combination of DPV (200mg) & LNG (32 mg). Nusil Med-4870 (a Pt-catalyzed, addition cured silicone polymer) was used to produce rings with an overall diameter of 5.67cm, cross-section diameter of 7.82mm and weighing 8g (60 C cure for 60 sec). Daily in-vitro release (IVR) of DVP and/or LNG was assessed up to 92 days in 50mL of : IPA: water. Media was assayed by HPLC for DPV and LNG. Results: The DPV IVR target was 05 µg, which is the day 28 (25 mg) Dapivirine Vaginal Ring release rate. Formulations containing 200 mg DPV exceeded the target (d30 = 628 µg; d60 = 437 µg; d92 = 30 µg). In addition, when formulated in combination with LNG, DPV IVR (µg/ day) remained above the target (d30 = 634; d60 = 454; d92 = 299). LNG IVR was also above target (35 µg/day) for all 92 days tested (d30 = 55; d60 = 9; d92 = 46). The day release for a 200mg DPV ring (432 µg DPV alone; 6038 µg DPV with LNG) and for a 32 mg LNG ring (684 µg) remained at levels below those established as safe in nonclinical toxicity studies. Conclusions: The IVR data presented indicate that it may be possible to produce a DPV-only microbicide ring and a DPV -LNG combination microbicide and contraceptive ring for use over 90 days using a silicone matrix ring. Furthermore, within the given range, elevation of DPV loading (or co-loading with LNG) does not increase in-vitro release to unsafe levels

210 Posters Posters 4: MPT Development P4.05 In Vitro Design and Drug Delivery of Combination Dapivirine and Contraceptive Hormone Ethylene Vinyl Acetate (EVA) Vaginal Ring Prototypes Jonathon Holt, Tiffany Derrick, Andrew Brimer, Andrew Loxley 2, Brett Breaker 2, Matthew Bigert 2, Michael Grieco 2, Bruce Frank 2, Brid Devlin IPM, Silver Spring, MD, United States, 2 Particle Sciences Inc., Bethlehem, PA, United States P4.06 Evaluation of the Factors Contributing to Levonorgestrel Binding in Addition Cure Silicone Elastomer Vaginal Rings Karl Malcolm, Diarmaid Murphy, Clare McCoy, Peter Boyd, Susan Fetherston 2, Andrew Brimer 3, Jonathon Holt 3, Wendy Blanda 3, Brid Devlin 3, Jeremy Nuttall 3, Chris Gimour 3, Tiffany Derrick 3 Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 University of Ulster, School of Pharmacy and Pharmaceutical Sciences, Coleraine, United Kingdom, 3 International Partnership for Microbicides, Silver Spring, MD, United States Background: Dapivirine (DPV) is currently formulated in a silicone ring for Phase III clinical trials as a vaginal microbicide. NuvaRing (a marketed vaginal contraceptive ring) is EVA-based and has many of the characteristics desired for a DPV/hormone combination ring. This work describes the design and in vitro testing of EVA vaginal ring prototypes containing DPV and various contraceptive hormones. Methods: Two contraceptive hormone options were co-delivered with DPV from either matrix or core-sheath designs: Etonogestrel (ETO) combined with Ethinyl Estradiol (EE); Levonorgestrel (LNG). In vitro elution (IVE) targets were based on the DPV ring currently in Phase III trials, NuvaRing (ETO and EE) and literature (LNG). The 60d IVE profile was determined for matrix ring designs for all hormones and for coresheath rings with various loads of LNG and DPV in the core, and several sheath thicknesses (blank or with 0mg of DPV). Results: Matrix ring prototypes released DPV and all hormones with first order kinetics, however, the day bursts for ETO and LNG were high (>000 µg). Rings loaded with 25 mg DPV released drug at target rates on day 60 and the day burst was within expected limits. Core sheath designs had a blunted initial release compared to the matrix rings but the anticipated zero order kinetics was not achieved with a loaded core and blank sheath. DPV loaded sheaths further increased the overall DPV release across 60 days but only the lower LNG target (35 ug/day) was achieved with any configuration of LNG load. Stability tests indicated a need for refrigerated storage conditions to prevent migration of LNG to the ring surface and transfer to the packaging. Conclusions: These studies demonstrate that EVA would be a suitable polymer for the dual delivery of dapivirine and contraceptive hormones from a vaginal ring under refrigerated conditions. These conditions were deemed unsuitable for use in developing countries due to cost and therefore alternative polymers are being evaluated. Background: Following progress of the dapivirine (DPV)-releasing silicone elastomer (SE) vaginal ring (VR) into Phase III clinical studies, there is now interest in developing next-generation rings that additionally provide contraception. Levonorgestrel (LNG) is a safe and effective progestin that is being widely considered for use as a hormonal contraceptive agent in future multipurpose prevention technology (MPT) products. Although LNG has previously been incorporated into various controlled release SE devices, minimal attention has focused on its propensity to irreversibly react with addition cure SE systems. Here, for the first time, we investigate this LNG binding phenomenon and outline strategies for overcoming it. Methods: VRs containing various loadings of DPV and LNG were manufactured and in vitro release assessed. Different LNG-only SE samples were also prepared to assess the following parameters: (i) addition cure vs. condensation cure SEs; (ii) different types of addition cure SEs; (iii) mixing time, (iv) cure temperature, (v) cure time; and (vi) LNG particle size. After manufacture, the LNG-only samples were assayed for total drug content using a solvent extraction method. The SE curing reaction and the LNG binding reaction was probed using nuclear magnetic resonance (NMR) spectroscopy. Results: Under certain drug/formulation/processing conditions, LNG was not recoverable from VRs. Further studies using non-ring samples showed that: (a) the phenomenon was only observed with addition cure SEs (and not condensation cure SEs); (b) the extent of binding was dependent upon the type of addition cure SE; (c) micronised LNG showed significantly greater binding than nonmicronised LNG; (d) the extent of binding correlated with increased mixing time, cure time and cure temperature. Conclusions: Careful control of the API characteristics, the SE composition, and the manufacturing conditions will be necessary to establish a practical VR formulation for controlled release of LNG. 20 HIV Research for Prevention 204 HIV R4P

211 Posters 4: MPT Development P4.07 Thermal Properties and Eutectic Behaviour of Dapivirine in Combination with Steroid Hormones and Other Antiretrovirals Wednesday, 29 October P4.08 Selection of a Hormonal Contraceptive Agent for Inclusion with Dapivirine in a Silicone Elastomer Vaginal Ring Karl Malcolm, Clare McCoy, Diarmaid Murphy, Peter Boyd, Andrew Brimer 2, Jonathon Holt 2, Brid Devlin 2, Wendy Blanda, Jeremy Nuttall 2, Chris Gimour 2 Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 International Partnership for Microbicides, Silver Spring, MD, United States Clare McCoy, Peter Boyd, Susan Fetherston 2, Ian Major, Diarmaid Murphy, Brid Devlin 3, Andrew Brimer 3, Jonathon Holt 3, Jeremy Nuttall 3, Chris Gimour 3, Wendy Blanda 3, Karl Malcolm Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 University of Ulster, School of Pharmacy and Pharmaceutical Sciences, Coleraine, United Kingdom, 3 International Partnership for Microbicides, Silver Spring, MD, United States Background: Combination drug products can display thermal behaviour that is more complex than for the corresponding single drug products. For example, the contraceptive vaginal ring (VR) Nuvaring contains a eutectic (lowest melting) composition of etonogestrel (ETN) and ethinyl estradiol. Here we report the predisposition of dapivirine (DPV) to form reduced melting/eutectic mixtures when combined with other contraceptive hormones and antiretrovirals, and discuss the implications for development of combination microbicide and multipurpose prevention technology (MPT) products. Methods: Binary mixtures of DPV with darunavir (DRV), levonorgestrel (LNG), ETN or maraviroc (MVC) were prepared either by physical mixing or by solvent evaporation. Selected binary mixtures were also incorporated into silicone elastomer (SE) VR devices. Thermal behavior of the mixtures was analyzed using differential scanning calorimetry (DSC) operating in standard heating ramp mode (0 C/min). DSC data were used to construct two component phase diagrams for each binary system. Results: Drug mixtures typically showed reduced melting transitions for both drug components, with clear evidence for a eutectic mixture at a well-defined intermediate composition. Eutectic temperatures and compositions for the various mixtures were: 40% DPV / 60% ETN - 70 C; 25% DPV / 75% MVC - 72 C; 65% DPV / 35% LNG - 92 C. In each case, the eutectic composition was also detected when the drug mixtures were incorporated into SE VRs. For the DPV/DRV system, the thermal behaviour is complicated by desolvation from the darunavir ethanolate polymorph. Conclusions: When DPV is combined with small molecular weight hydrophobic drugs, the melting temperature for both drugs is typically reduced to a degree dependent on the composition of the mixture. At specified compositions, a low melting eutectic system results. The formation of eutectic behavior in binary drug systems needs to be carefully characterised in order to define product performance and drug release. Background: A vaginal ring (VR) that can prevent both unintended pregnancy and the spread of sexually transmitted infections has the potential to significantly improve women s sexual and reproductive health. Here, we report in vitro evaluation of four hormonal contraceptive compounds - levonorgestrel (LNG), ethinylestradiol (EE), etonogestrel (ET) and nestorone (NES) - for co-formulation with dapivirine (DPV) in a silicone elastomer (SE) vaginal ring device. Methods: The influence of each steroid hormone on the SE cure characteristics was assessed using flow rheology. The thermal stability of each API and the potential for drug-polymer and drug-drug interactions were assessed using thermal gravimetric analysis (TGA) and differential scanning calorimetry (DSC). Matrix-type and reservoir-type VRs containing various loadings of each steroid were manufactured using both a condensation-cure and an addition-cure SE systems and in vitro release rates assessed. Results: TGA confirmed all steroids were stable in the temperature range required for DPV ring manufacture. DSC analysis confirmed that all steroids showed eutectic behaviour when combined with DPV. Rheological analysis demonstrated that EE and ET, but not LNG or NES, inhibited curing of addition-type SE system. All steroids could be effectively released from the SE VRs. However, only LNG release could be sustained for the target 30 day period. Conclusions: Based on the results of these studies, LNG was selected as the most suitable contraceptive agent for further development with DPV in a dual-purpose vaginal ring. 2

212 Posters Posters 4: MPT Development P4.09 P4.0 The Need for Multi-product Technologies for Women Participating in Vaginal Microbicide Trials Duduzile E. Ndwandwe, Nathlee S. Abbai, Handan Wand 2, Gita Ramjee South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 University of New South Wales, The Kirby Institute, Sydney, Australia Background: Sub-Saharan Africa continues to bear a towering share of the global HIV burden with 70% new HIV infections. HIV prevalence among young women remains more than twice as high as among young men throughout sub-saharan Africa Therefore, it is important to perform a needs assessment for women enrolled in HIV prevention trials as a strategy to increase adherence to investigational products. The aim of this study is to assess commitment to prevention interventions by measuring pregnancy and HIV incidence in relation to product adherence. Methods: This study included data collected from women (n=456) enrolled in the Carraguard vaginal microbicide gel trial. Associations between gel adherence and pregnancy and HIV incidence in women during the study follow-up were described using Kaplan-Meier and Cox regression analysis. Statistical analysis was performed using STATA release 2.0 (College Station, Texas, TX, USA). Results: The highest crude incidence rates for pregnancy (6.7 per 00 person year (py), 95% Confidence interval (CI) 5., 8.9) and HIV (8.5 per 00/py, 95% CI: 6.2,.6) were reported for women that were in the 2 nd quartile of % gel used with condoms (35-< 55%). After adjusting for age, number of sexual partners in the last 3 months, number of sex acts in the last week and marital status; women that had very low gel adherence with condom use were at increased risk for incident pregnancy ( st quartile < 35%, Hazard Ratio (HR) 2.4, 95% CI: and HIV infections (2 nd quartile 35-< 55%, HR: 3.5, 95% CI:.8, 7.0) when compared to high adherers (>75% gel adherence with condom use, HR: per 00 p/y respectively). Conclusions: These findings suggest that adherence to study product and condom promotion is critical to prevent infection with HIV as well as reduce pregnancy incidence. Multipurpose Prevention of HIV and Pregnancy: Perspectives of Users, Providers and Community Advisors in Zimbabwe and Malawi Cynthia Woodsong, Petina Musara 2, Adlight Chandipwisa 2, Elizabeth Montgomery 3, Patty Alleman 4, Zvavahera Mike Chirenje 2, Tsungai Chipato 2, Francis Martinson 5, Irving Hoffman 6 International Partnership for Microbicides, Silver Spring, MD, United States, 2 UZ-UCSF Collaborative Research Programme, Harare, Zimbabwe, 3 RTI International, Women s Global Health Imperative, San Francisco, CA, United States, 4 USAID, Office of Population and Reproductive Health, Washington, DC, United States, 5 UNC Project, Lilongwe, Malawi, 6 UNC, UNC Hospitals, Chapel Hill, NC, United States Background: Prevention of HIV and pregnancy could be accomplished with a single Multipurpose Prevention Technology (MPT), and a number of topical vaginal products are currently under development. There is limited information on the interests of potential MPT users and those advising on use, particularly in sub-saharan Africa. Rather, assumptions about interest in such a product are inferred from what is known about acceptability and use of vaginal microbicides or contraceptives. Methods: This paper presents data on concerns and preferences for multipurpose prevention of HIV and pregnancy. Data were collected in two on-demand microbicide gel studies in Malawi and Zimbabwe. Participants were women using candidate vaginal products, their male partners, health professionals and community stakeholders. All participants provided an interview which was audio-recorded, transcribed, coded for content, and analyzed for key themes concerning interest and concerns about multipurpose prevention. Results: Participants indicated strong interest in a vaginal HIV prevention product that could also prevent pregnancy. Key reported advantages included convenience, the potential to avoid side effects experienced with current contraceptive methods, concerns about long term effects of contraceptives, and concerns about the health burdens of HIV infection during pregnancy. The main disadvantage of an MPT was recognition that while interest in preventing HIV is constant, contraceptive needs change over time. Conclusions: The study provides much-needed primary data on interest in an MPT to prevent HIV and pregnancy. This interest may be further strengthened if a product is also available for prevention of only HIV. Health professionals and community advisors may be more likely to recommend an MPT, and women and men may be more willing to use an MPT, if they can be reassured that use will have no long-term effect on fertility, and they can switch from single to multipurpose prevention as needs change over time. 22 HIV Research for Prevention 204 HIV R4P

213 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.0 ABSTRACT WITHDRAWN P5.02 Acceptability of a Polyurethane Tenofovir Disoproxil Fumarate (TDF) Intravaginal Ring for HIV Prevention Dana L. Watnick, Marla J. Keller, Lilia Espinoza, Betsy C. Herold, Laurie J. Bauman Albert Einstein College of Medicine, Bronx, NY, United States Background: There is urgent need for female-controlled HIV prevention strategies such as topical pre-exposure prophylaxis. Unfortunately, intravaginal ring (IVR) development has had little input from users. As part of an ongoing Phase TDF IVR safety and pharmacokinetic trial among 30 US women, we are qualitatively investigating how HIV risk perception and ring acceptability, specifically related to partner concerns, menstruation and timing of ring exchange, may affect user receptivity to ring use. Methods: Female participants (8-45 years) are randomized to a polyurethane TDF or placebo IVR for 4 days of continuous use and participate in two in-depth interviews. Data are collected and analyzed using a Grounded Theory approach. We report results for the first 0 women who completed the trial (6 TDF, 4 placebo). Results: No one perceived herself at risk for HIV, but all conceded infidelity ( even spouses might cheat ) and 3 identified rape, sex work and living in Africa as HIV risk factors. Trusting one s partner to always be faithful and absence of a sexual relationship were reasons to forego ring use. Issues with partner acceptability were male drug exposure, interference with sex, suspicion of her cheating, and facilitating his cheating. Most women (n=8) had concerns about ring use during menses: potential infection from blood on the ring; the body s need to clean out toxins (including the drug itself); tampons soaking up medication; blood blocking the outward flow of drug; and not needing a ring because of abstinence during menses. Most (n=8) preferred to remove the ring monthly before menses or to clean it to avoid infection or funky odor. Seven subjects questioned ring effectiveness the longer it was left inside the vagina. Conclusions: Lack of perceived HIV risk and partner concerns are potential barriers to ring uptake. Menstruation and frequency of ring exchange are inextricably linked around the monthly cycle; half prefer monthly ring changes. 23

214 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.03 Jatropha sp. Extracts Induces CD4 Internalization and Inhibits HIV- Entry Paola Silveira, Rodrigo Orlandini 2, Rodrigo Cunha, Thais Barbizan 3, Luis Pianowski 3, Luis Da Silva 2, Amilcar Tanuri, Renato Aguiar Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, 2 Federal University of São Paulo, Ribeirao Preto, Brazil, 3 Kyolab Laboratories, Valinhos, Brazil P5.04 Development of a Novel Bioadhesive Microbicide Gel Formulation for Prophylactic Protection against HIV and HSV-2 Gita N. Shankar, Gaurav Bhatia, Carsten Alt 2 SRI International, Pharmaceutical Sciences, Menlo Park, CA, United States, 2 Palo Alto Institute for Research and Education, Palo Alto, CA, United States Background: In 202, were recorded approximately 2.3 million of new HIV infections. These numbers show the need to develop drugs that can prevent the infection, since an effective vaccine for HIV is not available. Microbicides are an alternative for primary prevention and control the HIV epidemic. Here, we screened extracts from the Brazilian plant Jatropha sp. to evaluate cytotoxicity and potential antiviral activity against HIV- in TCD4 cells. Methods: The cytoxicity and antiviral activity of Jatropha sp. extracts were initially screened in TCD4+ lymphocytes. Cells were pre-exposed to extracts and the HIV infectivity was measured by luciferase reporter gene cloned in the HIV genome.the downmodulation of CD4 receptor was checked by flow cytometry. To elucidate the molecular mechanisms of CD4 internalization several mutants of CD4 tail were transfected in HeLa cells treated and the CD4 staining. CD4 degradation was evaluated by the lysosome inhibitor and immunoblotting. Cellular morphology after treatment was checked by electron microscopy and the cytokines produced by treated cells was measured by Bioplex system. The cell activation was evaluated by flow cytometry with specific antibodies. Results: The THS fraction decreased HIV- infectivity up to 80% in a dose-dependent manner with no cytoxicity. THS treatment induces internalization of CD4 receptor in PBMC cells, direction to early endosomes, followed by lysosomal degradation. The CD4 internalization promoted for THS treatment was mediated by PKC activation and consequent phosphorylation of CD4 tail serine residues. Our results of electron microscopy showed that the THS stimulate macropinocytic. Finally, it was observed that the compound activated cells with high expression of CD69 and increased secretion of IL-8 and MIP-beta. These data suggests that THS treatment does not causes T cell anergy. Conclusions: THS induces CD4 internalization and inhibits HIV entry suggesting that compounds can be potentially used as microbicide to prevent HIV transmission. Background: Over-the-counter access to an inexpensive topical microbicide could reduce transmission of human immunodeficiency virus (HIV) and would increase women s control over their health and eliminate the need to obtain their partners consent for prophylaxis. Chronic infection with herpes simplex virus-2 (HSV-2) has been shown to facilitate HIV infection and speed the progression to disease. Our objective is to develop a drug formulation that adheres to the vaginal surface for extended time, while protecting against both HIV and HSV-2. We present here development studies of a novel bioadhesive vaginal delivery platform SR-2P which is composed of two FDA-approved polymers, acting as a depot for two approved antiviral drugs, acyclovir and tenofovir. Methods: Prototype gels were developed and evaluated for ph, osmolality, buffering capacity and viscosity under simulated vaginal semen dilutions, and bioadhesivity using minipig vaginal tissues. Release profile of drug loaded SR-2P was performed through porcine vaginal mucosa. Vaginal irritation studies were performed in mice and rabbits. Results: Results show that SR-2P retains its structure and bioadhesivity upon dilution in presence of simulated fluids under stress conditions. The in-vitro release profile of SR-2P demonstrated that ~40% tenofovir and ~38% acyclovir was retained in the porcine vaginal mucosa, in the presence of vaginal semen fluids even after 6 hours of contact time. SR-2P caused minimal irritation in a mouse and a rabbit vaginal model. Our results indicate that SR-2P containing acyclovir and tenofovir protects Vero cells from infection with HSV-2 in vitro and mice from HSV-2 in vivo. Conclusions: This preliminary study demonstrates that SR-2P gel could be used as a vaginal microbicide for prophylaxis against vaginal HIV and HSV-2. Research reported in this publication was supported by NIAID/ NIH Award No.AI The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. 24 HIV Research for Prevention 204 HIV R4P

215 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.05 Design of Biodegradable Nanoparticles for Vaginal Delivery of the anti-hiv Protein Griffithsin Haitao Yang,2, Jing Li, Charlene S. Dezzutti 2, Kenneth Palmer 3, Lisa C. Rohan,2,4 University of Pittsburgh School of Pharmacy, Department of Pharmaceutical Sciences, Pittsburgh, PA, United States, 2 Magee Women Research Institute, Pittsburgh, PA, United States, 3 University of Louisville School of Medicine, Louisville, KY, United States, 4 University of Pittsburgh School of Medicine, Department of Obstetrics, Gynecology, and Reproductive Sciences, Pittsburgh, PA, United States P5.06 Nanodelivery of CSIC for Enhanced Solubility and Rapid Macrophage Uptake Tiantian Gong,2, Michael Parniak 3, Phalguni Gupta 4, Lisa Rohan,2,5 Magee Womens Research Institute, Pittsburgh, PA, United States, 2 University of Pittsburgh School of Pharmacy, Pittsburgh, PA, United States, 3 University of Pittsburgh, Department of Microbiology & Molecular Genetics, School of Medicine, Pittsburgh, PA, United States, 4 University of Pittsburgh Graduate School of Public Health, Department of Infectious Diseases and Microbiology, Pittsburgh, PA, United States, 5 University of Pittsburgh, Department of Obstetrics, Gynecology and Reproductive Sciences, Pittsburgh, PA, United States Background: Griffithsin (GRFT) is an anti-hiv protein which blocks gp20 binding to CD4 receptor-expressing cells and binds to mannoserich glycans on viral glycoproteins (gp20, gp4, and gp60). Its broad-spectrum anti-hiv activity and unique safety profile support its development as a topical microbicide. However, its proteinous nature and poor tissue permeability limit its antiviral activity to the vaginal lumen. That leaves HIV targeted cells within submucosa and deeper vaginal tissue vulnerable without the protection of GRFT, which will consequently diminish its potential as a topical microbicide product. The objective of this study was to use poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) to enhance the penetration of GRFT leading to more efficient delivery of this anti-hiv drug. Methods: A double emulsion-solvent evaporation method was utilized to manufacture GRFT loaded PLGA NPs. Particle size, size distribution and zeta potential of NPs were determined utilizing dynamic light scattering. Images of the nanoparticles were taken by transmission electron microscopy (TEM). The amount of GRFT encapsulated in the nanoparticles was determined by HPLC method. The anti-hiv activity and toxicity of GRFT nanoparticles was studied in a TZM-bl cell based assay. Results: The developed GRFT loaded PLGA NPs showed nearly spherical shape with a particle size of nm and narrow size distribution (PDI < 0.2). Particle size was confirmed using TEM. Particle zeta potential was -30 mv. Encapsulation efficiency for GRFT was found to be 42%. In vitro studies showed that the IC 50 of GRFT loaded NPs was comparable with GRFT (»3 nm). In these same studies cells viability of both PLGA NPs and GRFT loaded NPs was completely maintained. Conclusions: These studies represent the incorporation of GRFT into a PLGA NPs system. The encapsulation of GRFT into PLGA NPs did not impact its anti-hiv activity and safety. NP incorporation of GRFT may enhance its tissue penetration leading to increased protection. Background: CSIC is a hydrophobic, tight-binding, non-nucleotide reverse transcriptase inhibitor with high potency against HIV- in vitro. To enhance its aqueous solubility a three phase nanoparticle engineering technology was used to manufacture CSIC nanocrystals (NC) with acceptable properties for intravaginal application. Methods: Nanocrystals were prepared using co-precipitation, hydration, and stabilization steps. Particle size distribution, polydispersity index (PdI), and zeta potential were determined using dynamic light scattering. NC drug release and stability were assessed. Surface morphology was investigated by transmission electron microscopy (TEM). Differential scanning calorimetry (DSC) was used to monitor crystal properties. Drug content was analyzed by HPLC-UV. Biological attributes including mucin interaction, bioactivity, and macrophage uptake were studied. Results: Nanocrystal stabilization was achieved through use of a combination of Pluronic F98 (0.3mg/mL) and hydroxypropyl methylcellulose (HPMC) E5 (5mg/ml). NC obtained using this system had a particle size of 260nm (PdI < 0.4) and surface charge of -7.2mV. Particle size was maintained for 8h in a liquid state, and for at least 2 month after lyophilization. An amorphous state following co-precipitation was confirmed using DSC. Following hydration and stabilization, needleshaped crystals resembling the raw CSIC powder were obtained (confirmed by DSC and TEM). Aqueous solubility of the NC CSIC form was increased by ~500 fold. Encapsulation efficiency was >80% (0.7mg/mL) and 75% of loaded CSIC was released from NC within 4 days. No change in particle size was observed in the presence of mucin indicating limited mucoadhesive properties. NC formation resulted in no loss of bioactivity. In vitro studies showed rapid macrophage uptake via an energy-dependent pathway. Conclusions: Nanocrystals can be used to improve CSIC aqueous solubility and drug targeting which are critical for drug localization at the target site. 25

216 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.07 Vaginal and Rectal Use of OTC Lubricants: Safety in the Macaque Model P5.08 NMR Metabonomics in an in vitro Model of HIV- Latency Dorothy L. Patton, Yvonne Sweeney, Sharon Hillier 2 University of Washington, Obstetrics & Gynecology, Seattle, WA, United States, 2 Magee Women s Research Institute, Pittsburgh, PA, United States Thato P. Nonodi, Debra Meyer University of Pretoria, Biochemistry, Pretoria, South Africa Background: Over-the-counter (OTC) lubricants are marketed for use during vaginal and anal receptive intercourse but these products have not been extensively studied for safety. The macaque model provides a standardized means for evaluation of vaginal and rectal product safety. Methods: Four OTC lubricants available in the USA were evaluated including three aqueous-based personal lubricants (hypo-osmolar FemGlide, iso-osmolar Pre-Seed, hyper-osmolar KY Jelly), and hyperosmolar, lipid based Elbow Grease cream. Six macaques completed each study arm. Animals underwent 4 daily applications of each lubricant, with measurements collected prior to and 30-minutes after each use, and 24-hours after the final exposure. The integrity of the mucosal epithelium at the site of product use was assessed by colposcopy following vaginal use and assessment of epithelial sloughing in rectal lavage samples following rectal application. The microflora and ph was also assessed before and after daily exposure to test agents. Results: No evidence of product-related epithelial disruption was noted with vaginal use of any of the four products. Two products were associated with increased epithelial sloughing in rectal lavage samples after use (KY Jelly and Pre-Seed). The vaginal and rectal microbiota was not grossly affected by product use; however repeated vaginal use of KY Jelly suppressed the presence of beneficial lactobacilli in half of the animals. ph measures were only slightly and temporarily influenced by the product ph in all cases. Conclusions: OTC personal lubricants had variable effects on the genital and rectal tracts of pigtailed macaques. While FemGlide and Elbow Grease Cream had acceptable safety profiles with vaginal and rectal use, some other OTC lubricants adversely affected the vaginal microbiota or were associated with epithelial sloughing following rectal application. Given the wide use of personal lubricants during sexual activity, additional safety studies are warranted. Background: HIV latency is viewed as one of the important reasons why eradication of infection has been elusive thus far. Studies on how the virus affects immune system cells during its active and latent phases could provide targets for drug development as well as a means to monitor the progress and status of infection. Since the implementation of omics studies, whole cells/systems/pathways have been elucidated by the use of high throughput analytical tools like nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS). Methods: In this study metabonomics analysis of U cells (a promonocytic HIV- latency and chronic infection model) was used to investigate and compare HIV- latency to active infection. The uninfected parent cell line (U937) served as a necessary control. Metabolites from these cells were extracted and analysed by NMR. Results: Six important metabolites were detected; these metabolites were identified using relevant databases and found to be lactate, lipid, diethylthiophosphate, methylsuccinate, choline and methylacetoacetic acid. Changes in the peak height of lactate peak between the different viral phases presents as a possible marker of infection status. Conclusions: Lactate, is a product of glucose metabolism which is already known to be influenced by active HIV- infection. Data presented here suggests that even in latency the virus appears to have an effect on glucose metabolism. 26 HIV Research for Prevention 204 HIV R4P

217 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.09 Drug-drug Interactions between the Dapivirine Vaginal Ring (Ring-004) and Miconazole Nitrate Vaginal Capsule (Gyno- Daktarin ) P5.0 Broad-spectrum Anti-HIV- Activity of Anionic Carbosilane Dendrimers and Synergy in Combination with Maraviroc and Tenofovir as Topical Microbicide Annalene Nel, Wouter Haazen 2, Marisa Russell, Jeremy Nuttall 3, Neliette Van Niekerk, Nicoline Treijtel 4 International Partnership for Microbicides, Clinical Affairs, Paarl, South Africa, 2 SGS Life Science Services, Clinical Pharmacology Unit Antwerpen, Antwerp, Belgium, 3 International Partnership for Microbicides, Product Development, Silver Spring, MD, United States, 4 Kinesis Pharma BV, Clinical Pharmacokinetics, Breda, Netherlands Background: The Dapivirine Vaginal Ring-004 (25 mg dapivirine) is a topical microbicide currently being evaluated in Phase III trials. Given the potential for interactions with co-administered vaginal products, pharmacokinetic assessments were performed during co-use of Ring- 004 and the antifungal miconazole nitrate. Methods: An open-label, randomised, crossover trial was conducted among 36 healthy, HIV-negative women, aged 8-40 years. Participants used dapivirine Ring-004 for 28 days, alone or together with a single dose of miconazole nitrate (200 mg vaginal capsule). A single dose of miconazole nitrate was administered alone in a third treatment period. Washout periods of 3 weeks were included between treatments. Dapivirine and miconazole concentrations were determined in plasma and vaginal fluid (CVF) samples, and residual dapivirine levels were assessed in used rings. Results: A single vaginal dose of miconazole nitrate at the time of dapivirine ring insertion increased the systemic exposure (C max and AUC) of dapivirine by approximately 20%, but decreased dapivirine CVF levels up to 4 days post-ring insertion (decreases of 26% in C max and 69% in AUC 0-24h ). No significant difference was observed between dapivirine ring residual levels with and without concomitant miconazole, suggesting similar dapivirine release. Dapivirine CVF levels remained at least 00 times higher than the in vitro IC 99 in cervical tissue (3.3 ng/ ml). Local and systemic miconazole exposure was increased after coadministration with Ring-004 (.4 to 6-fold higher). Concomitant use of the two products was safe and well tolerated. One product-unrelated SAE occurred (acetabulum fracture); one Grade 2 event of vulvovaginal candidiasis (ring alone) was considered product-related. Conclusions: Concomitant use of Ring-004 and a single vaginal dose of miconazole nitrate altered the local and systemic levels of both drugs, but these changes are considered unlikely to affect adversely the efficacy of either drug. Daniel Sepúlveda-Crespo,2,3, María Jesús Serramía,2,3, Javier Sánchez-Rodríguez,2,3, Raquel Lorente,2,3, Rafael Gómez 3,4, Francisco Javier de la Mata 3,4, Jose Luis Jiménez 2, Mª Ángeles Muñoz-Fernández,2,3 Hospital General Universitario Gregorio Marañón, Laboratorio Inmunobiología Molecular, Madrid, Spain, 2 Hospital General Universitario Gregorio Marañón, Plataforma de Laboratorio, Madrid, Spain, 3 Networking Research Center of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain, 4 Universidad de Alcalá, Departamento de Química Inorgánica, Alcalá de Henares, Madrid, Spain Background: Self-administered topical microbicides may be very helpful tool for women and homosexuals to decrease new HIV infections. Polyanionic carbosilane dendrimers are considered HIV- entry inhibitors and antiretrovirals (ARVs) are the most advanced microbicides. Consequently, the combination approach should be taken into consideration when designing a new microbicide. In this regard, double o triple-combination of dendrimers and ARVs acting in the early stages of HIV- replication is an indispensable and beneficial tool in terms of efficacy and long-term safety in fighting the HIV/AIDS epidemic. We will show the latest results of the combination of two or three carbosilane dendrimers with tenofovir (TFV) and/or maraviroc (MRV) used as microbicides. Methods: Cytotoxicity in different cell lines in vitro, the anti-hiv- in TZM. bl cell line, vaginal irritation and subsequent histological analysis were showed. 48h post-infection inhibitory activity profile was determined by luciferase activity. Study of combined effects and the 50% effective concentration (EC 50 ) were performed using Calcusyn software. Results: Two- and three-drug combinations showed a greater broadspectrum anti-hiv- activity than the single-drugs, preserved this activity in acid environment or seminal fluid, and did not activate inflammatory response. The strongest combinations were with G2-STE6 dendrimer at constant ratios. They demonstrated strong synergistic activity profile due to the combination indices varied between 0.06 and Additionally, no irritation was detected in female mice after dendrimer vaginal administration. Conclusions: The two- and three-drug combination increases their antiviral potency and act synergistically as potential microbicide. Our results deserve especially further clinical research on dendrimer/ dendrimer, dendrimer/arvs, dendrimer/dendrimer/dendrimer, dendrimer/dendrimer/arvs, or dendrimer/arv/arv as microbicide against HIV

218 Posters Posters 5: Novel Formulations, Agents and Microbicides P5. More? Less? Just Right? The Role of Perceived Volume in Gel and Film Perceptibility During Intercourse, and its Impact on Product Preference P5.2 Vaginal Film User Evaluations: Developer Considerations from Initial Impressions and User Sensory Perceptions and Experiences during Vaginal Sex Kathleen M. Morrow, Rochelle K. Rosen 2, Sara Vargas 3, David Katz 4, Fava Joseph 3, Erna M. Kojic 5, David Friend 6, Lisa Rohan 7, Anthony Ham 8, Robert Buckheit 8 Miriam Hospital & Alpert Medical School of Brown University, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 2 Miriam Hospital & Brown University School of Public Health, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 3 Miriam Hospital, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 4 Duke University, Biomedical Engineering and Ob/Gyn, Durham, NC, United States, 5 Miriam Hospital & Alpert Medical School of Brown University, Division of Immunology, Providence, RI, United States, 6 CONRAD, Arlington, VA, United States, 7 Magee Womens Research Institute, Pittsburg, PA, United States, 8 ImQuest BioSciences, Inc., Frederick, MD, United States Background: We analyzed coital and pericoital vaginal perceptibility in 2 volumes of HEC gel (2 ml, 4 ml) and one x2 film to understand the role of volume and other properties in user sensory perceptions and experiences (USPEs) of vaginal formulations. Methods: 24 monogamous HIV/STI-negative heterosexual couples enrolled; 00% completed 3 product evaluation visits (random order). Women inserted product, then had vaginal sex with their male partner. All completed USPE surveys and 30 (5 couples) completed in-depth qualitative interviews (IDI) about each product s properties and performance. USPE scale scores, indicating ranges of sensations and experiences perceived by users, as well as qualitative thematic analyses, contribute to findings. Results: USPE scale scores showed significant differences in pairwise comparisons between each of 2 gel volumes and the film. Perceptibility (USPE) scores were lowest overall for film, and highest overall for 4mL gel. Volume differences were particularly relevant for: ) sensations at initial penetration; 2) ease of stroke, perceived wetness, and physical awareness of product during coitus; 3) perceived leakage and messiness; and 4) perceptions of product being sexually stimulating. IDIs described ranges of experiences with differences by volume and formulation. As expected, higher volume gel was generally considered to be the most lubricating and messiest. 2mL gel was perceived by some to be less messy and provide less lubrication, but this was not universal. Film was perceived as not adding volume; most said that it did not add lubrication. 44% of all participants (both genders) chose the 4mL gel as their preferred product; 29% chose the 2mL gel; 27% chose the film. Conclusions: Gel data indicate user-perceived differences by volume. Film data provides context showing the importance of the role of volume in product experience. While volume is important, user opinions of product performance for penetration, pleasure, and covert use are not determined by volume alone. Kathleen M. Morrow, Lisa Rohan 2, Rochelle K. Rosen 3, Joseph Fava 4, Anthony Ham 5, David Friend 6, David Katz 7, Erna M. Kojic 8, Robert Buckheit 5 Miriam Hospital & Alpert Medical School of Brown University, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 2 Magee Women s Research Institute, Pittsburg, PA, United States, 3 Miriam Hospital & Brown University School of Public Health, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 4 Miriam Hospital, Centers for Behavioral and Preventive Medicine, Providence, RI, United States, 5 ImQuest BioSciences, Inc., Frederick, MD, United States, 6 CONRAD, Arlington, VA, United States, 7 Duke University, Biomedical Engineering and Ob/Gyn, Durham, NC, United States, 8 Miriam Hospital & Alpert Medical School of Brown University, Division of Immunology, Providence, RI, United States Background: Drug delivery in prevention technology is critical, but is predicated on use adherence. The role of formulation characteristics/ properties of vaginal film (i.e., impressions of size, texture, color) and film perceptibility (user sensory perceptions/experiences) was explored during insertion and vaginal sex. Methods: 24 monogamous HIV-/STI- heterosexual couples enrolled in a mixed methods study. All (00% retention) completed 3 formulation evaluations, a placebo film. Female participants provided qualitative data regarding film attributes (size: x, x2, x3, 2x2-inch; texture: textured, smooth; color: clear, translucent, opaque) then inserted x2 film and had vaginal sex with their partner. Both partners completed USPE surveys and in-depth qualitative interviews. Results: Individual film characteristics were first rated separately, then a final preference for film design was given. Relatively equal numbers of women chose each film size. 3 of 23 women chose one smooth/one textured side. The majority chose translucent or opaque coloring. Data revealed a range of expectations about product performance. Some perceived the x films as difficult to insert and less efficacious, others felt the x film would insert easier, dissolve quicker, and result in less leakage. 2 x2 films elicited similar expectation ranges. USPE scale scores were low; women felt little-to-no lubrication, coating, leakage, etc. Design parameters called for a film that users would not be aware of and that would generate no leakage or messiness: these properties were achieved. However, low lubricity and coating sensations at initial penetration or during the first coital strokes were problematic, even causing a few couples to stop sex. Conclusions: The addition of vaginal film to the formulation parameter space for USPEs of vaginal products offers important considerations for developers. Given previous USPE analyses, initial lubrication may impact use adherence and thus is currently being explored in new film development. 28 HIV Research for Prevention 204 HIV R4P

219 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.3 Using Molecular Dynamics Techniques to Investigate the Influence of Glycans on the HIV- gp20 Envelope Protein Trimer Clint Mercuur, Natasha Wood, Simon Travers University of the Western Cape, South African National Bioinformatics Institute, Cape Town, South Africa P5.4 Development of a Temperature-recording Vaginal Ring for Monitoring User Adherence Peter Boyd, Clare McCoy, Diarmaid Murphy, Manjula Lusti- Narasimhan 2, Berglind Helgadóttir 3, Karl Malcolm Queen s University Belfast, School of Pharmacy, Belfast, United Kingdom, 2 World Health Organization (WHO), Geneva, Switzerland, 3 Star-Oddi Ltd., Gardabaer, Iceland Background: The gp20 glycoprotein present on the surface of the HIV virion is crucial for the recognition and binding to receptors on the host cell surface, as well as facilitation of the fusion of the viral envelope to the host cell membrane. The surface of gp20 is covered with N-linked glycans that can influence HIV- infectivity as well as affect the recognition of the virus by the host immune system, acting as a glycan shield from antibody recognition. Methods: Previous research has shown that the N-linked glycans bound to the surface of a gp20 monomer have a significant impact on the underlying dynamics of the protein. Here, we have expanded on this work by undertaking molecular dynamic modeling to explore the effect of N-linked glycans on the dynamics of the full gp20 trimer. Results: Our results illustrate the significance of the silent and active faces of the gp20 monomers, as well as showing the trimer-specific behavior of the glycans and glycan-protein interactions. Conclusions: Collectively our results provide a better understanding of the role that the glycan composition and distribution play during infection of a new cell. Background: Vaginal ring devices are being actively developed for controlled delivery of HIV microbicides and as multi-purpose prevention technology (MPT) products combining hormonal contraception with prevention of HIV and other sexually transmitted diseases. Presently, there is no reliable method for monitoring user adherence in HIV vaginal ring trials; previous acceptability studies have included some type of participant self-reporting mechanism, which have often been unreliable. More objective, quantitative and accurate methods for assessing adherence are needed. Methods: A silicone elastomer vaginal ring containing an encapsulated miniature temperature recording device has been developed that can capture and store real-time temperature data during the period of designated use. Devices were tested in both simulated vaginal environments and following vaginal placement in cynomolgus macaques. Various use protocols and data sampling rates were tested to simulate typical patient usage scenarios. Results: The temperature logging devices accurately recorded vaginal temperature in macaques, clearly showing the regular diurnal temperature cycle. When environmental temperature and vaginal temperature was significantly different, the device was able to accurately pinpoint the insertion and removal times. Based on the data collected it was possible to infer removal periods as short as 5 min when the external environmental temperature was 25 C. Accuracy increased with data sampling rate. Conclusions: This work provides proof-of-concept for monitoring adherence using a vaginal ring device containing an encapsulated temperature logger. The addition of one or more active agents into the ring body is not anticipated to affect the temperature monitoring function. A clinical study to compare self-reported user adherence data with that obtained by the device would be highly informative. 29

220 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.5 P5.6 Potential of RNA Aptamers in the Prevention of HIV- subtype C Infections Grace Mothepane London, Makobetsa Khati,2, Bongani Mayosi 2 CSIR, Biosciences, Pretoria, South Africa, 2 University of Cape Town, Medicine, Cape Town, South Africa Background: Compounds that have been used to prevent human immunodeficiency virus type-i (HIV-) infections include synthetic chemicals, plant extras and monoclonal antibodies. Although most of these compounds have potent antiviral activity, they often fail to progress to later stages of clinical trials due to high toxicity and lack of specificity. Therefore, as an alternative to circumvent the above mentioned limitations we used aptamers, which are small nucleic acid ligands that recognize their target with high specificity and have no toxicity in clinical applications. Methods: In this study, we evaluated efficacy of four gp20-aptamers against Env pseudovirus panel derived from HIV- subtype C, using virus inhibition assay in TZM-bl cells, as well as toxicity. Binding specificity of one potent aptamer (CSIR.) to gp20 was determined by Enzyme- Linked-Immunosorbent Assay. Subsequently, a virus inhibition assay was performed to test whether CSIR. can inhibit HIV- pseudotyped with vesicular stomatitis virus envelope glycoprotein (HIV-VSVG. Results: All four aptamers inhibited infectivity of 8-84 % of the tested viruses with mean inhibition concentration (IC 50 ) of nm. The specificity results showed that CSIR. only bound to gp20 and did not bind other tested proteins (HIV- gp4, mycobacterium tuberculosis virulent protein (CFP0), human interferon gamma (IFN-γ) and BSA). CSIR. also failed to inhibit HIV- pseudotyped with VSV-G protein and showed no toxicity in vitro, even at concentration (500 nm), which was 5 higher than one used for virus inhibition assays. Conclusions: Aptamers showed significant efficacy against HIV- subtype C isolates and specificity to gp20 without causing cytoxicity effects. These properties make aptamers attractive candidates for prevention of HIV-. Overcoming Phase Behavior and Surface Crystallization in a Tenofovir-levonorgestrel Intravaginal Ring Meredith Clark, Justin Clark 2, Todd Johnson 2, Namdev Shelke 2, David Friend, Patrick Kiser 3 CONRAD, Arlington, VA, United States, 2 University of Utah, Salt Lake City, UT, United States, 3 Northwestern University, Evanston, IL, United States Background: We have reported on the design of an intravaginal ring (IVR) engineered to deliver both tenofovir (TFV) and levonorgestrel (LNG). Because these drugs have divergent properties and release rates, sophisticated designs are required to deliver them in a controlled manner. These designs force us to push the materials science behind the IVR to the edge of what is possible. While exploring the physical chemistry of the IVR we discovered and solved a number of stability challenges that made the clinical development of this IVR possible. Methods: TFV/LNG IVR as previously described were subjected to various time-temperature profiles; drug release rates, presence of surface crystallization by polarized light microscopy, and polyurethane (PU) phase behavior by DSC were evaluated. Results: Rings made with 35wt% swelling PU and placed at 40 C for 0 and 60d went from 2±5 to 4± mg/d TFV due to changes in the effective diffusion coefficient of TFV in the PU with time. DSC showed a change in the water hydration state of the PU with time at 40 C with an exothermic peak at -50 C and enthalpy of -25 J/g. To compensate for this change we added a 4d annealing step at 40 C to the manufacturing process that allows the PU to achieve a more equilibrium conformation resulting in stable DSC and drug release profiles. Annealed rings made with a 60% swelling PU achieved steady-state release rates at 9±2 mg/d and eliminated the hydrated water enthalpy. LNG segments placed on storage showed a temperature dependent formation of LNG crystals on the surface due to supersaturation of drug in the PU with crystals forming after 30d at 40 C but not for at least yr at 4 C. Coating the LNG segment with glycerol or PVP, in which LNG has a low activity, inhibits LNG crystallization for >yr at 40 C. No LNG crystallization has been observed to date (5mo) on finished TFV/LNG IVR stored at 40 C. Conclusions: Quantitative evaluation of the phase behavior of the IVR allowed us to engineer an IVR not requiring cold chain storage. 220 HIV Research for Prevention 204 HIV R4P

221 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.7 Mass Transport Theory Improves Compartmental PK Modeling of Microbicides and Helps Guide Product Science and Development Yajing Gao, Andrew Yuan, David F. Katz,2 Duke University, Durham, NC, United States, 2 Duke University Medical Center, Obstetrics and Gynecology, Durham, NC, United States P5.8 Testing of Nanoparticle-based ARV Drug Combinations for Inhibiting Cell-free and Cellcell HIV Transmission Yonghou Jiang, Shijie Cao, Danielle Bright 2, Ian T. Sudyam 2, Kim A. Woodrow University of Washington, Department of Bioengineering, Seattle, WA, United States, 2 Seattle University, Department of Chemistry, Seattle, WA, United States Background: Understanding microbicide functionality, product design and evaluation derive from synergy of experimental (in vitro, in vivo) and computational (modeling) analyses. Modeling can deconstruct and elucidate many elements of cause and effect in this complex, multivariate process. Its potential has not been fully realized in the science and development of anti-hiv strategies, e.g. in pharmacokinetic analysis. Mechanistic modeling of microbicide PK processes - for different vehicles (e.g. gels, rings) and different types of drugs - can provide a framework to: () identify salient properties of vehicle, drug, host environment and dose regimen that govern PK; (2) input those properties to make quantitative comparisons and predictions that help design and interpret in vivo studies in humans and animals, and in vitro studies of drug release/transport and HIV neutralization. Methods: Principles of mass transport theory were applied to vaginal delivery of microbicide drugs by clinical and prototype gels and rings. Compartments in the models were vehicle, lumenal fluids, epithelium, stroma and blood stream. Diffusion and convection were primary mechanisms of drug transport. Systems of coupled partial differential equations were derived/solved, outputting drug concentrations vs. time/ location in each compartment. Results: Results include tradeoffs in gel volume (2-4mL) and loading (0.5-.5%) for Tenofovir, IQP0528 and other drugs in achieving mucosal concentrations that reach prophylactic (e.g. EC50) levels, showing how frequency of dosing can sustain these levels. IVR geometry and release flux history are related to such target concentrations, including effects of transient ring removal. Scaling rules for human vs. animal dosing are given. Conclusions: Mechanistic modeling fills gaps in microbicide drug delivery science, helping define and organize effects of the many factors involved. This knowledge translates to methodology for use in rational product design and performance evaluation, in vitro and in vivo. Background: Strategies that enable ARV drugs to be easily combined and provide sustained antiretroviral activity have the greatest potential to impact the efficacy of next generation topical microbicides. To overcome challenges associated with formulating multiple ARV compounds that are chemically incompatible, nanoparticles were fabricated to encapsulate individual ARV drugs that are then delivered in combination. We show that novel combinations of ARV-formulated nanoparticles that have activity against cell-cell as well as cell-free virus transmission. Methods: ARV drug-nanoparticles were prepared using emulsion-solvent evaporation techniques to incorporate maraviroc (MVC), etravirine (ETR), and raltegravir (RAL) into PLGA nanoparticles. We compared the antiviral potency of the free and formulated drug combinations for all three pairwise combinations of ETR, RAL and MVC against both cellfree and cell-associated HIV- infection in vitro. The efficacy of ARVdrug nanoparticle combinations was also assessed in a macaque cervicovaginal explant model using RT-SHIV. Results: We observed that ARV-drug nanoparticle combinations that include ETR exhibit significant antiviral potency and dose-reduction against both cell free and cell-associated HIV- BaL infection in vitro. ETR-NP combined with MVC-NP or RAL-NP showed an approximately 0- fold dose reduction compared to the similar unformulated combinations against cell-free virus infection. We also observed that cell-cell HIV- BaL transmission was inhibited at least 0-fold more effectively with drug combinations that included ETR compared to combinations of MVC and RAL. ETR-NP combined with MVC-NP or RAL-NP showed approx. threetimes greater potency compared to the unformulated combinations of the same ARV drugs. Conclusions: We expect that the versatility of nanoparticle delivery platforms will result in broad applications for HIV chemoprophylaxis, particularly via routes with limited dosage forms for combination drug delivery to the genital and rectal mucosa. 22

222 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.9 DuoGel : The Development of A Multi-drug Dual Chamber Vaginal/Rectal Anti-HIV Microbicide Gel Anthony S. Ham, Sean T. Nugent, Jennifer J. Peters 2, David F. Katz 2, Cory Shelter 3, Charlene S. Dezzutti 3, Ashlee D. Boczar, Karen W. Buckheit, Robert W. Buckheit Jr. ImQuest BioSciences, Frederick, MD, United States, 2 Duke University, Durham, NC, United States, 3 Magee Women s Research Institute, Pittsburgh, PA, United States P5.20 Imaging and Visual Evaluation for the Confirmation of Vaginal Film Spreading and Retention Sheila Grab,2, Yvonne Cosgrove Sweeney 3, Dorothy Patton 3, Lisa Rohan,2 University of Pittsburgh, Pharmaceutical Sciences, School of Pharmacy, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 3 University of Washington, Obstetrics and Gynecology, Seattle, WA, United States Background: The DuoGel, a multi-drug anti-hiv gel microbicide, is a single product for safe and effective dual chamber administration which has been developed to address the high incidence of both vaginal and anal intercourse in the same sexual act. The DuoGel overcomes significant safety issues involved with the rectal use of vaginally designed gels, reduces the complexity of maintaining separate dosage forms, and addresses the need for prevention products in the MSM population. Methods: The DuoGel containing IQP-0528 and Tenofovir was formulated from GRAS excipients approved for both vaginal and rectal administration. The ph and osmolality was defined by a target product profile. Viscosity, rheological spreading, and gel distribution were measured under various in vitro shear conditions. Ex vivo drug release was performed in Franz cells into full thickness vaginal tissue over 6 hours. In vitro toxicity was performed against established cell lines and Lactobacilli for 24 hours. In vitro efficacy was performed in PBMCs against HIV- infection for 7 days. The ex vivo toxicity, and efficacy was performed in both polarized explant ectocervical and colorectal tissues. Results: The DuoGel formulation was developed to a specific ph (6.00) and osmolality (350 mmol/kg) to accommodate rectal administration. With a viscosity of 4.8 ± 5.73 Pa*s at s -, a gel distribution of 87.8 cm 2 (60% universal placebo) was measured. The DuoGel produced an in vitro and ex vivo drug release rate of 0.48 ± 0.07 µg/cm 2 hr for IQP and 34 ± 3.8 µg/cm 2 hr for Tenofovir to prevent HIV- infection in both vaginal and rectal environments with an EC 50 value of 0.48 ± ng/ml. No in vitro cellular or bacterial toxicity up to a high concentration of 000 µg/ml and no loss in cellular tissue viability in both explant ectocervical and colorectal tissue was measured. Conclusions: This study has developed a multi-drug DuoGel formulation that has the potential to safely prevent HIV- transmission in both the vagina and rectum. Background: The vaginal thin film is being evaluated as a drug delivery platform for topical pre-exposure prophylaxis of HIV, but there is little information regarding film distribution and drug release kinetics within the vagina. The objective of this study was to evaluate the impact of film volume on tenofovir (TFV) drug release using in vitro mechanical testing, dissolution testing and visual monitoring of product retention in a macaque model. MRI imaging was conducted to study product distribution in this model. Methods: Solvent cast vaginal films containing TFV and/or gadobenate dimeglumine (GD-contrast agent for MRI imaging) were characterized for mass, thickness, drug and water content, puncture strength and disintegration. Films of various thicknesses were manufactured with a blue dye and inserted intravaginally in 4 pigtail macaques for visual retention studies using colposcopy. Films were inserted intravaginally in 4 pigtail macaques using MRI to track film dispersal. Results: Films contained an average of 8.44 mg of TFV. In vitro drug release rates (mg/ time) were 4.56 and.65 for the 90 and 500µm films, respectively. Visual film retention observations in the macaque showed that increased film thickness resulted in a longer vagina film residence time. MRI studies in this same model demonstrated that GD contrast was clearly seen throughout the vagina and ectocervix at the initial time point which was 4 hours post film placement. Conclusions: In vivo, the vaginal film has been shown to effectively coat the vaginal and ectocervical tissues without migrating to upper reproductive tract (URT) or peritoneal spaces. Changes in film volume can affect drug release profile and vagina film residence time. Data obtained suggest that the film platform may provide sustained drug release offering a wider time frame of protection against HIV. 222 HIV Research for Prevention 204 HIV R4P

223 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.2 Transient Protein Expression Facilitates X-ray Structural Studies of HIV- Alex Druz, Marie Pancera, Priyamvada Acharya, Tongqing Zhou, Gordon Joyce, Adi Ofek, Peter D. Kwong NIAID, National Institutes of Health, Vaccine Research Center, Bethesda, MD, United States P5.22 Fourteen-day Safety of Daily Vaginal Administration of Dapivirine-loaded Nanoparticles in a Mouse Model José das Neves,2, Rute Nunes 2, Bruno Sarmento,2 IINFACTS, Instituto Superior de Ciências da Saúde-Norte, CESPU, Gandra, Portugal, 2 INEB Instituto de Engenharia Biomédica, Porto, Portugal Background: Transient protein expression is a method of choice to produce recombinant HIV- proteins for the quick assessment of their structural characteristics. Large-scale transient protein expression in mammalian systems such as HEK293 or CHO cells allows obtaining milligram quantities of eukaryotic viral proteins and antibodies with a relatively high-throughput which require accurate post-translational modifications for proper folding and/or a part of large multimeric complexes. Methods: We developed the large-scale transient protein production platform in HEK293 cells for rapid production of the milligram quantities of soluble HIV- immunogens and monoclonal antibodies sufficient to facilitate the structural studies of HIV- sites of vulnerability and antibodies. Results: Large-scale transient transfection platform provided the means to obtain mg quantities of HIV- immunogens and antibodies with high degree of variability which allowed us to perform X-ray structural characterization of HIV- viral spike and its sites of vulnerability. Crystal structures of soluble HIV- envelope proteins provided atomiclevel details on functional constraints, conformational flexibility and antibodies binding sites. Conclusions: Rapid and efficient transient production of a number of difficult-to-express HIV- envelope proteins and antibodies not only facilitates HIV- structural studies to delineate the atomic level details, flexibility and conformational constraints of the viral spike but also allows to design and assess structurally stabilized HIV- immunogens which altogether establishes the integrative approach en route to vaccine development. Background: We previously showed the ability of poly(ethylene oxide)- modified polycaprolactone nanoparticles (PEO-PCL NPs) loaded with dapivirine to modulate genital pharmacokinetics of the drug after vaginal delivery in mice. In this study we evaluated the safety of NPs in the same model. Methods: Dapivirine-loaded NPs at a drug concentration of 0.02% in PBS were administered intravaginally to ICR mice once-daily for 4 days. Daily vaginal washing was performed immediately before each administration and lavages assayed for IL-β, IL-6, KC and MIP- α content. Animals were sacrificed 24h after the last administration, and genital tissues/selected organs collected, processed for H&E and analyzed for histology. Results were compared to those of animals similarly treated with PBS, 0.02% dapivirine or 2% nonoxynol-9 (N- 9), and untreated mice. Further, drug accumulation in lavages, blood plasma and tissues/organs was assessed by HPLC-UV. Results: No gross changes were observed during necropsy. Histological evaluation of the genital tract (vagina, uterus, ovaries) showed no changes in animals treated with NPs, dapivirine or PBS. No accumulation or presence of particulate matter (NPs/drug) was detected. No to mild drug accumulation was observed in lavages/blood plasma/organs. Results were comparable to those of untreated animals and contrasting with marked vaginal and uterine epithelial damage in the N-9 group. No systemic histological changes were observed. Analysis of cytokine/ chemokine content in lavages revealed no differences in animals treated with NPs or drug suspension, as compared to the PBS group; small decreases in KC and IL-β levels during the first -2 days, as compared to untreated mice, were associated with daily vaginal washing. N-9 use induced the increase of all cytokines/chemokines after -4 consecutive administrations. Conclusions: Obtained data support that dapivirine-loaded PEO-PCL NPs are safe upon once-daily vaginal administration for 4 days in a mouse model

224 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.23 Development of a DuoGel for Vaginal and Rectal Delivery of Microbicide Products Karen Buckheit, Ashlee D. Boczar, Caitlin A. Buchholz, Sean Nugent, Anthony S. Ham, Cory Shetler 2, Charlene S. Dezzutti 2, Robert W. Buckheit Jr. ImQuest BioSciences Inc., Frederick, MD, United States, 2 University of Pittsburgh Medical Center and Magee-Women Research Institute, Pittsburgh, PA, United States P5.24 Female Sex Hormone Regulation of Tenofovir-diphosphate in Human Female Reproductive Tract (FRT) Cells in Culture Zheng Shen, John V. Fahey, Marta Rodriguez-Garcia, Jack E. Bodwell, Angela D.M. Kashuba 2, Charles R. Wira Geisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United States, 2 UNC Eshelman School of Pharmacy, The Division of Pharmacotherapy and Experimental Therapeutics, Chapel Hill, NC, United States Background: It has been over 30 years since the first cases of HIV/ AIDS and prevention of infection through vaccination remains elusive. Thus, alternative products, technologies and strategies have emerged, including pre-exposure prophylaxis through oral ARVs and vaginal and rectal delivery of topical microbicides to prevent sexual transmission of HIV. It is well understood that men and women engage in receptive anal intercourse and women engage in both vaginal and anal intercourse during the same sexual encounter, creating a strong rationale for the development of microbicide delivery strategies targeting both vaginal and rectal routes of transmission in order to reduce HIV transmission. ImQuest has developed the NNRTI IQP-0528 as a vaginal gel based on its potency, multiple mechanisms of antiviral action, and its ability to act in an additive or synergistic manner with other microbicide products. We have continued the development of the microbicide IQP-0528 with a new focus on a product which safely delivers this single agent to both compartments (DuoGel) as well as a combination DuoGel comprised of IQP-0528 and tenofovir. Methods: Based on the IQP-0528 vaginal gel and a defined product profile for a DuoGel, a series of single-agent and combination DuoGels having the defined osmolality, ph and viscoelastic properties to be safely used in both the vagina and rectum have been produced. Utilizing well established in vitro and ex vivo microbicide development assays we assessed the efficacy and toxicity of each of the placebo and APIcontaining gels. Results: The microbicide activity of these gels was not affected by the presence of vaginal and seminal simulants. No toxicity of the gels was observed to representative cell lines, epivaginal tissue, the normal flora Lactobacillus, or cervical or colorectal explant tissue. Conclusions: Based on these biological data a single agent DuoGel is being advanced to an IND and human clinical testing and a combination IQP-0528/Tenofovir DuoGel is being developed. Background: The conflicting results of recent pre-exposure prophylaxis (PrEP) trials utilizing tenofovir (TFV) for the prevention of HIV infection in women led us to evaluate intracellular TFV-diphosphate (TFV-DP) in cells from the FRT and whether estradiol (E 2 ) and progesterone (P4) influence the presence of TFV-DP in these cells. Methods: FRT tissues were obtained following hysterectomy for benign reasons from HIV-negative women. Epithelial cells (EC), fibroblasts and CD4 + T cells from the ectocervix (ECX) endocervix (CX) endometrium (EM) were isolated by enzymatic digestion and/or magnetic bead purification. Cells were treated with TFV for 24hr and intracellular TFV- DP measured by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results: We found that TFV-DP concentrations vary significantly with the cell type analyzed and the site in the FRT. EM-EC had 2-fold higher TFV-DP than EC from the CX and ECX. TFV-DP in fibroblasts from EM, CX and ECX were comparable. Concentrations of TFV-DP in EC were ~5-fold greater than that seen in fibroblasts. In studies with E 2 and P4 added in culture to FRT cells, E 2 increased TFV-DP in EM, CX/ECX epithelial cells, but had no effect on fibroblasts or CD4 + T cells from FRT tissues. In contrast, P4 alone and in combination with E 2 decreased TFV-DP concentrations in FRT CD4 + T cells. Conclusions: These results demonstrate that TFV-DP is produced by multiple cell types throughout the FRT and that intracellular concentrations vary with the cell type analyzed. Our results suggest that TFV-DP concentrations in epithelial cells and fibroblasts may either be a repository for TFV-DP, which when converted to TFV is available for protection of CD4 + T cells and macrophages, or a potential sink, which interferes with TFV-DP target cell availability. Further, these results indicate that E 2 and/or P4 regulate the intracellular concentrations of TFV-DP in epithelial cells and CD4 + T cells and that intracellular TFV-DP varies with cell type and location in the FRT. 224 HIV Research for Prevention 204 HIV R4P

225 Posters 5: Novel Formulations, Agents and Microbicides Wednesday, 29 October P5.25 Enhancing the Solubility of HIV--neutralizing Antibody 0E8 Young Do Kwon, Ivelin S. Georgiev, Baoshan Zhang, Krisha McKee, Sijy O Dell, Alex Druz, Wei Shi, Mark Connors 2, John R. Mascola, Peter D. Kwong Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 2 Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States P5.26 LB Exploring Innovative Approaches to the Formulation of Microbicides to Boost Antiretroviral Drug Delivery and Activity at Mucosal Sites Julia N. Ekeruche-Makinde, Abbey Evans, Carolina Herrera, Kelly Charles 2, Robin Shattock Imperial College London, Department of Medicine, London, United Kingdom, 2 King s College London, London, United Kingdom Background: The human monoclonal antibody, 0E8, targets the membrane-proximal external region of gp4 on HIV- Env and neutralizes ~98% of HIV- isolates. However, it is prone to aggregation at neutral ph and this less-than-ideal solubility poses a challenge to the development of 0E8 as a prophylactic agent for passive protection from HIV- infections in humans. Methods: We used somatic variants and employed structural and computational approaches to enhance the solubility of 0E8, which we assessed by measuring the turbidity of variants in phosphate buffered saline. Neutralization potency was measured via luciferase/tzm-bl cellbased assay on a 9-virus panel. Results: We identified a somatic variant of the heavy chain 0E8 (named HC6), which slightly improved neutralization potency relative to 0E8, while displaying similar breadth and solubility. We altered four hydrophobic residues in the framework 3 region of HC6 and found that this alteration (L72D, I75K, F77T, and M84T) improved solubility by ~0- fold. When we introduced these mutations into another heavy chain variant of 0E8, H6dN52, solubility increased even further (>20-fold relative to 0E8); however, this variant neutralized the selected viruses over 0-fold more weakly than 0E8. Mapping sequence differences between HC6 and H6dN52 suggested that this reduced potency might relate to changes in four residues in close proximity to the gp4 epitope: D28, N3, T52 and Y98. Incorporation of changes at these residues into H6dN52 (L72D, S74T, I75K, and F77T) increased potency to a level (geometric mean IC 50 =0.68 ug/ml), which was comparable to that of 0E8, while retaining a >20-fold increase in solubility. Conclusions: By combining a structure-based approach and the natural variation in potency and solubility for somatic variants of 0E8, we successfully engineered a variant of 0E8 with solubility improved by more than 20-fold, while retaining its impressive neutralization breadth and potency. Background: In the absence of a vaccine against HIV infection topically applied pre-exposure prophylaxis with microbicides represents one of the promising options for protection in women who have no choice in their partners use of condoms. When delivered at the point of entry microbicides work to deliver localised concentrations of drugs that have the potential to block the earliest stages of infection and prevent transmission. The effectiveness of microbicides as a preventative strategy has been demonstrated in humans and non-human primates. The use of formulated combinations of antiretroviral drugs as microbicides has been promoted as a means to check viral escape and provide broad protection. Moreover it has been suggested that a number of antiretroviral drugs may be capable of interacting with cellular mechanisms that serve to boost drug activity at mucosal sites. Methods: As part of an EU funded project we investigated the possibility of developing optimized microbicide formulations for vaginal and rectal delivery. We examined the advantages of double and triple combination approaches to the inhibition of virus entry and transmission in cells derived from human cervical tissue explants, peripheral blood mononuclear cells and dendritic cells. Results: In the models tested we obtained evidence supporting the use of two reverse transcriptase inhibitors (Tenofovir and TMC-20) and a protease inhibitor (Darunavir) in combination to inhibit infection and transmission of HIV-. This effect was demonstrated in a fold reduction in the IC 50 values of the drugs in combination when compared with single drugs. We then showed the effect of the drugs on the ATP activity of the membrane bound cellular transporter p-glycoprotein. Conclusions: Our results highlight the potential to boost intracellular concentrations and the antiviral effect of the drugs by modulating the activity of cell transporter molecules. Subsequent experiments will seek to examine this outcome in human cellular and mucosal tissue explant models

226 Posters Posters 5: Novel Formulations, Agents and Microbicides P5.27 LB First-in-Human Safety and Pharmacokinetics (PK) of a MIV-50/Zinc Acetate/Carrageenan Gel (PC-005) George Creasy, Craig Hoesley 2, Barbara Friedland 3, Shimin Zhang, Marlena Plagianos, Kyle Kleinbeck, Keith Levendosky, Jose Fernández-Romero, Tom Zydowsky Population Council, Center for Biomedical Research, New York, NY, United States, 2 University of Alabama at Birmingham, Department of Medicine, Division of Infectious Diseases, Birmingham, AL, United States, 3 Population Council, New York, NY, United States P5.28 LB Structure of BMS-806, a Small-molecule HIV- Entry Inhibitor, Bound to BG505 SOSIP.664 HIV- Env Trimer Marie Pancera, Aliaksandr Druz, Tongqing Zhou, Sijy O Dell, Mark Louder, Navid Madani 2, Alon Herschhorn 2, Joseph Sodroski 2, John R. Mascola, Peter D. Kwong Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 2 Dana Farber Cancer Institute, Boston, MA, United States Background: The candidate microbicide, PC-005, completely protects Depo-Provera-treated macaques from a single vaginal SHIV-RT challenge 8h post dose, and significantly reduces HPV and HSV-2 infection in murine models. PC-005 contains 50µM MIV-50 (NNRTI) and 4mM zinc acetate dihydrate in a carrageenan gel. Methods: In preparation for a Phase trial, an open-label, safety run-in was conducted at the University of Alabama at Birmingham to assess the safety and pharmacokinetics (PK) of PC-005. Healthy, sexuallyabstinent, HIV and Hepatitis B/C negative, STI-free, non-pregnant women aged 9-49 on effective contraception, were eligible. Under clinical supervision, women inserted 4ml of PC-005 once daily for 3 days. Evaluations included physical exam, pelvic exam with colposcopy, EKG, vitals, and safety labs. Blood was drawn for PK assessment at 0.5,, 2, 3, 4, 6, 8, 0, 2, 4, 6, 20, and 24h post-doses and 3; and 48h and 72h post-dose 3. Results: From June-July 204, 5 women (2 Black, 2 White, Native American) completed the study (3 doses). Median age was 29 (range 29-38). Three women reported 4 possibly-related AEs; 3 were DAIDS Grade : vaginal discharge, intermenstrual bleeding, lower abdominal pressure; was DAIDS Grade 2: vaginal itching. Safety labs, physical exams, vital signs, and colposcopy were all normal or not considered clinically significant by investigators. All subjects had detectable MIV- 50 blood levels after dosing; no accumulation was noted. On Day 3, the median (and range) of MIV-50 PK parameters were: T /2 of 4.98h ( ); C max of 77.4 pg/ml ( ); T max of 4h (2-6); AUC last of pg h/ml ( ); AUC inf of pg h/ml ( ). There was no increase in zinc blood levels from baseline; median C max of zinc was 79 µg/ml (58-94) on Day 3. Conclusions: PC-005 was well tolerated after 3 days of dosing in 5 healthy women. MIV-50 was absorbed with low levels observed systemically; no accumulation was noted. Zinc levels were unchanged from baseline. Background: The HIV--envelope (Env) spike is a conformational machine that switches between prefusion and postfusion conformations to facilitate HIV- entry. Extensive interest has focused on the prefusion mature closed conformation, as it is the target of neutralizing antibodies. One hall-mark of this conformation is the recognition by the small molecule entry inhibitor, BMS-806, which is currently in preclinical trial. Mutagenesis has shown that BMS-806 binds to specific residues within the CD4-binding pocket of Env. The mechanism of BMS-806 remains unclear. Methods: We obtained a lattice of HIV- Env, bound by antibodies 35O22 and PGT22 that diffracts sufficiently to allow chemical details to be visualized by X-ray crystallography. By using this lattice, we were able to visualize how BMS-806 binds to HIV- Env in the context of a prefusion closed mature state using BG505 SOSIP.664. Results: Structure of BMS-806 bound to BG505 SOSIP.664 will be presented and comparison to unbound structure will reveal BMS- 806 mechanism. Conclusions: We obtained a structure of BMS-806, a potent entry inhibitor, bound to HIV- Env trimer. Binding of BMS-806 to the 35O22/PGT22-bound trimer provides additional evidence for the functional validity of this antibody-bound structure. Moreover, the chemical details of BMS-806 interaction should allow for its structurebased enhancement. 226 HIV Research for Prevention 204 HIV R4P

227 Posters 6: Passive Antibody Functions P6.0 gp20/cd4 Blocking Antibodies Are Frequently Elicited in ART-naïve Chronically HIV- Infected Individuals Wednesday, 29 October P6.02 Sequence Characteristics of HIV- B Envelope Proteins and its Potential Correlation with Broadly Neutralizing Activity Jorge Carrillo,2, Luis M Molinos, Maria Luisa Rodriguez de la Concepción, Silvia Marfil, Elisabet García, Bonaventura Clotet 3,4, Juliá Blanco 5,6 AIDS Research Institute IrsiCaixa, Badalona, Spain, 2 Centro de Investigación en Salud Internacional de Barcelona (CRESIB), Barcelona, Spain, 3 AIDS Research Institute IrsiCaixa, IGTP, Universidad Autónoma de Barcelona, Badalona, Spain, 4 Fundació Lluita contra la SIDA, Badalona, Spain, 5 AIDS Research Institute Irsicaixa, IGTP, Universidad Autónoma de Barcelona, Badalona, Spain, 6 Universidad de VIC, Vic, Spain Background: Antibodies (Abs) with the ability to block the interaction of gp20/cd4, such as anti-cd4bs Abs, can prevent from infection by HIV- and their elicitation may be an interesting goal for any vaccination strategy. However, the in vivo quantification of these immunoglobulins (Igs) remains challenging mainly because they recognize conformational epitopes, which are difficult to mimic in vitro. Methods: The presence of gp20/cd4 blocking Abs in plasma samples from 36 ART-naïve Chronic HIV- infected patients and 0 uninfected healthy controls was evaluated by a competitive flow cytometry assay using a hucd4/murine-igg fusion protein. Anti-CD4bs Abs were quantified by ELISA using resurfaces proteins RSC3 and RSC3Δ37I. Neutralization activity of plasma samples was determined by a TZM-blbased neutralization assay. Results: 35 out of 36 HIV- infected patients showed CD4/gp20 blocking Abs. No correlation with anti-cd4bs ELISA data was observed suggesting that not all anti-cd4bs Abs showed gp20/cd4 blocking capacity. In addition, 0 out of 36 plasma samples lacking anti-cd4bs Abs in ELISA assays were able to block the interaction between gp20 and its receptor indicating that Abs recognizing other epitopes can play a similar role. Although a poor correlation between gp20/cd4 blocking Abs and viral load was established, these Igs were developed mainly in patients with viral load higher than 5000 copies/ml and maintained at least during one year. These Abs correlated with the neutralizing capacity of plasma samples suggesting that they can contribute to the neutralizing workforce of plasma. Conclusions: The frequency of Abs capable of blocking the interaction between CD4 and gp20 in HIV- infected patients was higher than previously expected, bringing together several specificities such as the CD4bs but also other undefined epitopes. The characterization of this CD4/gp20 blocking response may provide valuable information for the design of a successful vaccine. Yanli Chen, Yuanyuan Hu, Zhenpeng Li, Li Ren, Liying Ma, Yuhua Ruan, Kunxue Hong, Yiming Shao National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China Background: Current HIV- vaccine strategies fail to elicit broadly neutralizing antibodies(bnabs). Further understanding of the mechanisms of induction of BnAbs in natural HIV- infection will provide insights on improving the current vaccine strategies. The aim of this study is to identify samples with BnAbs activities from HIV- B infected individuals and to explore potential sequence characteristics of env genes which correlate with broadly neutralizing activity. Methods: Using a panel of 25 Env-pseudoviruses including HIV- clade B, C, A, CRF07_BC and CRF0_AE strains, six samples with BnAbs activities were identified from 03 samples with chronic HIV- B infection. 64 env sequences were amplified by single genome amplification assay from these six samples and sequence characteristics were analyzed. Results: Each of these six samples with BnAbs activities was able to neutralize more than 80% test strains, in which the neutralizing breadth for clade B strains is the biggest. Sequence analysis indicated the genetic diversity mainly lied in VV2, V4, and V5 sub-regions; env genes from these samples have a longer length and more glycosylation sites at VV2,V4 regions, but less net charge at V3 loop; preliminary sequence analysis demonstrated env genes from these samples have escape mutations in key sites of 0E8, 2G2, PGT27/28 and PG6/ PG9 epitopes, and the proportion was 77.8%, 30.6%, 27.8% and 3.9% respectively. In addition, 46 specific sites and 8 co-variation patterns were identified in env genes, which may be associated with broadly neutralization activity. Conclusions: This study demonstrated that a relatively high prevalence of BnAbs responses was detected in chronic HIV- B infection. Sequence analysis suggested that HIV- may escape neutralization via increasing length and the glycosylation of VV2 and V4 loop and reducing V3 ring static charge, and different epitomes of HIV- env genes from samples with BnAbs activities endure varying immune pressures

228 Posters Posters 6: Passive Antibody Functions P6.03 Investigating Broad Neutralization in HIV- Non-B Subtype Infection in Yaoundé, Cameroon P6.04 Anti-MPER Antibodies with Heterogeneous Neutralization Capacity Are Detectable in Most Untreated HIV Infected Individuals Colleen Courtney,2, Luzia Mayr 2, Lilian Nogueira 3, Johnson Ngai 4, Michael Seaman 5, Florian Klein 3, Michel Nussenzweig 3, Phillipe Nyambi 2, Susan Zolla-Pazner 2 New York University School of Medicine, Microbiology, New York, NY, United States, 2 Veterans Affairs Medical Center, Pathology, New York, NY, United States, 3 The Rockefeller University, New York, NY, United States, 4 Medical Diagnostic Center, Yaounde, Cameroon, 5 Beth Israel Deaconess Medical Center, Boston, MA, United States Background: A hallmark of HIV is its variability, which within group M viruses includes 9 subtypes and over 50 circulating recombinant forms (CRF)s. Cameroon is an epicenter of the disease known for circulation of many subtypes and CRFs, which provides a unique setting to study the immune response to HIV. It is crucial for a successful vaccine to elicit an adaptive immune response against a broad range of viral subtypes. Methods: We have performed subtype analysis of viral RNA isolated from the plasma of 63 HIV- infected Cameroonian subjects to verify non-b subtype infection and to monitor for recombination. From the non-b, drug naïve cohort, we have isolated IgG (N=264) to screen for neutralization using a tier 2, pseudovirus panel including subtypes A, B, C, and 0_AE. Samples capable of neutralizing multiple pseudoviruses at high potency were selected for further study to determine neutralization capacity against an extended panel and their ability to bind gp60 envelope peptides. Results: Subtypes A, D, F, G, and K were identified as well as many recombinant forms including the predominant subtype CRF02_AG. The neutralization screen concluded that 9.5% of the IgG samples were able to neutralize at least 2 pseudoviruses at low IgG concentrations. Neutralization of the extended panel revealed that these IgG samples were also able to neutralize 02_AG,G, and AC subtypes. We found that all of the broadly neutralizing IgG samples bound envelope protein gp4, over 80% bound gp20 proteins, 79% bound a scaffolded VV2 peptide, and 00% of the samples bound cyclic V3 constructs. Conclusions: We have taken the first look at broad neutralization in a cohort infected with non-b subtype viruses and numerous CRFs. IgG from these patients is capable of neutralizing pseudoviruses across clades with high breadth and potency. We have shown that these samples bind strongly to envelope peptides and hope to continue to elucidate how these antibodies are conferring neutralization in order to guide future vaccine design. Luis M. Molinos-Albert, Jorge Carrillo, Marta Curriu, Maria L. Rodriguez de la Concepción, Silvia Marfil, Elisabeth García, Bonaventura Clotet, Julià Blanco IrsiCaixa AIDS Research Institute-HIVACAT, Badalona, Spain Background: The Membrane Proximal External Region (MPER) of the HIV- envelope glycoprotein gp4 is targeted by broadly neutralizing antibodies and constitutes, thus, a promising target for a preventive vaccine. However, the localization of this epitope in a hydrophobic environment seems to hamper the elicitation of these antibodies in HIV infected individuals. Methods: We have quantified and characterized MPER antibodies by ELISA and by flow cytometry using a collection of mini gp4-derived proteins expressed on the surface of 293T cells. Longitudinal plasma samples from 35 HIV infected individuals were assayed for MPER recognition and MPER dependent neutralizing capacity using HIV-2 viruses engrafted with HIV- MPER sequences. Results: Miniproteins devoid of the cysteine loop of gp4 exposed the MPER on 293T cell membrane. Anti-MPER antibodies were identified in most individuals and were stable when analyzed in longitudinal samples. The magnitude of the responses was strongly correlated with the global response to the HIV Env glycoprotein, suggesting no specific limitation for anti-mper antibodies. Peptide mapping showed poor recognition of C-terminal MPER moiety and a wide presence of antibodies against the 2F5 epitope. However, antibody titers failed to correlate with 2F5- blocking activity and, more importantly, with the specific neutralization of HIV-2 chimeric viruses bearing the HIV- MPER sequence; suggesting a strong functional heterogeneity in anti-mper humoral responses. Conclusions: Anti-MPER antibodies can be detected in the vast majority of HIV infected individuals and are generated in the context of the global anti-env response. However, the neutralizing capacity is heterogeneous suggesting that eliciting neutralizing anti-mper antibodies by immunization might require refinement of immunogens to skip nonneutralizing responses. 228 HIV Research for Prevention 204 HIV R4P

229 Posters 6: Passive Antibody Functions P6.05 Wednesday, 29 October P6.06 Antibodies Induced Lysis of Primary Infected Cells by ADCC Is HIV- Strain Specific Marina Biedma, Thomas Decoville, Bin Su, Sylvie Schmidt 2, Géraldine Laumond 2, Christiane Moog INSERM U 09, FMTS, Strasbourg, France, 2 INSERM U 0, FMTS, Strasbourg, France Background: Understanding immune responses aiming to control or prevent HIV- infection is fundamental for the design of vaccine strategies. Increasing evidences suggest that antibodies (Abs) bound to innate immune effector cells may play a role in protection from infection. Antibody-dependent cellular cytotoxicity (ADCC) mediated by Natural Killer (NK) cells is a complex process involving Abs that bridge infected target cells with FcγRIII bearing NK cells. This binding results in cell lysis, besides direct lysis as part of innate NK cell-mediated mechanism. In this study, the antiviral contribution of ADCC was investigated using physiologically relevant conditions on primary HIV- infected CD4T cells and using autologous primary NK cells. Methods: We infected CD4 + enriched T-cells with different HIV- R5 isolates to record 20% infected cells. Autologous NK cells were at an effector to target ratio of :. To get rid of direct lysis of infected cells by NK, the percentage of infected cells was determined by flow cytometry after 4 h incubation in the presence (ADCC) or in the absence (direct lysis) of Abs. Results: We found that effector NK cells display potent reduction of HIV- infected target cells by polyclonal and monoclonal Abs beyond the direct lysis observed in the absence of Abs. Interestingly, the extent of ADCC was highly dependent on the virus strain used suggesting that vaccine should induce Abs with broad ADCC as for Ab neutralization. Moreover, the percentage of cell lysis recorded was different to that obtained by analyzing surrogate marker of ADCC in vitro as NK CD07a expression levels or measurement of effector molecules delivered into target cells. Conclusions: Detection of lysis of primary infected cells support the potential in vivo role of ADCC as additional inhibitory mechanism involved in protection and highlight the necessity to further compare the currently used ADCC assays for their physiological relevance in protection against HIV-. V2/C2 Region of HIV- Clade C Primary Envelopes Confer Altered Neutralization Susceptibilities to IgGb2 and PG9 Monoclonal Antibodies Shilpa Patil, Ipsita Choudhary 2, Rajesh Ringe 2, Nakul K. Chaudhary, Manish Bansal, Brihaspati Narayan Shukla, Saikat Boliar, Bimal K. Chakrabarti, Jayanta Bhattacharya HIV Vaccine Translational Research Laboratory, Translational Health Science & Technology Institute, Gurgaon, India, 2 National AIDS Research Institute, Pune, India Background: HIV- clade C has been widely shown to be resistant to neutralization by ggb2 monoclonal antibody (MAb) in different geographic settings. In the present study, using a IgGb2 sensitive and a resistant HIV- India clade C envelope (Env), we examined determinants modulating IgGb2 sensitivity. Methods: Chimeric envelopes were made by PCR and site-directed mutagenesis followed by blunt end ligation. The susceptibility of Envpseudotyped viruses to MAbs was assessed in a TZM-bl reporter cell based neutralization assay. Exposure of neutralizing antibody epitopes were assessed by virus-mab washout assay. Results: We identified determinants in V2/C2 region that governed susceptibility of Env-pseudotyped viruses to IgGb2, however no effect was found against soluble CD4 and anti-cd4 MAb. Interestingly, we found six discontinuous residues within the V2/C2 region in the IgGb2 resistant envelope that significantly modulated neutralization of Envpseudotyped viruses to PG9/PG6 MAbs. The reciprocal neutralization susceptibilities of Env-pseudotyped viruses to IgGb2 and PG9 MAbs by determinants in V2/C2 region were correlated with increased exposure of their corresponding epitopes. Conclusions: Our study highlighted vulnerabilities in V2/C2 region that alters the recognition of HIV- clade C primary viruses by IgGb2 and PG9 MAbs

230 Posters Posters 6: Passive Antibody Functions P6.07 Utilizing Mucin-tethered HIV IgG to Enhance HIV Vaccine Function Jeffrey R. Schneider, Maryam Shansab 2, Bronwyn Gunn 2, Shaunna Shen 3, Judith Lucas 3, Archer Smith, Neil Kelleher, Georgia Tomaras 3, Galit Alter 2, Ron Veazey 4, Thomas Hope Northwestern University, Chicago, IL, United States, 2 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 3 Duke Human Vaccine Institute, Durham, NC, United States, 4 Tulane National Primate Research Center, Covington, LA, United States P6.08 Antibody Inhibition of HIV- Transmission from Antigen-presenting Cells to CD4 T Lymphocytes Involves Immune Cell Activation Bin Su, Alexandre Lederle 2, Géraldine Laumond 3, Sylvie Schmidt 3, Thomas Decoville, Camille Ducloy, Christiane Moog INSERM U 09, FMTS, Université de Strasbourg, Strasbourg, France, 2 INSERM U 955 Université Paris-Est Créteil / The Vaccine Research Institute, Créteil, France, 3 INSERM U 0, FMTS, Université de Strasbourg, Strasbourg, France Background: Our lab has recently reported that antibodies can tightly bind mucus found in the female reproductive tract. We have identified several specific interactions between subsets of IgG and specific mucins with the best-defined example being HIV IgG and MUC6. Binding of IgG in HIV chronically infected individuals is increased 2-3 fold relative to the binding of IgG in healthy individuals. We are currently working to decipher which sub-population of HIV IgG is important for binding MUC6 and how these antibodies could be used to sequester incoming virions in the mucus of the FRT and enhance vaccine function. Methods: To purify human and rhesus IgG that associates with MUC6, we conjugated MUC6 to magnetic beads and performed a capture assay. A subset of IgG bound tightly to MUC6, requiring denaturation to remove them from bound MUC6. Therefore, we used 6M GuHCl to dissociate MUC6-specific antibodies from the magnetic beads. These antibodies were then interogated for their subclass and glycosylation content through mass spectrometry and antigen specificity through a Luminex assay. Results: HIV IgG binding to MUC6 is mediated through the FC portion of the antibody and there is an elevation of IgG subclass and fucose content. Macaque IgGs were isolated before and after SIV challenge and there was an elevation in MUC6 binding post-infection. Antibodies that bound tightly to MUC6 had elevated binding to gp4, but not gp20. Conclusions: Understanding the properties of HIV IgG antibodies that bind to mucus will aid our understanding of HIV infection. The observed enrichment of gp4 specific antibodies with MUC6, relative to gp20, suggest that the immune system has the ability to direct specific antigen responses to interact with a specific mucin. This observation reveals that it may be possible to direct vaccine-generated responses to accumulate in mucus. This would increase vaccine function by taking advantage of a new effector function that can facilitate virion trapping in mucus. Background: The mucosal tissues contain various HIV target cells including antigen-presenting cells (APCs) such as Langerhans cells (LCs), interstitial dendritic cells (idcs) or plasmacytoid (pdcs) in addition to CD4 T lymphocytes. These APCs play a major role in HIV- dissemination. Broadly neutralizing antibodies (bnabs) need to impair HIV- cell-tocell transmission to efficiently protect from HIV- transmission at the mucosal site. The aim of this study is to decipher the mechanisms by which Abs protect from HIV- transfer. Methods: We used a physiologically relevant model of primary APCs, infected with R5 HIV- isolates or transmitted/founder virus, cocultivated with autologous CD4 T cells in the presence or absence of bnabs. At 48 and 72h post-infection, the percentages of infected cells, the expression of intracellular SAMHD and CD83 + /CD86 + maturation markers were determined by flow cytometry. IFN-α production was measured in the culture supernatant. Results: We found that APCs efficiently transferred HIV- to adjacent CD4 T cells. Interestingly, coculture with CD4 T cells downregulated SAMHD expression in DCs, enhanced HIV- replication, induced DC maturation and increased IFN-α secretion. bnabs efficiently prevented HIV- transfer to CD4 T cells with a similar efficiency as cell-free infection. Moreover, DC maturation and IFN-α production were modulated by Abs. For example, bnab VRC0 did not impair DC maturation and IFN-α secretion, whereas 4E0 increased immune cell activation. This Abassociated modulation of APC activation participates to the overall Ab inhibition of HIV- transmission. Conclusions: During HIV cell-to-cell transmission, crosstalk between APCs and autologous CD4 T lymphocytes occurs leading to increased HIV replication, immune activation and immune sensing. HIV--specific Abs modulate these immune functions, therefore interfering with HIV- transmission. Consequently, this additional Ab function should be taken into consideration for the design of new vaccine strategies. 230 HIV Research for Prevention 204 HIV R4P

231 Posters 6: Passive Antibody Functions Wednesday, 29 October P6.09 Functional Screening of Human Broadly Neutralizing HIV- Monoclonal Antibodies by Antibody Cell Surface Display Zehua Sun, Jingjing Li, Zheng Yang, Yu Zhang, Shiqiang Lu, Meiyun Zhang AIDS Institute, Department of Microbiology, Faculty of Medicine, The University of Hong Kong, Hong Kong, Hong Kong Background: Identification of broadly neutralizing HIV- monoclonal antibodies (bnmabs) can facilitate the development of an effective HIV- /AIDS vaccine and aid for prevention and treatment of HIV- infection. A lot of bnmabs against HIV- were isolated in the past three years, most of which were isolated based on their binding activities to HIV- wild type or engineered envelope glycoprotein (Env). Methods: Here, we developed a novel methodology for isolating HIV- bnmabs based on antibody neutralization activity by displaying fulllength antibody libraries on target cell surface followed by sorting the cells by antibody neutralization ability. Results: After several rounds of sorting, a panel of human mabs has been isolated that can neutralize various isolates from different clades when displayed on the surface of mammalian cells. Several isolated antibodies have been converted into soluble version and characterized. Three mabs can neutralize several Chinese circulating tier 3 viruses. These antibodies exhibited neutralizing profiles that are complimentary with that of b2. We are currently characterizing the epitopes recognized by this panel of antibodies. Conclusions: Functional screening of HIV- bnmabs may help elucidate the mechanisms of antibody-mediated protection against viral infection. 23

232 Posters Posters 7: Post-exposure Prophylaxis P7.0 Safety and Tolerability of Co-formulated Tenofovir/Emtricitabine/Elvitegravir/ Cobicistat ( Quad ) for Non-occupational Postexposure Prophylaxis P7.02 HIV Seroconversions among Men who Have Sex with Men who Used Non-occupational Post-exposure Prophylaxis at a Boston Health center from Kenneth H. Mayer,2,3, Charles Gregor, Marcy Gelman, Chris Grasso, Kathy Melbourne 4, Matthew J. Mimiaga,5,6 Fenway Health, Massachusetts, Boston, MA, United States, 2 Beth Israel Deaconess Medical Center, Infectious Diseases/Medicine, Boston, MA, United States, 3 Harvard Medical School, Medicine, Boston, MA, United States, 4 Gilead Sciences, Clinical Research and Education, Foster City, MA, United States, 5 Harvard School of Public Health, Epidemiology, Boston, MA, United States, 6 Massachusetts General Hospital, Psychiatry, Boston, MA, United States Background: Post-exposure prophylaxis (PEP) to prevent HIV transmission after high risk exposures has been recommended for more than a decade, but older regimens using 3 medications have been inconvenient and/or had many side effects. Quad contains one integrase, and 2 reverse transcriptase, inhibitors, and could be used as a single pill, taken once a day for PEP. Methods: Since 997, a Boston community health center has provided NPEP. Between 5/3 and 3/4, 48 patients (pts) who presented for PEP agreed to initiate a Quad regimen. Analyses include descriptive findings, chi-square and Fisher exact tests to compare symptoms and completion rates with Quad NPEP compared to prior PEP regimens used at this site. Results: Of 48 enrolled pts, 47 were MSM and one was a woman. Most (72.9%) were White, 0.4% Black; their ages ranged from 20 to 62 y.o. The study retention rate was 95%. One pt was switched to another regimen when his baseline creatinine was elevated. Another pt discontinued Quad after reporting acute side effects, but later disclosed concomitant use of TDF/FTC/Efavirenz. One pt discontinued Quad because of diarrhea, which was noted by 44%, but usually (88%) was mild, and often self-limited. Other common adverse events were: nausea/ vomiting (3%), fatigue (25%), flatulence (25%), abdominal discomfort (5%), and abnormal dreams (3%). These symptoms were usually mild, and there were no severe adverse events reported. In comparison, prior Fenway PEP pts who used 3 drug regimens containing protease inhibitors more often reported nausea/vomiting, fatigue, but less often reported abdominal discomfort (all p s< 0.0). Pts who used TDF/FTC/ Ral for PEP less often reported diarrhea (p < 0.0), but more often missed doses than Quad pts (p=0.02). Overall self-reported adherence with Quad PEP was 99.%. No Quad pts became HIV-infected during the period of observation. Conclusions: Quad PEP seems safe and well-tolerated after risky sexual exposures, with a minimal pill burden and self-reported high completion rates. Sachin Jain,2,3, Catherine E. Oldenburg 4, Matthew J. Mimiaga 2,3,4, Kenneth H. Mayer,2,3 Beth Israel Deaconess Medical Center, Boston, MA, United States, 2 The Fenway Institute, Boston, MA, United States, 3 Harvard Medical School, Boston, MA, United States, 4 Harvard School of Public Health, Boston, MA, United States Background: Non-occupational post-exposure prophylaxis (NPEP) has been recommended for biomedical HIV prevention for nearly 20 years. However, limited behavioral and outcome data exist for men who have sex with men (MSM) who present for NPEP. Here, we report sociodemographic and behavioral characteristics of MSM NPEP users and factors associated with subsequent HIV infection at one center over 6 years. Methods: A retrospective longitudinal study of electronic medical records of NPEP users between July, 997 and August, 203 was performed at a large community health center in Boston. Eligible participants were age 8 years, HIV-uninfected, and had possible nonoccupational exposures to HIV. Cox proportional hazards models were used to assess factors associated with HIV seroconversion. Results: Data from 788 MSM were analyzed. Median age at first visit was 32.9 years. Thirty-nine patients became HIV-infected, 35 of which occurred >6 months post-exposure. Consensual unprotected sex (64.2%) was the leading reason for NPEP, including receptive anal (63.%), insertive anal (33.2%), and receptive oral (6.5%). The source partner s HIV serostatus was unknown for 64.4%, positive with unknown treatment status for 8.%, and positive and not on treatment for 4.%. Among consensual exposures, the proportion of presentations due to unprotected sex increased by year (RR=.0 per year;p=0.02). HIV incidence was 2.2 infections per 00 person-years. MSM with incident HIV tended to be younger (AHR=0.93;p=0.002), African American (AHR=3.59;p=0.04), and were more likely to be previously infected with Hepatitis B (AHR=.88;p=0.0). Repeat NPEP use, ranging from 2 to 5 courses, was not associated with HIV infection (AHR=0.67;p=0.26). Conclusions: NPEP users in this study demonstrated a high HIV incidence. Most of the HIV infections occurred >6 months post-exposure and likely represent recurrent high-risk exposures. Younger and African American MSM may benefit from early HIV risk-reduction and preexposure prophylaxis counseling. 232 HIV Research for Prevention 204 HIV R4P

233 Posters 8: Preclinical Evaluation of Vaginal Films and Gels Wednesday, 29 October P8.0 Characterization of a Dapivirine Impurity Found in Dapivirine Gel Product During Storage P8.02 A Hydrogel Tissue Model for Evaluation of Triple-antiretroviral Electrospun Fibers as a Microbicide Tiffany Derrick, Stephen Ampofo, Brid Devlin International Partnership for Microbicides, Product Development, Silver Spring, MD, United States Anna Blakney, Kim Woodrow University of Washington, Bioengineering, Seattle, WA, United States Background: Dapivirine (DPV) has been formulated as a gel drug product containing sorbic acid as a preservative for clinical trials. Upon stability of developmental DPV gel batches, impurities were detected in the drug product as well as the reaction vessels that contained both DPV and sorbic acid. This abstract summarizes the isolation and characterization of these impurities and preliminary assessment of their toxicological significance. Methods: Dapivirine Gel, 0.2%, samples were placed in storage chambers maintained at 60⁰C to generate the product impurities found in the clinical gel batch. The impurities were extracted by solvent-solvent extraction procedures, separated and purified through the use of two semi-preparative HPLC procedures in sequence. The compounds were analyzed by Mass Spectrometry and NMR. The major component was isolated in sufficient quantity to allow for advanced NMR investigation. Results: The mass spectra of the major product impurity gave a molecular ion at m/z of 429, which is consistent with its assignment as a DPV sorbic acid adduct. This was confirmed by the H NMR spectra which showed signals attributable to DPV and a multiplicity pattern as those for sorbic acid. At elevated temperatures and when dissolved in deuterated DMSO, the compound converted to DPV (m/z 329), evidenced by NMR and MS data. Conclusions: The observed behavior of the impurity under different experimental conditions suggests that under thermal stress the impurity sheds the sorbic acid to retain the DPV only component of the complex. Thus, while the proposed structures for the impurities could not be definitively confirmed, the body of evidence accumulated from the study supports the assignment of DPV sorbic acid adducts to the impurities. The complex is loosely associated therefore the adduct formation is most likely reversible. Hence, the toxicological profile of these impurities should be similar to that of the parent compound, which has no reported safety concerns in human. Background: Currently, there are no pre-clinical models that allow for rapid and high-throughput evaluation of topical microbicide dosage forms that enable correlation of drug concentration in tissue with material properties. Poly-hydroxyethyl methacrylate (phema) hydrogels have been widely used in tissue engineering, and can mimic vaginal tissue. phema hydrogels were used to evaluate electrospun fibers, which have recently been developed as a novel microbicide dosage form for vaginal delivery of antiretrovirals (ARVs) but have yet to be evaluated in animal models. Fibers containing three ARVs with varying thickness, drug loading and stacking order were evaluated for release into the phema tissue mimic. Release of drug from electrospun fibers into the hydrogels was then compared to release into polarized, human vaginal tissue. Methods: phema hydrogels were prepared by UV-polymerizing a mixture of 58.8% (v/v) hydroxyethyl methacrylate, 0.6% (v/v) ethylene glycol dimethacrylate, and 0.6% benzoin isobutyl ether and 40% (v/v) water. A 0% (w/v) solution of polyvinyl alcohol in water was electrospun using a production-scale instrument (NS WS500U, Elmarco, Inc.). Fibers were loaded with dapivirine (DPV), maraviroc (MVC), and/or tenofovir (TFV). Drug concentration in the tissue mimic and vaginal tissue was analyzed using HPLC and LC-MS/MS, respectively. Results: phema hydrogels contain ~70% (w/w) water, which is similar to that of vaginal tissue. Drug extraction from phema hydrogels was validated using known amounts of DPV, MVC and TFV and >90% recovery of each drug was achieved. Drug release from electrospun fibers was found to be slower with increasing fiber thickness, combination drug loading and orientation of hydrophobic layer adjacent to the tissue mimic. Drug release was also impacted by free fluid volume and composition. Conclusions: Based on these results, the lead fiber-based microbicide design will be further evaluated using animal models. phema is an adequate tissue model for development of microbicides

234 Posters Posters 8: Preclinical Evaluation of Vaginal Films and Gels P8.03 Use of the ICCA to Predict Dosing of HIV Microbicide Products to Achieve Virus Sterilization in Target Tissue Karen W. Buckheit, Caitlin A. Buchholz, Cory Shetler 2, Charlene S. Dezzutti 2, Pedro M. Mesquita 3, Betsy C. Herold 3, Robert W. Buckheit, Jr. ImQuest BioSciences Inc., Frederick, MD, United States, 2 University of Pittsburgh Medical Center and Magee-Women Research Institute, Pittsburgh, PA, United States, 3 Albert Einstein College of Medicine, Bronx, NY, United States P8.04 Toxicological Analysis of Avaren-Fc: A Potential HIV Microbicide Tiffany N. Grooms-Williams, Joseph Kouokam 2, Alfred Jenson, Nobuyuki Matoba 3,4 University of Louisville School of Medicine, James Graham Brown Cancer Center, Louisville, KY, United States, 2 University of Louisville, Owensboro Cancer Research Program, Louisville, KY, United States, 3 University of Louisville School of Medicine, Department of Pharmacology and Toxicology, Louisville, KY, United States, 4 Owensboro Cancer Research Program, Owensboro, KY, United States Background: The historical dosing strategy employed for HIV prevention products has been to flood target tissue with high nontoxic concentrations of a product. Continuation of this strategy results from the lack of data from a successful clinical product or dose response studies in a predictive animal model with direct correlation to use in humans. We have hypothesized that sensitive in vitro assays can be used to define the required tissue concentration of a microbicide to totally prevent HIV transmission and/or spread of virus from initially infected sites. Methods: The microbicide transmission and sterilization assay (MTSA) has defined sterilizing concentrations of microbicide agents. For example 00 nm of the NNRTI IQP-0528 and 7 µm of the NtRTI tenofovir are required to sterilize a virus infected cell culture. Through continued evolution of the MTSA and efforts to better understand the biology of sterilization, we have developed an infectious cell center assay (ICCA) to more sensitively evaluate sterilization by microbicides under conditions in which these products must act. For IQP-0528 the ICCA yielded identical sterilizing concentrations as the MTSA (00 nm) and correlated the sterilizing concentrations with effective concentrations observed in antiviral and pharmacodynamic assays employing cervical explant tissue. Results: Protection of mucosal tissue from HIV challenge required approximately 0 µm of formulated IQP-0528 (00 µm with unformulated IQP-0528). Performance of antiviral assays with lysed explant tissue that had been soaked with 0 µm IQP-0528 also yielded complete protection of target cells, indicating that the 0 µm dose delivered a sterilizing concentration of IQP-0528 to the tissue. Conclusions: Our efforts now involve understanding the tissue pharmacokinetics of microbicides in order to better understand not only the concentration of product but the timing of dosing relative to the time of infection and rate of product uptake. Background: HIV continues to be a serious threat to the global public health, especially among women in developing countries. Microbicides may offer an alternative approach for blocking transmission of HIV when condom use cannot be negotiated with male partners. We have developed a novel HIV microbicide candidate. Avaren-Fc (AvFc), consisting of oligomannose-specific lectin Avaren dimerized by fusion to the Fc region of human IgG, has shown low-nanomolar antiviral activity against multiple HIV type and type 2 clinical isolates. Moreover, AvFc induced antibody-dependent cell-mediated cytotoxicity. Cognizant of the imperative that microbicides not induce epithelial damage or inflammatory responses, here we show that AvFc is nonirritating and noninflammatory in human peripheral blood mononuclear cells (hpbmcs), in human vaginal epithelial tissue (hvet), and in vivo in the rabbit vaginal irritation (RVI) model. Methods: Human PBMCs were treated with AvFc up to 00µg/ml, nonoxynol-9 (N9), or Concanavalin A (ConA) and accessed for viability, induction of activated cells, and induction of proinflammatory cytokines and chemokines. Reconstituted hvet grown on membrane filters were exposed to AvFc up to 0.% (w/v), formulation buffer, or 0.2% N9 for 72 hours. Using the RVI model, rabbits were exposed daily to AvFc up to 0.% (w/v), vehicle control, or 2% Benzalkonium chloride (BZK) for 0 days. In both models, tissue damage was evaluated by histological assessment. Furthermore, viability was evaluated by MTT analysis, and barrier disruption was accessed by transepithelial electrical resistance in the hvet. Results: Unlike ConA or N9, AvFc did not show any cytotoxicity, proinflammatory cytokine release or mitogenicity in hpbmcs at maximal antiviral concentrations. In addition AvFc did not induce any discernible toxic or inflammatory effects in both vaginal models compared to N9 or BZK. Conclusions: Taken together, these results highlight AvFc s candidacy as a safe microbicide with broad and potent anti-hiv potential. 234 HIV Research for Prevention 204 HIV R4P

235 Posters 8: Preclinical Evaluation of Vaginal Films and Gels Wednesday, 29 October P8.05 Comparative Pharmacokinetics of DS003, a gp20 Binder and Candidate Microbicide in Rats, Rabbits and Dogs Jonathon Holt, Adam Leggett 2, Leon Beaudoin 2, Jeremy Holding 2, Whenhao Zhu 2, Annabell Pitcher 2, Jeremy Nuttall IPM, Silver Spring, MD, United States, 2 Huntingdon Life Sciences, Huntingdon, United Kingdom P8.06 Dapivirine and Tenofovir Vaginal Films and Gels: Macaque Pharmacokinetics Dorothy L. Patton, Yvonne Sweeney, Lisa C. Rohan 2,3, Mark Marzinke 4, Craig W. Hendrix 4, Sharon Hillier 3 University of Washington, Obstetrics & Gynecology, Seattle, WA, United States, 2 University of Pittsburgh, School of Pharmacy, Pittsburgh, PA, United States, 3 Magee Women s Research Institute, Pittsburgh, PA, United States, 4 Johns Hopkins University School of Medicine, Clinical Pharmacology, Baltimore, MD, United States Background: DS003 is a novel gp20 binder selected for development as a microbicide due to its potent and broad anti-hiv- activity, and its low bioavailability. Systemic exposure was determined during vaginal and oral toxicity studies in rats, rabbits and dogs, and plasma protein binding and metabolism in hepatocytes were evaluated in vitro. Methods: Female rats, rabbits and dogs received 000mg/kg DS003 orally in 2% methylcellulose (rat and rabbit) or a gelatin capsule (dog). A DS003 gel (up to 25% w/w) was dosed vaginally using a fixed dose volume for rats (0.2mL) and rabbits (ml), and a body weight-adjusted dose volume for dogs (ml/kg). Plasma samples were collected for toxicokinetic analysis. In vitro, plasma protein binding was evaluated in rat, rabbit, dog and human by equilibrium dialysis, and the rate and extent of metabolism was determined in hepatocytes from rat, dog, rabbit, monkey and human. Detection of DS003 and metabolites was by LC-MS/MS. Results: Systemic exposure was low via both dosing routes with the max. mean plasma conc. being.9 µg/ml after a 000mg/ kg oral dose in rats. Relative to dose, exposure in all species was consistently higher after vaginal than after oral dosing. Dose-adjusted mean C max and AUC 24 were in the order rat/vaginal>rat/oral>dog/ vaginal>rabbit/vaginal>dog/oral>rabbit/oral. The orders for the rate and extent of metabolism were rabbit>dog>rat>monkey>human, and rabbit>monkey>human>dog>rat, respectively, suggesting that metabolism may account for the differences in exposure between species. However, first pass metabolism was unlikely to account for the difference between vaginal and oral dosing. Plasma protein binding was generally >90%. Conclusions: Systemic exposure to DS003 was low, particularly after oral dosing, and differences observed between species may be due to variations in metabolism. The low bioavailability of DS003 supports its development as a microbicide, but presents challenges for nonclinical assessment of systemic toxicity and safety pharmacology. Background: Pharmacokinetics (PK) of 5 microbicide formulations were assessed in the macaque model. Formulations reduced in size and dosage for macaque use were: film and gel delivering dapivirine (DPV, 0.5mg, 0.75mg respectively); 2 films and gel delivering tenofovir (TFV, 3.4mg,.2mg, 5mg). Methods: For each formulation, 8 macaques received one vaginal dose followed by blood and tissue samples for PK analysis. Plasma was collected prior to, and hr, 2hr, 4hr, 6hr, 8hr, day (d), 2d, and 7d post exposure. Cervical and vaginal biopsies were collected at 6hr, d, and 7d after DPV exposure; vaginal biopsies were collected d and 7d after TFV exposure. Results: Plasma DPV levels peaked at median (IQR) 240 (76, 304) pg/ ml after film compared to 23 (59, 25) pg/ml after gel administration. Dose-adjusted peak plasma levels trended toward being higher (p=0.065) with the film. Overall dose-adjusted drug exposure (areaunder-the-curve [AUC]) was not different between DPV formulations. Higher vaginal tissue levels of DPV were measured after gel compared to film dosing (median 223pg/sample vs 09pg/sample), at 6hr (p=0.029). DPV concentrations were higher in vaginal vs cervical tissues for both formulations at 6hr (p=0.023). All tissue levels were < pg/ sample 7d after drug exposure. Plasma TFV levels peaked median 4hr after film vs hr after gel dosing; dose-adjusted values were not different for Cmax or AUC among all 3 formulations (NS). Dose-adjusted tissue concentrations of TFV and TFV- DP were generally higher with film compared to gel (p< 0.038) at d (highest) and 7d after dosing. All tissue levels were far below peak, but still detectable 7d after dosing in at least half of those receiving film and 3 of 8 receiving gel. Conclusions: Gel delivered more DPV to tissues than film with similar plasma exposure. TFV plasma levels were reflective of the dose delivered, regardless of platform. Film formulations delivered more TFV and TFV-DP to tissues than gel

236 Posters Posters 8: Preclinical Evaluation of Vaginal Films and Gels P8.07 Novel Intravaginal Drug Delivery Systems of the Antiviral for Prophylaxis of HIV/AIDS and Prevention of Sexually Transmitted Diseases Mahesh R. Sherkar, Ashok V. Bhosale 2, Mala D. Menon 3 Shri Vivekanand Nursing Home Trust s College of B Pharmacy, Pharmaceutics, Rahuri, India, 2 Pune District Education Association s Seth Govind Raghunath Sable College of Pharmacy, Pharmaceutics, Saswad, Pune, India, 3 Bombay College of Pharmacy, Kalina, Santacruze(E), Pharmaceutics, Mumbai, India P8.08 Alternative Tenofovir Gel Formulations: Safety and Drug Absorption in the Macaque Model Yvonne Sweeney, Dorothy L. Patton, Lisa C. Rohan 2,3 University of Washington, Obstetrics & Gynecology, Seattle, WA, United States, 2 University of Pittsburgh, School of Pharmacy, Pittsburgh, PA, United States, 3 Magee Women s Research Institute, Pittsburgh, PA, United States Background: Male to female transmission is eight times more likely to occur than female to male transmission due to the anatomy of the vagina, socio-economic factors, and the disempowerment of women, unable to refuse unsafe sexual practices in some communities. Currently, the only prevention strategy available by using condoms or abstaining from sexual intercourse, but 67% did not use condoms consistently and 3% had never used. The increased incidence of HIV in women has identified the urgent need for efficacious and safe intravaginal delivery of antiviral that can be used and controlled by women. Methods: Vaginal gels were prepared by ionic gelation process. Water soluble antiviral abacavir/ didanosine was loaded in a hydrophilic polymer chitosan (CS) and a diblock copolymer of (PEO-PPO), cross linked with tripolyphosphate (TPP), to form nanoparticals were evaluated and added into slurry of polymeric solution of carbopol/hpmc to form a gel, and evaluated. Results: Nanoparticals size ranges nm, DSC was c, had Zeta potential 40 mv, FTIR peak found at 664cm -, loading efficacy 74%. Gel was uniform, transparent had ph 7.4. Gelling capacity and extrudability was good, viscosity 30000Cp, Drug content78%, spreadability 72.87gm.cm/sec. Irritation, reddening not showed in vaginal irritation study. In-vitro drug release for nanoparticals was 70%, and for gel was up to 55%. Accelerated stability study for 2 weeks at 37 0 c/45 0 c/60 0 c nanoparticals and gel were stable and potent. Conclusions: This research has led to the design, development and evaluation of novel antiviral formulations to be employed for the prophylaxis of HIV/AIDS and prevention of STD used by women. Polymers used are safe, biocompatible, mucoadhesive and having good antiviral entrapment capacity produces local effect on the vaginal mucosa, and controlled release for the antiviral with minimal side effects. Half life of the water soluble antiviral will be increased ultimately decrease the dose and dosing frequency, and cost of therapy. Background: Two reformulated gels designed to deliver tenofovir (TFV) in a mucosa friendly manner for use as dual compartment microbicides were assessed in the pigtail macaque model. A reduced glycerin (RGVF), an iso-osmolar (TFIO), and matched placebo gels were assessed for safety and drug absorption with vaginal use. Methods: Six macaques completed each study arm, receiving one of the TFV or placebo gels, daily for four days. Safety of repeated, daily exposures was measured by repeated colposcopic assessment, vaginal ph, vaginal smear and microbiology tracking. In addition, vaginal swabs and blood samples were collected daily for TFV detection. Rectal swabs were also collected to determine whether TFV crossed from the site of delivery to the rectum. Finally, vaginal and cervical biopsies were collected on study day 5 to document TFV and its active metabolite tenofovir diphosphate (TFV-DP) levels in the local tissue. Results: RGVF and TFIO caused no discernible irritation to cervicovaginal tissues in the standardized safety study. RGVF was tolerated slightly better than its placebo, while TFIO was indistinguishable from its placebo gel by parameters assessed here. Both TFV gels resulted in heightened plasma TFV levels (RGVF 25ng/mL; TFIO 40ng/mL) 30-minutes after the initial exposure (T30 only measured on day ), and in much lower concentration (-2ng/mL) through day 5. TFV accumulated in the vaginal secretions, peaking on day 5 for both gels (RGVF 39,372 (57-38,000)ng/sample; TFIO 28,808 ( ,750)ng/sample), and dropping precipitously (< 60ng/sample) on day 8, four days after the last gel application. Rectal secretions tested positive for TFV at greatly reduced levels. TFV and TFV-DP were detected in cervical and vaginal biopsies; TVF-DP was available in higher concentrations in the vaginal tissues compared to cervix. Conclusions: Both of these alternate formulations of tenofovir gel are safe to the mucosal environment and efficacious in delivering tenofovir drug, with vaginal use. 236 HIV Research for Prevention 204 HIV R4P

237 Posters 9: PrEP Implementation P9.0 Wednesday, 29 October P9.02 HIV Negative Partners in Sero-discordant Relationships Are a forgotten Lot: Room for PrEP Maureen Akolo, Joshua Kimani, Larry Gelmon University of Nairobi/Manitoba, Nairobi, Kenya Background: University of Nairobi/ University of Manitoba HIV/STI research team has been managing a mother to child HIV prevention clinic (MCH) targeting low risk population in Pumwani since 988. In 2006, it opened up to all HIV infected individuals to improve access to the much needed ARVs services. Prevention with Positives (PwP) initiative was rolled out in Where service providers encourage index clients enrolled to disclose their status to sex partners and bring them for HIV testing. The efforts lead to the establishment of the serodiscordant couples cohort. Methods: A longitudinal descriptive study has been carried out among sero-discordant couples since September 2008 up to December 203. Questionnaires were used to collect baseline and resurvey information on demographics, condom use, risk taking behaviors etc from partner of the index clients. HIV rapid test are conducted every six months and their PrEP knowledge ascertained. Results: Out of 420 partners interviewed, baseline data showed; only 6% were using condoms correctly and consistently while 37% did not use them at all. 63 % reported having other sex partners of which 23% reported more than 50 life time sex partners. Off the latter group, 45 % reported to be married elsewhere but kept the index patient as a regular sex partner. At baseline % had knowledge of PrEP but only 2% were willing to use PrEP. However through behavioral change communication 60% are willing to take up PrEP with 88% reporting condom use. In the past year, 8% of these partners have been treated for an STI, no couples agreed to initiate treatment as prevention (TASP). Luckily, only 2% HIV sero conversions noted. Conclusions: HIV negative individuals in discordant relationships should be labeled key populations. They are highly sexually active with multiple sex partners. Access to PrEP should be mandatory to support ongoing HIV prevention strategies. Integrated Next Step Counseling for Sexual Health Promotion and Medication Adherence for Individuals Using Pre-exposure Prophylaxis Christopher Chianese, K. Rivet Amico 2,3, Kenneth Mayer, Albert Liu 4,5, Vanessa Mcmahan 6, Sybil Hosek 7, Megha Mehrotra 6, Robert Grant 4,6 Fenway Health, The Fenway Institute, Boston, MA, United States, 2 Center for Health, Intervention, and Prevention, University of Connecticut, Storrs, CT, United States, 3 Applied Health Research, Brighton, MI, United States, 4 University of California, San Francisco, CA, United States, 5 San Francisco Department of Public Health, San Francisco, CA, United States, 6 Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA, United States, 7 Troger Hospital of Cook County, Department of Psychiatry, Chicago, IL, United States Background: In the iprex Open Label Extension (OLE) study counselors integrated Next Step Counseling (NSC- developed by iprex to promote adherence) with sexual health promotion counseling to produce Integrated Next Step Counseling (insc). This approach combines sexual health promotion and PrEP-adherence counseling into a single discussion. We evaluated insc session records from US iprex OLE study sites (Boston, Chicago, San Francisco) to identify main themes of discussions relevant to sexual-health promotion practices and PrEP adherence. Methods: insc entails a brief discussion between counselors and participants based on demonstrated health behavior models and motivational communication strategies. The approach includes a two-part discussion that explores contextual facilitators, barriers and individualized needs for enhancing sexual health promotion (part ) and repeats this kind of discussion specific to PrEP adherence (part 2). Counselors documented facilitators, barriers, and needs on a record form at the end of each insc session. Results: insc was administered to 287 participants (regardless whether on or off PrEP) between June,20 and September,203 producing a total of 2492 sessions for review with 576 sessions also including PrEPadherence for 223 on-prep participants. The most reported facilitator of safer sex practices included personal commitment (motivation) to staying HIV-negative (49%). Getting caught up in the moment (2%) was the most common identified barrier. For insc adherence discussions, matching dose taking with a routine daily event (86%) was the most common facilitator; disruption in routine the most common challenge (37%). Challenges and needs were not identified for 42% and 49% of sessions, respectively. Conclusions: The insc discussions identified factors participants found relevant in promoting their sexual-health practices and PrEP adherence. Future explorations into other factors that are influential to the promotion of sexual-health practices and PrEP adherence are needed

238 Posters Posters 9: PrEP Implementation P9.03 P9.04 Community-driven Strategies for the Use of ARV-based Prevention: Outcomes from U.S. City Workshops Jessica Terlikowski, Molly Morgan Jones 2, Jim Pickett, Gavin Cochrane 2, Jennie Corbett 2, Joanna Chataway 2 AIDS Foundation of Chicago, Chicago, IL, United States, 2 RAND Europe, Cambridge, United Kingdom Background: Complex challenges accompany implementation of ARVbased HIV prevention, including PrEP, treatment, microbicides and PEP. Social, political, economic and technological factors must be considered - scientific data alone is not sufficient. Mapping Pathways partners AIDS Foundation of Chicago and RAND Europe address the U.S. community s role in research and implementation. Methods: Three two day scenario development workshops were conducted in San Francisco, Atlanta and Washington, D.C. in 203. Stakeholders, including researchers, public health staff, prevention programmers, policy experts and advocates imagined the year 2025 and forecasted the potential outcomes of ARV-based implementation strategies. Workshop participants built optimistic, pessimistic, and mixed outcome scenarios using the social, political, economic and technological factors they identified. Mapping Pathways analyzed the issues and synthesized the scenario outcomes across workshops to create an integrated future scenario as a pathway forward. Results: An integrated approach to HIV prevention is needed. Advances in biomedical and digital health technologies must be tied to the social contexts in which they will be implemented. Changes in healthcare systems, such as the advent of the Affordable Care Act, offer opportunities and challenges to the current infrastructure of HIV care and prevention service delivery. It is imperative to capitalize on the HIV community s track record of developing innovative programs and systems to provide holistic care. Conclusions: The dynamic scientific evidence base must be synthesized and integrated into kinetic political, social, economic, and technological factors that vary by geography, population, and infrastructure. This ever-shifting environment provides opportunities and challenges for the implementation of new ARV-based HIV prevention strategies, as the future is driven by communities. Gay Men Involved in HIV: Their Concerns and Hopes about PrEP Marc-André LeBlanc Consultant, Gatineau, QC, Canada Background: There has been much debate about pre-exposure prophylaxis (PrEP). Gay men involved in HIV-related work are more likely than many other stakeholders to know about PrEP and to have formed opinions about it. Methods: In June 203, 60 HIV-positive and HIV-negative gay men involved in HIV-related work were asked to complete a brief, informal, anonymous online survey on their opinions about PrEP. 7 HIV-negative men and 3 HIV-positive men responded to the 3 open-ended questions: ) Can you ever imagine a time your life past, present or future when you might have considered using PrEP if it was available? Why? Why not? 2) Given the situation around HIV in your community today, what is your biggest concern about PrEP? 3) Given the situation around HIV in your community today, what is your biggest hope for PrEP? Results: In response to the question about using PrEP themselves, HIV+ gay men were evenly split: half thought they would have used PrEP if it had been available when they were negative, and half thought they would not have used it. Most of the men who would not have used it mentioned lack of consciousness about their own HIV risk and lack of awareness of HIV prevention as the main reasons. In contrast, the vast majority of HIV- men could imagine a time when they would consider using PrEP. Both HIV+ and HIV- men were generally in agreement about their hopes and concerns about PrEP. Likewise, similar themes emerged for both hopes and concerns, including: access, prioritizing and reinvigorating prevention efforts, identifying good PrEP candidates and supporting them, risk compensation, stigma, drug resistance, side effects, efficacy levels, medicalization of gay men, and desire for certain types of sex. Conclusions: Many gay men involved in HIV-related work can see themselves as potential PrEP users. Based on their experience, lack of self-awareness about risk and prevention options may be a significant barrier. Both HIV+ and HIV- gay men express remarkably similar hopes and concerns about PrEP. 238 HIV Research for Prevention 204 HIV R4P

239 Posters 9: PrEP Implementation P9.05 Wednesday, 29 October P9.06 Community Engagement and Male Involvement to Enhance Ring Adherence for Female Participants on a Phase III Microbicide Ring Trial Cheryl Emily Louw, Ntswaki Rose Masilo, Marthie de Villiers, Alinah Masiu, Annalene M. Nel 2, Michelle Isaacs 2, Smanga Ntshele 2, Colleen Herman 2 Madibeng Centre for Research, Brits, South Africa, 2 International Partnership for Microbicides, Paarl, South Africa Background: When recruiting female participants for a microbicide vaginal ring trial it is important to note that they are not only individuals, but also members of a community - women who are part of complex relationships and interactions. At Madibeng Centre for Research (MCR) it was noted that participants raised various concerns during their counselling sessions, which differed from partner-related issues to issues related to community members, who may have caused anxiety or discouraged ring use. Methods: With participants assistance, research nurses and the MCR investigators explored during counselling sessions concerns raised that could lead to possible trial non-compliance. Concerns identified were addressed in adherence counselling sessions to empower participants to comply with the protocol. These sessions targeted trial awareness, general knowledge about HIV and HIV risk to address personal risk awareness and were done on a one-to-one basis, during community events or, if consent was given, directly with male partners. Additional efforts were required for targeted males to intensify the support and assistance given to particular participants e.g. in cases where a partner did not understand the purpose of the trial or had safety concerns around the use of the vaginal microbicide ring. Results: With additional knowledge, the participants are more open to share their experiences with the community, male partners and also the site staff. It has been observed that the community and male partners support the work done by the study and the trial participants. Conclusions: Community awareness and male involvement are important to support women who are participating in a trial and encourage them to adhere to ring use, but community engagement efforts begin with the participants. Ready, Set, PrEP! A Multi-faceted Approach to Enhancing PrEP Knowledge among the HIV Workforce and Communities at Risk Jim Pickett, Jessica Terlikowski AIDS Foundation of Chicago, Chicago, IL, United States Background: The HIV workforce and at-risk communities need accurate, current information on PrEP and issues related to implementation and access. Not unlike the slut shaming surrounding contraceptives in the U.S., there is a fair amount of stigma and negative biases associated with PrEP, among service providers and communities who would benefit most from new prevention options. Project Ready, Set, PrEP! provides multi-faceted PrEP awareness and education activities in Chicago. Methods: Three activities work together to enhance education and awareness of PrEP in Chicago. ) trainings conducted by researchers, doctors, advocates, and PrEP users for the HIV prevention and care workforce; 2) talk show style community forums featuring celebrity hosts, entertainment, and open discussion of how PrEP fits into a holistic HIV prevention approach for gay men; and 3) the My PrEP Experience blog and Facebook page featuring PrEP users personal stories and informational resources. Results: Training participants pre- and post-tests revealed increases in PrEP knowledge, openness to PrEP, and comfort in talking with clients about PrEP. Community forum attendees reported through event evaluations, social media posts, and individual conversations that the events increased understanding of PrEP, reduced skepticism, and increased willingness to share their learnings with friends. The blog and Facebook page resulted in extensive dissemination of, and discussion about, PrEP educational information and the real-life experiences of PrEP users. Conclusions: Stigma and misinformation pose significant, but not insurmountable challenges to PrEP implementation. For PrEP implementation and emerging options to be successful, awareness must be built through tailored, dynamic strategies among people at elevated HIV risk and providers. Efforts in Chicago are contributing to better informed, less reactionary discourse about PrEP

240 Posters Posters 9: PrEP Implementation P9.07 LB Knowledge, Willingness and Intention to Use Pre-exposure Prophylaxis - PrEP - in France (204). Preliminary Results from a Community-based Survey Emmanuel Trenado, Daniela P. Rojas Castro,2, Guillemette Quatremère, Thomas Gonçalves, Jean-Marie Le Gall AIDES, Pantin, France, 2 Groupe de Recherche en Psychologie Sociale (GRePS), Lyon, France Background: In July 202, the FDA approved the daily use of TDF/FTC as PrEP for individuals at high risk for HIV infection. In May 204, the USA Public Health Service published the first clinical practice guidelines for PrEP and in July 204, WHO issued the consolidated guidelines on HIV prevention recommending use of PrEP as well. In France, the major HIV/AIDS NGO (AIDES) has been lobbying for temporary use of PrEP without success. In this context, AIDES conducted a survey aimed to know who would be interested in PrEP. Methods: A questionnaire exploring knowledge about PrEP, willingness and intention to use PrEP was proposed to all the people catered at AIDES from 3 st March-3 th April 204. A brief introduction to the questionnaire summarized what is PrEP. Results: 928 individuals answered the questionnaire. 76.2% of the respondents were French, 72.2% male and 42.4% had a university degree. Median age was % declared to be heterosexual men, 27.3% heterosexual or lesbian women, 34.3% MSM. Knowledge of PrEP before filling the questionnaire and intention to use PrEP were reported respectively by 45.5% and 89.5% of the respondents. Univariate logistic regression showed that factors associated with the intention to use PrEP are: being born abroad (p< 0,00), not having educational background (p< 0,00), having children (p< 0,05), being older (p< 0,05), having a HIV+ main partner (p< 0,0), already knowing the existence of PrEP (p< 0,05) and of the FDA approval (p< 0.05), willing to pay 20 Euros/month (P< 0,0), having >2 HIV tests in the last year (p< 0,05). Finally, self-perception of HIV acquisition risk as low was inversely associated with the intention to use PrEP (p< 0.05). Conclusions: These first results regarding knowledge, willingness and intention to use PrEP in France show that heterosexual migrants declare a higher intention to use PrEP. These results should be used to design an adequate PrEP delivery service. Further analysis will be presented in order to complete these results. 240 HIV Research for Prevention 204 HIV R4P

241 Posters 20: PrEP: Resistance, Modeling and Acceptability Wednesday, 29 October P20.0 How Much Do we Know about Drug Resistance due to PrEP Use? Analysis of Experts Opinion and Its Influence on the Projected Public Health Impact P20.02 Mathematical Modelling of the Impact of PrEP for Female Sex Workers and Men Who Have Sex with Men upon HIV Incidence and Survival in Southern India Dobromir Dimitrov, Marie-Claude Boily 2, Timothy B. Hallett 2 Fred Hutchinson Cancer Research Center, Statistical Center for HIV/AIDS Research and Prevention (SCHARP), Seattle, WA, United States, 2 Imperial College London, Department of Infectious Disease Epidemiology, London, United Kingdom Background: After four randomized clinical trials showing efficacy in different population, Truvada was approved for PrEP use. Public-health authorities need to carefully consider the plausibility of increasing spread of drug resistance following mass PrEP use. We investigate the current knowledge on drug resistance due to PrEP and its implications for decision making. Methods: We surveyed a panel of expert virologists to evaluate epidemiological relevance of key biological assumptions regarding drugresistance due to PrEP use. We identified major points of disagreement and studied how different beliefs about resistance propagate into the PrEP impact projected by mathematical models. We assumed the use of 90% efficacious PrEP by 50% of the population under epidemic conditions representative for South Africa. Results: Virologists disagreed on how fast resistance emerges in infected PrEP users (20-80 days) and both, positive or negative, correlations with adherence were thought possible. They differ on what level of protection PrEP retains against resistant HIV (25%-90%) and on the likelihood to transmit resistance (0%-75%). Based on the virologists opinion, we project % to 8% of the infected individuals with detectable resistance 0 years after the introduction of PrEP and 5% to 35% of the infected individuals at risk to fail ART as a result of past PrEP use. The rate of resistance emergence in infected PrEP users and the rate of reversion back to wild type after PrEP is interrupted are most influential on these resistance projections with larger levels of resistance outcomes associated with imperfect adherence. Conclusions: Infected individuals who develop resistance on PrEP may have no detectable levels of resistance after they interrupt PrEP use, but could be at risk to fail ART when initiated. Thus, the levels of detectable resistance due to PrEP, frequently used in modelling studies, may underestimate the potential resistance problem when PrEP is implemented in regions with overlapping ART regimens. Kate M. Mitchell, Holly J. Prudden, B M. Ramesh 2,3, Reynold Washington 2,4, Shajy Isac 2, S Rajaram 5, Fern Terris-Prestholt, Peter Vickerman,6 London School of Hygiene and Tropical Medicine, Global Health and Development, London, United Kingdom, 2 Karnataka Health Promotion Trust, Bangalore, India, 3 University of Manitoba, Winnipeg, MB, Canada, 4 St John s Research Institute, Bangalore, India, 5 CHARME-India Project, Bangalore, India, 6 University of Bristol, Bristol, United Kingdom Background: In Bangalore, HIV infection is concentrated amongst men who have sex with men (MSM), female sex workers (FSWs) and their commercial clients. Intervention programmes have succeeded in increasing condom use amongst these populations. We estimated the additional impact of targeted pre-exposure prophylaxis (PrEP) for highrisk MSM (HR-MSM) and/or FSWs. Methods: A deterministic model of HIV transmission between MSM, FSWs, commercial clients of FSWs and low-risk members of the general population was parameterised using data from Bangalore. The model was fitted to HIV prevalence data for MSM, FSWs and clients, and to ART coverage data for Bangalore from multiple time points, and was used to project the impact of PrEP targeted to FSW, HR-MSM or both groups, for a range of different PrEP coverage and efficacy assumptions. Impact was estimated in terms of infections averted (IA) and life-years gained (LYG), and efficiency was estimated in terms of years of PrEP per LYG. Results: Over 0 years, the model predicted that a PrEP intervention with 40% coverage of the target population and 60% efficacy could avert over a fifth of new infections amongst the targeted population (FSW or HR-MSM), and 2-3% of new infections in the whole population. Impact increased with higher efficacy and coverage. Targeting FSW had a greater population-level impact than targeting HR-MSM, and the effects of targeting both groups were almost additive. PrEP efficiency was predicted to be low but improved over time, with a median of 475 and 33 years of PrEP required per LYG after 0 and 20 years, respectively, when PrEP with 60% efficacy was used by 40% of HIV negative FSW. Conclusions: PrEP targeted to high-risk MSM or FSW in Bangalore could substantially reduce incidence amongst these groups, and have a small impact upon population HIV incidence, but may not be a very efficient use of resources in the short term. Higher efficiency may be achieved if PrEP is targeted to FSWs who are unable to use condoms consistently. 24

242 Posters Posters 20: PrEP: Resistance, Modeling and Acceptability P20.03 P20.04 LB Antiretroviral Pre-exposure Prophylaxis and Risk of Decreased Glomerular Filtration Rate among HIV Uninfected Men and Women Kenneth K. Mugwanya,2, Christina Wyatt 3, Connie Celum, Deborah Donnell,4, Nelly R. Mugo,5, James Kiarie 6, Allan Ronald 7, Jared M. Baeten, Partners PrEP Study Team University of Washington, Seattle, WA, United States, 2 Makerere University, Kampala, Uganda, 3 Mount Sinai School of Medicine, New York, NY, United States, 4 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 5 Kenya Medical Research Institute, Nairobi, Kenya, 6 University of Nairobi, Nairobi, Kenya, 7 University of Manitoba, Winnipeg, MB, Canada Background: Oral tenofovir disoproxil fumarate (TDF)-based preexposure prophylaxis (PrEP) protects against HIV acquisition. TDF use has been associated with declines in estimated glomerular filtration rate (egfr) when used as part of antiretroviral treatment by HIV infected persons but limited data are available for risk when used as PrEP by HIV uninfected adults, especially among those with high PrEP adherence. Methods: We conducted a safety analysis of the change in egfr in the Partners PrEP Study, a randomized, placebo-controlled trial of daily oral TDF and emtricitabine (FTC)-TDF PrEP among heterosexual HIV uninfected members of serodiscordant couples. PrEP adherence was high in the study population. egfr was calculated using the Chronic Kidney Disease Epidemiology Collaboration Equation. Results: A total of 463 subjects were randomized to once daily TDF (n=545), FTC-TDF (n=542), or placebo (n=544) and followed for a median of 8 months (IQR 2-27). 62% of subjects were male and the median age at enrollment was 35 years (range 8-64). Overall, mean egfr change from baseline that was attributable to PrEP compared to placebo was -.23 ml/min/.73m 2 (95% CI -2.06, -0.40; p=0.004) for TDF and -.58 ml/min/.73m 2 (95% CI -2.42, -0.73; p< 0.00) for FTC- TDF. The difference in egfr between PrEP and placebo appeared by 4 weeks after randomization, was stable through 2 months, and then gradually waned. There was no statistically significant increase in the incidence of a 25% decline in egfr attributable to PrEP compared to placebo: incidence rate per 00 person-years.00 for TDF,.4 for FTC-TDF, and 0.76 for placebo (adjusted Hazard Ratios.34, 95% CI 0.72, 2.50; p=0.36 for TDF and.45, 95% CI 0.80, 2.64; p=0.23 for FTC-TDF). Conclusions: In this large randomized, placebo-controlled trial, TDFbased PrEP was associated with a small although statistically significant decline in egfr. However, there was no significant increase in the risk of clinically relevant ( 25%) egfr decline related to PrEP. Long-term Risk of HIV Drug Resistance after PrEP Breakthrough Infection: Implications from a Mathematical Model of the HIV Latent Reservoir Anna Bershteyn, Philip A. Eckhoff Institute for Disease Modeling, Bellevue, WA, United States Background: Clinical trials of HIV pre-exposure prophylaxis (PrEP) have found little risk of HIV drug resistance after breakthrough infection, provided that the infection is detected and PrEP is discontinued in a timely manner. However, broad roll-out of PrEP in resource-limited settings cannot guarantee ideal monitoring for all patients. Also unknown is the transmissibility of PrEP-related drug resistance, which may differ from that of ART-related drug resistance due to transmitted founder virus dynamics. Methods: We developed a mathematical model that links PrEP adherence to the dynamic populations of drug-sensitive and drugresistant virus, distinguishing between replicating virus and the longlived latent reservoir. The model was parameterized for tenofovirbased oral PrEP with viral fitness and resistance pattern data for K65R. Novel model structures were explored to study transmitter founder virus dynamics. Results: The size and persistence of the modeled drug-resistant viral pool grew fastest during acute viremia, but continued to grow over the duration of undetected infection. After cessation of oral PrEP, the latent HIV reservoir harbored approximately double the proportion of K65R compared to the actively replicating pool. Host immunity, which influences viral load setpoint, modulated the proportion of K65R over an order of magnitude, as did patterns of adherence during PrEP use. Resistance acquired during early HIV infection could potentially elevate the transmissibility of resistance, which was seen when models assumed early establishment of a stable founder virus. Conclusions: The need for early detection of breakthrough infections during PrEP is expected to be highly heterogeneous, with the greatest need among patients with pre-existing immune impairment. Importance of adherence-driven pharmacokinetics implies that other PrEP methods, such as microbicides, may yield very different resistance patterns. More research is needed to understand transmissibility of drug resistance acquired during PrEP. 242 HIV Research for Prevention 204 HIV R4P

243 Posters 2: Product Acceptability and Adherence Wednesday, 29 October P2.0 Participation of Individuals from Fishing Communities in a Simulated Vaccine Efficacy Trial in Preparation for Future HIV Prevention Work P2.02 Adherence Improvement Innovations to Enhance Protocol Compliance and Vaginal Ring Adherence in a Phase III HIV Prevention Study in KwaZulu-Natal Gershim Asiki, Andrew Abaasa, Ubaldo Bahemuka, Jerry Mulondo, Freddie Kibengo, Eugene Ruzagira, Anatoli Kamali, Pat Fast 2 Medical Research Council/Uganda Virus Research Institute, Entebbe, Uganda, 2 International AIDS Vaccine Initiative, New York, NY, United States Background: As part of preparation for future HIV prevention efficacy trials evidence is needed to establish if fisherfolk can be enrolled and followed in vaccine research trial centre that is located outside their work place. We report participation of fisherfolk in a simulated vaccine trial using Hepatitis B vaccine as a substitute for HIV vaccine to evaluate participation in future trials. Methods: Volunteers at risk for HIV aged 8+ years from fishing communities 30-40km away from the MRC/UVRI clinical research center in South West Uganda were recruited. Those who had completed at least 3 months of follow-up in an open cohort and with no contraindications for Hepatitis B vaccine were vaccinated following the standard schedule (0,, 6 months, no placebo/control arm). Three days after each vaccination and at 3, 9 and 2 months after the first vaccination, volunteers were requested to return for procedures designed to mimic those of a clinical trial with a candidate vaccine. Results: Of 282 (27% women) enrolled and vaccinated from July 203-February 204. Per protocol vaccination compliance at months and 6 was 237/280 (85%) and 94/273 (7%) respectively; compliance including vaccinations received approximately 2 days outside the scheduled dates was 265/280 (95%) and 248/273 (9%) respectively. Over 85% of the participants returned for each of the day-3 postvaccination visits. The only factor associated with non-compliance to visits was living less than one year in the fishing site (ahr=3.6, 95%CI [.4-9.2]). Gender, age, education, occupation and marital status were not associated with compliance. We observed 0 new HIV infections in 3.6 PYO, giving an incidence of %CI (.7-6.0) for this cohort. Conclusions: We have observed high retention in a fisherfolk cohort enrolledin a simulated vaccine trial. For better retention, only individuals that have spent more than one year in the fishing community may be suitable for future trials. Chantel Friend, Philip Kotze, Annalene Nel 2 Qhakaza Mbokodo Research Clinic, Ladysmith, South Africa, 2 International Partnership for Microbicides, Cape Town, South Africa Background: Qhakaza Mbokodo research clinic (RC) is situated in Ladysmith, a small city in rural KwaZulu-Natal and is participating in a Phase III vaginal ring HIV prevention study. Observed protocol noncompliance and product non-adherence were highlighted as possible risk factors. With assistance from the WHO guidelines, RC specific adherence strategies were developed to improve and enhance compliance. Methods: A multi-dimensional strategy involving adherence improvement projects across all disciplines was developed to potentially identify a profile of presumed non-adherers, by means of a RC-developed protocol-specific scoring system. To gather more insight, participants were invited to provide feedback in small groups targeting adherence discussion topics. Individual educational counselling sessions and group motivational meetings were held. Socio-economic factors, mostly due to the local economy and declining work opportunities, were raised and addressed during participant home visits. Additionally, to address difficulties in accessing the RC, a transport system was arranged. The participant visit tracking system was re-evaluated and technology used to communicate with participants and track ongoing monthly retention strategies. Results: By using a multi-dimensional approach to improving adherence, general improvement in protocol compliance over time was observed, including less participants contributing to missed or late visits. A decline in trial discontinuations was experienced but the most common reason remains the local economic climate leading to participants seeking job opportunities in larger metropolitan areas. Conclusions: Adherence is a risk factor for all HIV prevention interventions which require a participant s active participation in product use. It is a complex issue which cannot be addressed by using a single modality and should be addressed continuously by a multi-dimensional approach

244 Posters Posters 2: Product Acceptability and Adherence P2.03 P2.04 Knowledge and Perceptions of Vaginal Microbicides among Healthcare Postgraduate Students at the University of Limpopo Mookho Malahleha, Kebogile Mokwena 2, Khatija Ahmed Setshaba Research Centre, Pretoria, South Africa, 2 University of Limpopo, Medunsa, Department of Public Health, Pretoria, South Africa Background: Women account for about 60% of the HIV infected population, which necessitates prevention mechanisms that specifically target women. There are currently robust research initiatives around vaginal microbicides as a potential HIV prevention tool for women. The CAPRISA 004 study is currently the only study that has shown partial effectiveness of vaginal microbicides to prevent HIV and STIs, and there are confirmatory studies that could change the microbicide research field. Emerging technologies like microbicides require positive beliefs and knowledge from healthcare workers to ensure successful implementation. A greater understanding of knowledge and perceptions among healthcare professionals will influence the pathway to microbicide development and possible future implementation. This study was conducted to assess the knowledge and perceptions of vaginal microbicides among postgraduate healthcare professionals at the University of Limpopo, Medunsa. Methods: A quantitative descriptive cross sectional study was conducted among 78 postgraduate students in the Masters of Public Health programme at the University of Limpopo, Medunsa Campus. A self-administered questionnaire was used as data collection tool. Results: About 90% of the respondents indicated an awareness of vaginal microbicides. A level of knowledge, however, was assessed by the number of correct responses on questions posed about vaginal microbicides. Whilst the overall knowledge was around 50%, 78.9% of respondents had positive perceptions about vaginal microbicides as a potential HIV prevention tool for women. There were no sociodemographic factors that were identified to influence the positive perceptions of vaginal microbicides. Conclusions: The study observed that postgraduate healthcare students at the University of Limpopo were fairly knowledgeable about vaginal microbicides and had positive perceptions about microbicides as a potential HIV/ STI prevention tool for women. Assessing the Impact of Message Framing on Microbicide Acceptability Elizabeth E. Tolley, Allison P. Pack, Sam Field 2, Elizabeth Ryan 3, Bockh Emily 4, Caroline Mackenzie 5, Alice Olawo 5, George Githuka 6 FHI 360, Social & Behavioral Health Sciences, Durham, NC, United States, 2 FHI 360, Quantitative Sciences, Durham, NC, United States, 3 FHI 360, Social Marketing and Communication, Washington, DC, United States, 4 FHI 360, Global Health, Population & Nutrition, Washington, DC, United States, 5 FHI 360, Country Office, Nairobi, Kenya, 6 National AIDS & STI Control Programme, Nairobi, Kenya Background: In Kenya, the largest proportion of new HIV infections is in stable couples, suggesting the need to carefully position new prevention products so women can use them. We developed and assessed the efficacy of a minimum package of materials aimed at increasing microbicide interest among prioritized audiences of women (aged 6-45) and men (8-45). Posters, TV and radio spots were framed in two ways. HIV-framed materials promoted microbicides as HIV prevention; non-hiv framed materials highlighted other benefits (i.e., increased intimacy) along with HIV prevention. Our study evaluated whether efficacy differed by framing overall and among target groups. Methods: We enrolled 606 women and 98 men in two Kenyan cities, randomizing them to receive information-only, or view either HIV-framed or non-hiv framed materials. We ran multivariable linear regression models to assess the effect of randomization group, age and relationship status on Microbicide Interest in Use (MIU, 7-item scale, alpha=.96) and Stigmatizing Attitudes towards Microbicides (SAM, 8-item scale, alpha=.83). Results: Women s mean age was 26.6 and men s was About 66% of participants were recruited from low-risk public spaces and 34% from bars and nightclubs. Most participants were married or had a regular partner; fewer had no regular partner or were not sexually active. Few participants had heard of microbicides before the survey. Overall, MIU was high and SAM low, with some gender differences. There was no overall effect of framing on these two outcomes, but some differences by relationship status; married men and women viewing non-hiv framed materials scored significantly higher on MIU and marginally lower on SAM than those viewing HIV framed materials. Conclusions: The way microbicides are framed does impact interest in use; different framing may be needed for different audiences. Microbicide introduction strategies should promote the product s non-hiv benefits in addition to HIV protection to achieve higher acceptability. 244 HIV Research for Prevention 204 HIV R4P

245 Posters 2: Product Acceptability and Adherence P2.05 Microbicide Counseling Tools for Health Care Providers: Promoting Product Acceptability and Adherence Wednesday, 29 October Allison P. Pack, Caroline Mackenzie 2, Alice Olawo 2, Elizabeth Ryan 3, Emily Bockh 3, George Githuka 4, Elizabeth E. Tolley FHI 360, Durham, NC, United States, 2 FHI 360, Nairobi, Kenya, 3 FHI 360, Washington, DC, United States, 4 Ministry of Health, Nairobi, Kenya Background: In 200 CAPRISA 004 provided proof of concept that peri-coital, vaginal use of tenofovir % microbicide gel can reduce HIV acquisition among women. Communication campaigns, including information for health care providers, will play a key role in promoting product acceptability and adherence. Communicating about Microbicides with Women in Mind developed and assessed a suite of materials for Kenyan audiences. The project is part of a larger USAID-funded effort to support microbicide introduction if/when tenofovir gel is proven effective. Methods: After receiving input from various stakeholder engagement activities, we worked with a local creative firm to design materials for health care providers, including a wall chart, counseling flipchart and brochure. We conducted 24 in-depth interviews (IDIs) from a range of public and NGO facilities in Nairobi and Nakuru to assess providers acceptability of and ability to appropriately use materials to counsel women on microbicides. IDIs explored providers thoughts on the content and usefulness of materials; scenarios were used to facilitate a mock counseling session. Results: Overall, providers had a positive view of microbicides, and felt materials were useful and could be incorporated into facility procedures. Most IDIs revealed preexisting rote counseling tendencies in which providers reported counseling couples to be faithful, female sex workers to use condoms, and single women to abstain. However, more effective counseling was demonstrated during the IDIs through the use of scenarios. Overall, providers felt that the microbicide counseling materials would help increase the range of prevention options they could provide to women. Conclusions: Health care providers will play a key role in promoting microbicide acceptability and adherence. Microbicide communication materials for providers were well-received and effective at helping counsel women in different sexual contexts. Provider training will be necessary to ensure appropriate material use

246 Posters Posters 22: Resistance and Treatment Failure P22.0 Prevalence and Correlates of Nevirapine (NVP) Resistance in NVP Unexposed HIV- Infected Infants Initiating Early Antiretroviral Therapy P22.02 Estimating HIV- Drug Resistance in Patients Experience Treatment Failure in Tegucigalpa- Honduras, during April 203 to March 204 Bhavna H. Chohan,2, Kenneth Tapia 2, Sarah Benki-Nugent 2, Musa Nga yo 3, Brian Khasimwa 4, Dara Lehman 5, Dalton Wamalwa 4, Julie Overbaugh 5, Grace John-Stewart 2 Kenya Medical Research Institute, Center for Virus Research, Nairobi, Kenya, 2 University of Washington, Seattle, WA, United States, 3 Kenya Medical Research Institute, Nairobi, Kenya, 4 University of Nairobi, Nairobi, Kenya, 5 Fred Hutchinson Cancer Research Center, Seattle, WA, United States Background: Nevirapine-based antiretroviral therapy (NVP-ART) is commonly used in resource-limited settings for treatment of NVP unexposed HIV-infected infants. However, the frequency of resistance and impact of the resistance on viral suppression in this group of infants initiating early ART, have not been well-characterized. Methods: To address this, we determined serial prevalence of NVP resistance during 2 months of ART among NVP-unexposed HIVinfected infants, enrolled in a clinical trial and initiated on NVP-ART, and determined effect of NVP resistance on viral levels. Results: Of 99 infants screened, 42 had no reported NVP exposure, and 22 infants with no baseline NVP resistance were initiated on NVP-ART. During follow-up, 7 infants (32%) developed resistance, infant at 3 months, and 6 at 6 months after ART initiation. HIV- RNA levels were similar at baseline among infants who developed resistance and those who did not (P=0.3). There was a trend for reduced viral suppression at 3 months of ART in infants with resistance than in those without (P=0.4) and after 6 and 2 months of ART, viral levels were significantly higher in infants with resistance than those without (P=0.007, P=0.04, respectively). Conclusions: Among infants without previous exposure to NVP development of NVP resistance was frequent, and presence of resistant mutations in these infants was significantly associated with less viral suppression during the first year of ART. High prevalence of NVPresistance during ART despite lack of NVP exposure may be due to higher baseline viral levels and longer viremia in infants following ART in contrast to adults. Development of NVP resistance may, in part, explain superiority of Protease inhibitor-based ART in infants. Wendy Murillo, Ivette Lorenzana de Rivera, Candy Carbajal, Erika Perez, Norma Solorzano 2, Elsa Palou 3, Elvia Ardón 4, Claudia García-Morales 5, Santiago Avila-Rios 5, Gustavo Reyes- Terán 5 Universidad Nacional Autónoma de Honduras, Escuela de Microbiología, Tegucigalpa, Honduras, 2 Instituto Nacional Cardiopulmonar, Infectología, Tegucigalpa, Honduras, 3 Hospital Escuela Universitario, Tegucigalpa, Honduras, 4 Secretaría de Salud, Departamento de ITS, VIH y SIDA, Tegucigalpa, Honduras, 5 Centro de Investigación en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias, Mexico City, Mexico Background: Current suppressive combination antiretroviral therapy (cart) has improved the health and life spam in HIV- infected patients. However, the long-term efficacy of cart has been compromised in many individuals, often due to the development of drug-resistance mutations (DRM). Surveillance of DRM in antiretroviral-experienced patients may provide useful information regarding rescue intervention. Methods: Plasma sample from HIV- infected patients on cart with signs of virological, and/or immunological and/or clinical failure were collected during April-203 to March-204, at the two principal HIV-care clinics in Tegucigalpa, Honduras. DRM analysis of HIV- pol sequences were obtain with the Stanford University Database. Results: A total of 65 specimens were analyzed. The proportion of patients with DRM was 75% (95% IC: 68-8%), 74% in adults and 92% in children. DRM were 85% for NRTI, 92% for NNRTI and % for PI. Overall, 2.% patients showed DRM to at least one drug class (6.5% for NRTI and 4.6% for NNRTI), 69.9% to at least two (67.5% for NRTI+NNRTI, 0.8% for NRTI+PI, and.6% for NNRTI+PI), and 9% to all three drug families (NRTI+NNRTI+PI). DRM was observed in 64% patients who were on first cart regimen, 26% on second, 9% on third, and one patient (0.8%) was on his seventh cart regimen. Conclusions: This is the second study of this kind, since cart has been implemented in the country in The rate of DRM has slightly declined (75%) in antiretroviral-experience patients with treatment failure compare with the rate reported in 2009 (8%). The majority of the patients showed single- and dual-class resistance. There has been a significant decreasing of multi-drug resistance from 33% reported in 2009 to 9% in 204 (p < 0.000). Resistance testing implementation is an urgent need in the country because the management of heavily treatment-experience HIV-infected patients represents a considerable challenge for HIV-care clinicians. 246 HIV Research for Prevention 204 HIV R4P

247 Posters 22: Resistance and Treatment Failure Wednesday, 29 October P22.03 Effectiveness of Drug Interventions Reducing Nevirapine Resistance after Single-dose Exposure for Perinatal HIV Prevention Eva P. Muro, Quirine Fillekes 2, David M. Burger 2 Kilimanjaro Clinical Research Institute, Moshi, Tanzania, United Republic of, 2 Radboud University Nijmegen Medical Centre, Department of Pharmacy, Nijmegen, Netherlands P22.04 High Prevalence of Cross-resistance to Rilpivirine in Subtype C HIV- Isolates from First-line ART Failures in South Africa Kerri J. Penrose, Carole L. Wallis 2, Maritsa Scoulos-Hanson, Raquel Viana 2, John W. Mellors, Urvi M. Parikh University of Pittsburgh School of Medicine, Infectious Diseases, Pittsburgh, PA, United States, 2 Lancet Laboratories, Specialty Molecular Division, Johannesburg, South Africa Background: The major disadvantage of single-dose nevirapine is the development of nevirapine resistance in mothers and infants. Co-interventions, such as short-course antiretroviral regimens in combination with single-dose nevirapine and extension of interventions, have been evaluated in this study. Methods: Systematic search of electronic databases (MEDLINE, EMBASE and Cochrane) was performed. Studies included HIV-infected, pregnant women, who were administered single-dose nevirapine for pmtct and who were receiving an intervention to reduce nevirapine resistance. Primary outcome was the proportion of nevirapine resistance detected in plasma samples collected 3 months postpartum. The reducing effect of drug interventions on nevirapine resistance was assessed in metaanalyses using random effects models and the GRADE approach for quality of evidence. Results: The estimated pooled proportion of nevirapine resistance using single-dose nevirapine at labor was 3% (95%CI ); this was reduced to 2% (95%CI ) with addition of antepartum zidovudine. A combination of antepartum zidovudine, single-dose nevirapine and a short (< 8 days) postpartum regimen resulted in a major reduction in nevirapine resistance to 0.0% (95%CI ). The summary effect of days of postpartum drug regimens combined with antepartum zidovudine and single-dose nevirapine was associated with a slightly lower incidence of nevirapine resistance, namely 0.003% (95%CI ). Conclusions: Antepartum zidovudine plus antiretroviral drugs postpartum have shown to nearly eliminate nevirapine resistance. Although days post partum regimens might be slightly more effective compared to a short (< 8 days) postpartum regimen, longer term antiretroviral therapy is more complex and more challenging to implement in daily practice. The WHO guideline option A should be followed to achieve a feasible minimum risk of nevirapine resistance in regions where single-dose nevirapine is still being used. Background: TMC278LA (RPV) is a promising PrEP agent due to its subnanomolar potency and injectable, long-acting formulation, but the rise in transmitted NNRTI resistance has the potential to reduce RPV s preventive efficacy. This study investigated RPV cross-resistance among recombinant subtype C viruses derived from individuals failing first-line NNRTI-containing regimens in South Africa (SA). Methods: Plasma samples were collected prospectively from HIV- subtype C-infected individuals who were: i) failing first-line ART after 6 months of treatment; ii) had plasma HIV RNA >0000 copies/ml; iii) had NNRTI-resistance mutation in RT. Recombinant HIV- LAI containing bulk-cloned full-length RT sequences from 88 plasma samples were assayed for NVP, EFV and RPV susceptibility in TZM-bl cells. Fold-change resistance (FC) was calculated against a mean IC 50 from 3 subtype C HIV- samples from SA. To relate in vitro IC 50 values to human plasma protein adjusted IC 90 values and achievable RPV concentration in vivo, we set the cutoff for RPV resistance at >42.6 ng/ml based on the average C min for RPV following successive 200/600/600 mg monthly TMC278LA injections in clinical studies. Results: 37/88 (42%) samples showed resistance to RPV defined by FC>0 and an adjusted IC 90 >42.6 ng/ml. 36/37 (97%) samples were from failures of EFV-containing ART and from NVP-containing ART. The 3 most frequently detected NNRTI mutations among the 37 RPV-resistant viruses were K03N (68%), L00I (30%) and Y8C (22%), compared to V06M (56%), G90A (35%) and K03N (33%) in susceptible samples. Conclusions: Frequent cross-resistance to RPV was observed among HIV- subtype C viruses from individuals experiencing failure of firstline NNRTI-containing ART. The level of RPV cross-resistance for 42% of viruses exceeded the expected plasma C min for RPV following successive monthly IM doses of TMC278 (200/600/600 mg), suggesting that transmission with NNRTI-resistant HIV- could occur with TMC278LA administered with this dosing regimen

248 Posters Posters 22: Resistance and Treatment Failure P22.05 Correlates of Adherence and Antiretroviral Treatment Failure in HIV- Infected Kenyan Patients Washingtone Ochieng,2, Rose C. Kitawi,3, Timothy J. Nzomo,3, Ruth S. Mwatelah,3, Charity Hungu 2, Maureen J. Kimulwo,3, Kevin O. Onyango,4, Raphael M. Lwembe 2, Mkaya Mwamburi 5, Berhnards R. Ogutu,6, Florence A. Oloo,7, Rashid Aman,8 Center for Research in Therapeutic Sciences, Nairobi, Kenya, 2 Kenya Medical Research Institute, Centre for Virus Research, Nairobi, Kenya, 3 Institute of Tropical Medicine and Infectious Diseases, Nairobi, Kenya, 4 Novartis Vaccines and Diagnostics Inc., Cambridge, MA, United States, 5 Tufts University School of Medicine, Center for Global Public Health, Boston, MA, United States, 6 Kenya Medical Research Institute, Centre for Clinical Research, Nairobi, Kenya, 7 Kenya Technical University, Nairobi, Kenya, 8 African Centre for Clinical Trials, Nairobi, Kenya Background: Universal access to antiretroviral treatment (ART) is still elusive in most developing nations. We report on the effectiveness of the current ART scale-up campaign in Kenya and discuss factors influencing treatment outcomes. Methods: A multi-center longitudinal and cross-sectional survey of viral load (VL), CD4 T-cells, drug resistance and adherence. VL, CD4 counts and drug resistant mutations were determined using m2000 Abbott HIV- assay, FACS and Pol-RT sequencing respectively. Adherence was scored as good, fair or poor based on number of missed doses. Results: Overall, 35.9% of the 546 patients failed treatment using longitudinal multiple viral load (VL), and 22% to 29% failed using cross-sectional single-vl definitions. More patients (4%) starting first-line D4T+3TC+NVP/EFV failed treatment than those initiating TDF+3TC+NVP/EFV (29%) (P=0.043). Female patients had higher CD4 counts, lower VL, better adherence and significantly less ART failure than males. Using Chi-Square test, the cross-sectional criteria defined failure with 99% to 93% accuracy of the longitudinal VL approach (p< 0.00). Patients switched regimen without necessarily failing first-line, and 26% of those switching still failed second-line. Up to 33% of the patients had at least two major drug resistance mutations of NRTI or NNRTI type. Community Peer Support Network (CPSN) activity was significantly associated with improved adherence, reduced VL and reduced treatment failure rates. Patients with multiple active sex partners also had higher VL, but these were not significantly different between independent groups. Conclusions: D4T-containing primary regimen and weak adherence independently correlate with increased ART failure. A single VL test after 2 months of uninterrupted ART offers an effective alternative to ART failure definition under limited resources. We recommend point-of-care VL testing at least once annually, drug resistance monitoring and peer focused adherence-support to inform treatment and mitigate failure. 248 HIV Research for Prevention 204 HIV R4P

249 Posters 23: Retention and Adherence in Trials Wednesday, 29 October P23.0 Frequency of Multiple Sexual Partnerships among Women Participating in an HIV Prevention Trial in Durban, South Africa P23.02 The Relationship between Stigma, Disclosure, and Adherence among Participants in the African Cohort Study Zakir Gaffoor, Renee Street, Nathlee Abbai, Handan Wand 2, Gita Ramjee Medical Research Council of South Africa, HIV Prevention Research Unit, Durban, South Africa, 2 The Kirby Institute, University of New South Wales, Sydney, Australia Background: In 202, an estimated 35.3 million people were living with HIV/AIDS. Sub-Saharan Africa accounts for 70% of all new infections with women being disproportionately affected. Risky sexual behaviour practices are contributory to the high HIV prevalence in this population. The aim of this study was to describe a cohort of women enrolled in an HIV prevention trial who have multiple sexual partnerships. Methods: Self-reported data was obtained from intervieweradministered sexual behaviour and demographic questionnaires of 485 women that participated in the Carraguard vaginal microbicide study. In this report, a cross-sectional analysis of women who reported having one steady sexual partner (366/485) compared with women who reported having additional sexual partners over and above a steady sexual partner (9/485), was conducted, with the outcome variable being self-reported multiple sexual partnerships. Results: A total of 485 women were included in the analysis. Of these 9 (8 %) of them reported at least one other sexual partner besides their current regular partner. Overall less than 50% of the women reported using condom at last sexual act. Younger women (< 25 years old) were more likely to report multiple partners, compared to women aged and over 30 (p=0.00). Other factors that were associated with women engaging in multiple partnerships included other high risk behaviours such as reporting ever experiencing forced sex; ever having sex for cash; ever experienced physical abuse (p< 0.00 all) and at least one form of contraception use at screening (p=0.044). Conclusions: The prevalence of multiple sexual partnerships reported by women in this study is similar to other studies conducted in Africa. Our results therefore provide further impetus for targeted HIV prevention messages in addition to female controlled methods for HIV prevention. Lindsay Hughes,2, Tiffany Hamm,2, Samoel Khamadi,3, Lucas Maganga 4, Hannah Kibuuka,5, Francis Kiweewa,5, Ogbonnaya Njoku,6, Babajide Keshinro,6, Jonah Maswai,7, Appolonia Aoko,7, Milton Omondi,8, Nekoye Otsyula,8, Christina Polyak,2, Merlin Robb,2, Nelson Michael, Julie Ake U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Bethesda, MD, United States, 2 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 3 Walter Reed Program Tanzania, Mbeya, Tanzania, United Republic of, 4 Mbeya Medical Research Centre, National Institute for Medical Research, Mbeya, Tanzania, United Republic of, 5 Makerere University Walter Reed Project, Kampala, Uganda, 6 Walter Reed Program Nigeria, Abuja, Nigeria, 7 Walter Reed Project Kenya, Kericho, Kenya, 8 Walter Reed Project Kenya, Kisumu, Kenya Background: Stigma is recognized as a barrier to HIV prevention, contributing to decreased uptake of prevention and treatment services. Despite considerable literature on its impact, there is an evidence gap about discrete effects on disclosure and treatment adherence behaviors. Methods: Participants were recruited from client lists into the African Cohort Study (AFRICOS) at MHRP PEPFAR-supported clinics in Kenya, Nigeria, Tanzania, and Uganda. Questionnaires administered at enrollment related to stigma, disclosure, and adherence were analyzed for HIV+ participants. Results: The cohort (N=596) is 88% HIV+, 60% female with a median age of 39. Participants disclosure patterns appear related to perception of blame for HIV acquisition (e.g., 48% of those reporting HIV infection via sex with a regular partner report that their spouse/partner was aware of their status, compared to those acquiring HIV via forced sex or blood transfusion [75% and 00%, respectively]). Reported days with missed ARVs in the month prior to enrollment is significantly associated with stigmatizing experiences, specifically social isolation (p< 0.0), physical violence (p< 0.05), and broken family relationships (p< 0.05). There is a significant correlation between participants attributing missed doses to stigma-avoidance and days with missed ARVs during the preceding month (p< 0.0); no experience with HIV-related stigma was significantly negatively correlated with days with missed ARVs (p< 0.0). Participants who reported that none of their family or community members knew their HIV status were significantly more likely to report missing a clinic visit due to HIV-associated stigma (p< 0.05). Conclusions: Stigma, including self-stigma, is related to missed ARVs and disclosure behaviors. Participants who have not disclosed their status are likely to miss clinic visits and/or treatment doses to avoid revealing their status, suggesting stigma continues to be a barrier to effective treatment and successful execution of the HIV care continuum

250 Posters Posters 23: Retention and Adherence in Trials P23.03 Performance of the Wisebag for Monitoring Daily Rectal Gel Application in Two Urban Cities in the United States P23.04 Implementation of Tools to Optimize Protocol Visit Compliance at the Madibeng Centre for Research (MCR) Cindy E. Jacobson, Raymond Cefola 2, Amy Herrick 3, Rita Labbett, Elena Khanukkova 4, Ross Cranston 5, Peter Anton 4, Ian McGowan 5 Magee Womens Research Institute, Pittsburgh, PA, United States, 2 Magee Womens Hospital, Pittsburgh, PA, United States, 3 Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA, United States, 4 David Geffen School of Medicine at UCLA, Los Angeles, CA, United States, 5 University of Pittsburgh, Department of Medicine, Pittsburgh, PA, United States Background: Gel adherence is an issue that plagues microbicide trials. Electronic monitoring devices are thought to provide more detailed and accurate data on study product use. Methods: In an effort to obtain an assessment of adherence in a randomized, double-blind trial comparing three formulations of % tenofovir gel administered rectally, 2 participants received their applicators in a Wisebag. The Wisebag is a lunch-bag style container which contains a device with a communication chip which transmits a message to a server every time the Wisebag is opened. These stored data were compared to participant report of gel administration as well as applicator (used and unused) counts. Participants were to insert the rectal gel daily for five consecutive days for each of three different formulations. Results: Participants at both sites indicated on self-report that they administered the five consecutive doses of gel for each of the three formulations. This was also reported by count of used and unused applicators. These reports could not be supported or refuted by the data provided from the Wisebag recordings. The Wisebag data indicated that for all participants the bag was opened more than once on at least one day up to more than once daily for all 5 days. In addition to difficulty interpreting the frequent recorded openings the bags were fraught with technical challenges throughout the trial. Conclusions: Although the Wisebag may become an important measure of adherence in the future, logistical problems associated with this technology need to be addressed. Although it is understood that there is no way to control the frequency participates open the bag despite counseling, the Wisebag performance fell short of its anticipated usefulness. Cheryl Emily Louw, Marthie de Villiers, Hanlie Veldsman, Ntswaki Rose Masilo, Nicole Dippenaar, Sieglinde Kruger, Annalene M. Nel 2, Michelle Isaacs 2, Colleen Herman 2 Madibeng Centre for Research, Brits, South Africa, 2 International Partnership for Microbicides, Paarl, South Africa Background: MCR began its participation in a Phase III microbicide vaginal ring trial early in 202. Due to the high recruitment target and monthly scheduled visits, the research centre (RC) developed an Excel tracking sheet for participants and their visits. However, despite these efforts missed and late visits were still experienced over the 202 Festive Season. To follow-up and comply with protocol visits, the RC needed to improve its tracker to identify potential late visits earlier. Methods: The RC team: Refined the visit tracker to identify missed visit dates by highlighting visit window periods Management intensified their supervision of visit tracking with real time communication to the recruitment and retention team, with assistance from the Community Liaison Officer (CLO) Identified a dedicated Retention Officer to contact participants who missed their target date Emphasized the importance of active retention efforts to the recruitment and retention teams Results: Missed and late visits reduced and remained sustained at a much lower level, even over the 203 Festive Period. The recruitment and retention team maintain high vigilance for participants who tend to be late attendees, ensuring they understand the importance of complying with the protocol and attending visits within their visit windows. Conclusions: Appropriate trackers are very useful tools in enhancing follow-up for protocol compliance and ensuring visits occur on time. Dedicated team members should be utilized to encourage participants to return for their visits on time. Protocol compliance efforts require leadership of the Principal Investigator and co-investigators to sustain the effort, as this remains a team effort. 250 HIV Research for Prevention 204 HIV R4P

251 Posters 23: Retention and Adherence in Trials Wednesday, 29 October P23.05 Socio-demographic Characteristics of Female Sex Worker s in the Inner-city Johannesburg Hillbrow Goitse Manthata, Jabhile Maria Sibanyoni, Nyaradzo Mutanha, Petros Jacob Motsamai, Willem D.Francois Venter Wits Reproductive Health & HIV Institute, Sex Workers and Truckers Programme, Johannesburg, South Africa P23.06 Participant Deception: A Case Study in CAPRISA 008 Kathryn T. Mngadi, Nomzamo Mvandaba, Nonhlanhla Langa, Ntombifuthi Mkhize, Charlene Harichund, Quarraisha Abdool Karim,2, Leila E. Mansoor Centre for the AIDS Programme of Research in South Africa, HIV Prevention, Durban, South Africa, 2 Mailman School of Public Health, Columbia University, Department of Epidemiology, New York, NY, United States Background: Sex workers have been identified as one of the key population groups in terms of public health and HIV prevention programmes, and identified as a potential priority group for antiretrovirals treatment and pre-exposure prophylaxis, but little is known about this stigmatized group. Since 996, the Wits Reproductive Health and HIV Institute s Sex Workers Programme, in inner-city Johannesburg, has been providing health services to sex workers. Methods: Descriptive longitudinal data analysis of clinical data was conducted from November 20 - March 203 Results: Among the 2, 03 female sex workers, the majority (48%) were aged 28-38, most (49%) had obtained at least a grade 8-0 education and 82% were single (not married and did not have a main partner). The analysis also found that the large majority (86%) of the female sex workers were from Zimbabwe; 49% of those from South Africa were from Kwa-Zulu-Natal. Condom use was found to be most frequent with clients (99.45%), less frequent with casual partners (62%) and least frequent with main partners at 39.03%. Although the frequency of condom use was high amongst female sex workers and their clients, around 55% had an active STD. More specifically, the HIV prevalence was found to be around 60%, of those who tested and 43% (N=95) were on ART. Conclusions: Sex workers had very high condom use with clients, but lower condom use with casual and main partners; less than half of female sex workers were on ART, but this may reflect the current starting CD4 threshold of 350 cells/ul. HIV positivity levels are high, and focused ART programmes, including PrEP, seem appropriate for this group as both a public health and personal health measure. Background: Participant safety and data integrity, critical in trials of new investigational drugs, are achieved through honest participant report and fidelity in the conduct of procedures. Co-enrolment by professional participants is an example of participant deception, which has the potential to compromise participant safety and data integrity. We describe a case study of participant concealment of pregnancy to fulfil trial eligibility in order to access trial product, risking the safety of her unborn baby. Methods: The participant was enrolled in April 203 and attended four monthly follow-up visits each with negative urine pregnancy tests. Selfreport and returned used applicator counts at all visits indicated more than 90% product adherence. She received two depot medroxyprogesterone acetate injections. The pregnancy was detected clinically at the fourth visit during a clinically-indicated pelvic examination, and confirmed on ultrasound and blood pregnancy testing. A root cause analysis by the senior protocol team (investigators, clinicians, quality manager and bioethicist) examined pregnancy test kit integrity, user competence, clinical acumen of research staff and veracity of participant report. Results: The major finding, based on participant self-report, was that of participant deception, including denial of the pregnancy at screening, deliberate substitution of her urine sample with that of another person to ensure a negative pregnancy test result, and ignoring the product safety education provided by the team. She reported a desire to access HIV prevention during an unplanned pregnancy with a partner whose fidelity was in question. Unverified concerns remain around sharing/ selling of the product. Conclusions: Researchers need to balance systems put in place to counter the low prevalence of deception with the suspicion placed on the majority of honest participants. 25

252 Posters Posters 23: Retention and Adherence in Trials P23.07 P23.08 Assessment of Retention Rates using Mobile Phones versus Physical Contact Tracing among Fishing Communities along the Shores of Lake Victoria, Uganda Juliet Mpendo, Annet Nanvubya, Ali Ssetaala, Julius Ssempiira 2, Annet Nalutaaya 2, Matthias Wambuzi, Leslie Nielsen 3, Steven Asiimwe 4, Noah Kiwanuka UVRI-IAVI HIV Vaccine Program, Clinical & Epidemiological Sciences, Entebbe, Uganda, 2 UVRI-IAVI HIV Vaccine Program, Data Management, Entebbe, Uganda, 3 International AIDS Vaccine Initiative, New York, NY, United States, 4 Kabwohe Clinical Research Center (KCRC), Kabwohe, Uganda Background: More HIV prevention vaccine efficacy trials will have to be conducted in the future, and high participant retention rates are key characteristics of populations suitable for such trials. We conducted a study to assess study retention by comparing use of mobile phone reminders with physical contact tracing. Methods: A total of 662 participants aged 3-49 years from four fishing communities, screened for high risk behaviours were enrolled and randomized : to mobile phone arm where they were reminded by a phone call (33) or physical tracing arm (33). Participants were followed up mimicking a vaccine efficacy schedule at 0,, 2, 3,6,2 and 8 months. At each visit a semi-structured questionnaire was administered on socio demographics, risk behaviors for HIV and STIs, and treatment seeking behavior. Participants were reminded about an upcoming visit by a phone call or physical contact tracing, 7 days before and 3 days past the scheduled visit if by that time they had not turned up. Each visit had a window of +/- week. Retention was determined and compared across study arms. Data presented is up to month 2. Results: Overall the physical tracing arm had better retention rates compared to the phone but significant differences were only observed at month 2. Month, (79.5% vs 78.3%, respectively) month 2, (87.6% vs 87.3%), month 3, (86.7% vs 83.4%), month 6, (87.4 vs 85.5%) with the highest rate at month 2, (9.5% vs 82.5%; p- value = 0.00). Total compliance at all visits by month 2 was 59.4% overall and there was no significant difference between arms (57.7% vs 6.0%). Factors significantly associated with retention were resident on island (54.8% vs 64.6%) p value 0.0, tribe and attainment of secondary education. Conclusions: In fishing communities around Lake Victoria, Uganda, high retention rates can be achieved, with physical tracing appearing to yield higher study retention rates than mobile phone reminders. Motivating, Measuring and Monitoring Adherence in the FACTS 00 Tenofovir Gel Microbicide Study Sandra Mudhune, Sinead Delany-Moretlwe, Deborah Baron, Sinazo Pato, Jonathan Stadler, Nkosinathi Ngcobo, Mamakiri Khunwane 2, Sylvia Makgopa 3, Matshidiso Malefo 3, Hilda Ntjana 4, T Nkompela 4, Gugu Sibisi 5, T Ikaneng 6, K Seemise 6, Sanele Mdanda 7, K Manenzhe 8, M Morolo 8, Glenda Gray 2, Helen Rees Wits Reproductive Health and HIV Institute, University of the Witwatersrand, Johannesburg, South Africa, 2 PHRU, Johannesburg, South Africa, 3 Aurum Institute, Johannesburg, South Africa, 4 Desmond Tutu HIV Foundation, Cape Town, South Africa, 5 MatCH Research (Maternal, Adolescent, and Child Health Research), Pietermaritzburg, South Africa, 6 Mecru Medunsa University, Pretoria, South Africa, 7 Qhakaza Mbokodo Research Clinic, Ladysmith, South Africa, 8 Setshaba Research Centre, Pretoria, South Africa Background: Achieving optimal adherence is a challenge to evaluating oral and topical PrEP. Based on lessons from previous trials, we developed a package of interventions in FACTS 00 to address adherence barriers. FACTS 00 is a phase III licensure trial of tenofovir % gel assessing safety and effectiveness in preventing HIV and HSV-2 in women when used pericoitally. We report on feedback from staff implementing these adherence interventions. Methods: We developed tools to identify adherence issues and give monthly feedback to participants: Returned used applicators were visually inspected for evidence of vaginal insertion; a checklist assessed missed visits, relationship status, and gel use; an Excel tool calculated adherence levels by dividing half the number of used gel applicators by the number of reported sex acts. Additional interventions aimed to instil positive social norms included posters, motivational SMS, speeches by former trial participants, and a barometer of average adherence levels. Results: These tools helped refine counselling messages, target interventions at specific participants, and feedback individuals adherence progress over time. The positive norms campaign used multiple methods to promote regular gel use. The images and messages resonated with the everyday lives of South African women, recognizing the challenges of incorporating gel use, and suggested ways of overcoming these challenges. Intensive training, monitoring and support at site visits are essential to ensure that staff is able to implement these approaches to promoting adherence. Conclusions: Honing in on core motivators and barriers of adherence and leveraging positive peer opinion, we attempted to build positive trial norms to optimise adherence to gel. It has been essential to use tools implemented at an individual level to identify participants that need more support combined with campaigns aimed at creating a positive image of the gel and a supportive context, particularly when contextual factors influence gel use. 252 HIV Research for Prevention 204 HIV R4P

253 Posters 23: Retention and Adherence in Trials Wednesday, 29 October P23.09 Maximizing Participant Retention in a Phase2 B HIV Prevention Trial in Kampala, Uganda: The MTN-003 (VOICE Study) Rosemary Muwawu, Flavia M. Kiweewa,2, Fred B. Kapaata, Florence Kikonyogo, Regina Bukenya, Jane Musisi, Mary Kaggwa, Margaret Saava, Samuel Kabwigu, Clemensia Nakabiito Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 Makerere University School of Public Health, Makerere College of Health Sciences, Kampala, Uganda P23.0 Recruitment for Retention in Biomedical HIV Prevention Studies: Strategies, Challenges, Lessons Learned from MTN-020 (ASPIRE) Study, at Kampala Site Sophie Clare Nanziri, Patrick Ndawula 2, Teopista Nakyanzi 2, Brenda Gati 2, Flavia K. Matovu 2, Juliane Etima 2, Samuel Kabwigu 2, Doreen Kemigisha 2, Stella Nanyonga 2, Cleemensia Nakabiito 2 Makerere University - Johns Hopkins University Research Collaboration, Social Support, Kampala, Uganda, 2 Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda Background: The success of longitudinal trials depends greatly on effective strategies to retain participants to ensure internal validity. This paper describes the challenges, and strategies in retaining participants in the MTN-003 (VOICE) trial at Makerere University-John Hopkins University Collaboration, Uganda. Methods: A total of 637 women aged 8 to 45 were screened. Once enrolled, participants were seen every 28 days for HIV testing and product refill among other procedures. Retention and visit schedule adherence were critical due to short visit windows. Challenges to good retention included mobile population, poor communication, non-disclosure to family, and economic constraints. Strategies to maintain participation rates included adherence counseling, use of locator information, a tracking database, medical care, and close bonds between staff and participants. Participants were traced by health visitors if they did not come for their visit. Non-adherent participants were scheduled early to allow time for tracing. Results: Of the 637 screened, a total of 322 were enrolled. The overall retention rate was 95%. Only 79 (3%) of the 624 total visits expected were missed. Reasons for missed visits included being HIV negative and therefore thinking they did not need frequent visits, finding it difficult to attend visits due to sex work, and migration for better employment. There were a total of 8 early terminations; 3 withdrew consent, 9 were lost to follow up; 5 lost interest and died from a road traffic accident. Conclusions: With the implementation of comprehensive follow-up and retention strategies, high retention rates were achieved. These low technology, labor intensive methods are effective in a low resource setting. Background: Recruitment of participants is labor intensive and a critical aspect of prevention research. It is important to incorporate early retention techniques into recruitment strategies during the planning phase to ensure that retention targets are met. The Kampala team describes the strategies, process, and challenges in ensuring retainable participants are recruited into the study. Methods: Various strategies have been designed to address recruitment for retention. Participants are identified by community contact persons, after which recruitment staff use pre-screening checklists to identify those who would be retained. This is done in two phases; in the community and at the clinic before screening to check for consistency in participant responses. Critical indicators considered include: having stayed in an area for 6 months; willingness to provide adequate locator information, access to a reliable phone for easy tracing; stable jobs like the self-employed ones; positive health seeking behavior like initiating a modern contraceptive method, and interest in getting an HIV test before reporting for screening. At clinic Level, the main indicator is maintaining consistency of information using the pre-screening check lists that is administered in the community, and attempting to communicate with staff in case one fails to make it for their appointment. Results: Lessons learnt and challenges include; screening many women, making multiple contacts to get a few good ones. Some get to know what we want and coach others in the community. Those interested may lack public transport to get to the clinic or may not have a phone with good network to call for getting directions and rescheduling clinic visit appointment. To facilitate them, a site vehicle picks them from the community on the day of appointment. Conclusions: Overcoming recruitment and retention barriers involves planning and adequate recruitment strategies long before the study begins so as to recruit participants who are likely to be retained

254 Posters Posters 23: Retention and Adherence in Trials P23. P23.2 The Impact of Participant Mobility on Missed Visits among Women Taking Part in the VOICE (MTN-003) Study in Zimbabwe Pepukai Ndadziyira, Shingirayi I. Samupindi, Christine Vuta, Angeline Matanhire, Emilder Chihota, Patricia M. Dhlakama, Petina Musara, Margaret Mlingo, Nyaradzo M. Mgodi, Zvavahera Mike Chirenje University of Zimbabwe - University of California San Francisco Collaborative Research Programme, Harare, Zimbabwe Background: Effectiveness trials depend on good retention rates to minimize bias resulting from loss-to-follow-up. VOICE, a multi-site HIV prevention study was conducted in South Africa, Uganda and Zimbabwe. We identified the major driver of missed visits and demographic characteristics of women who missed visits in VOICE in Zimbabwe in an effort to give recommendations for accrual and retention strategies in future PrEP trials. Methods: Data of the 630 women enrolled at the 3 sites in Zimbabwe were analyzed. We used descriptive analyses to summarize demographic characteristics of the women who defaulted or missed their clinic visits. Results: A total of 24 women who had home visits after either defaulting or missing their scheduled visits were included in the analysis. These women contributed to 825 (87%) of the 949 missed visits reported in Zimbabwe. Average age of the 24 women was 27 years ( range 8-39) and average parity was 2. (range 0-5). The majority of women, 52 (94%) were married and 5 (93%) were currently staying with partner. A total of 93 (57%) worked as vendors and all, (00%) received financial support from partner and 94 (44%) owned their homes. All women had attained at least 7 years of primary school with 42 (88%) having completed 4 years of high school. Of the 825 missed visits, 662 (80%) were due to mobility as reported in 62 (76%) of the 24 women. Of the 62 women, 93(57%) missed due to rural visits, 34(2%) were employed in neighboring countries, 25(5%) were cross-border traders and 9(6%) had relocated to another city. The remaining 20% (63/825) of missed visits were due to partner influence reported in 8(4%) women, work/business commitments in 2(0%) and no reason stated in 2 (6%). Conclusions: The major driver of missed visits in VOICE in Zimbabwe was participant mobility. Future studies recruiting from catchment areas similar to VOICE should have more stringent measures to ascertain risk of possible mobility prior to enrolling women. Reaching Vulnerable MSM through Index Client Strategy Kennedy Otieno Olango Men Against AIDS Youth Group, Kisumu, Kenya Background: Research documents high HIV prevalence among young gay men, other men who have sex with other men (MSM) and transgender individuals (GMT) and reaching this highly vulnerable population in Kenya is hard. Our hypothesis is that empowerment of GMT testing positive to disclose their status to their partners and families and encourage them to go for testing as well can reduce their fear of isolation, increase their capacity for treatment access and retention, increase awareness among their colleagues, and inform development of appropriate services Methods: A group of 20 GMT+ who were enrolled on treatment and care were classified as index client and were used to reach other, their partners as friends. They were trained as on prevention with positive in regards to Positive, Health, Dignity and Prevention (PHDP) and formed an outreach team to conduct community PWP sessions among other fellow GMT /partners. They conducted health talk s forums on benefits of testing and treatment to their partners and peer and visited several support groups, raised visibility of HIV-positive GMT and advocated for treatment as prevention. Results: Reach MSM through Index Client strategy created spaces for GMT+ in to reduce isolation, increased capacity for health and represented their own HIV care and treatment needs. Programmatic evaluation indicated increased knowledge in PWP concepts, including status disclosure, partner testing, and STI and other infectious disease screening, and benefits of adherence, etc. Qualitative evaluation at the end of the campaign showed increased knowledge and practice related to status disclosure, partner testing and adherence Conclusions: Empowerment of GMT +and followed by treatment literacy campaigns has demonstrated opportunity to empower GMT +to raise awareness about HIV/AIDS, combat stigma ad isolation, and inform development of services and holistic approaches for the health of GMT+and help in reaching other GMT who are vulnerable or already positive in Kisumu County. 254 HIV Research for Prevention 204 HIV R4P

255 Posters 23: Retention and Adherence in Trials Wednesday, 29 October P23.3 P23.4 The Implementation of Group Education Sessions for Women Participating in HIV Prevention Research Studies in Durban, South Africa Samantha S. Siva, Jayajothi Moodley, Vaneshree Govender South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: The target population of women participating in HIV prevention studies in South Africa often lack information about health and socio-economic issues that impact their lives at an individual and community level. Women from similar backgrounds waiting to be seen at clinical research sites provided an opportunity for the implementation of group education sessions. This created a platform to discuss health and socio-economic factors and provide key study related messages to participants. The objective of this report is to describe how these sessions aimed to facilitate open discussions, encourage honest reporting of issues and dispel misconceptions. Methods: A participant centred approach is adopted whereby the sessions are directed by participants who suggest topics of interest such as sexually transmitted infections and treatment, contraception, drug abuse, gender based violence and topics related to current issues. Peer educators, who are previous or current study participants, are sometimes invited as facilitators. Contact details for community resources that could assist with challenges are provided. Participants also share their experiences with study participation and study product use. Results: The group education sessions allowed participants access to socio-culturally relevant information. Good rapport was developed between participants and staff and amongst participants themselves which contributed to interactive discussions. Women openly discussed challenges, such as disclosing study participation and product use to family members as well as myths regarding contraception and cervical cancer. Strategies to overcome these challenges were also shared. Conclusions: The implementation of group education sessions at clinical research sites has the potential to enhance commitment to study participation and provide a platform to address health and socioeconomic concerns. These women may act as a conduit for messages to the greater community and impact on public health. Acceptability of Financial Incentives for HIV Viral Suppression: A Qualitative Substudy of HPTN 065 Allison P. Pack, Jill Stanton, Elizabeth Greene, Jamilah Taylor, Victoria Shelus, Elizabeth E. Tolley, Natella Rakhmanina 2, Wafaa El-Sadr 3,4, Theresa Gamble, for the HPTN 065 Study Team FHI 360, Durham, NC, United States, 2 Children s National Medical Center, Washington, DC, United States, 3 Columbia University Mailman School of Public Health, ICAP, New York, NY, United States, 4 Harlem Hospital, New York, NY, United States Background: HPTN 065 (TLC-Plus) studied the effect of providing quarterly $70 financial incentives (FI) to HIV-infected patients on antiretroviral therapy (ART) to encourage ART adherence. Viral suppression (VS), defined as HIV RNA< 400 copies/ml, was used a marker of ART adherence. Nineteen participating sites in the Bronx, NY and Washington, DC, randomized to the FI intervention, dispensed 39,359 FI gift cards over 2 years. Methods: Semi-structured interviews were conducted with 75 patients ages 4-72 from 4 sites and 7 site investigators (SI) (mostly clinicians); three focus group discussions were conducted with a total of 2 site staff from 0 sites. Data were analyzed to assess opinions about the program and use of FI for VS. Results: Nearly all patients had a positive opinion of the program: they enjoyed receiving the FI, liked that it offered an incentive for improved health, and thought FI could help some people improve ART adherence. At the same time, many patients felt they and others should be selfmotivated to remain adherent and should not have to be incentivized. A few patients felt FI were unnecessary for themselves as they had already achieved VS. SIs were more likely than staff to report positive patient interactions, increased patient adherence to clinic visits and engagement in care. The majority of SIs liked the ability to reward patients, although some felt FI should have been targeted only to low adherers. Several SIs indicated that they had been opposed to FI for VS at the start of the study but were in favor by the end, primarily due to positive patient interactions as SIs were uncertain about the effect on VS. Staff reported implementation challenges and although some reported positive patient interactions, others disliked when patients felt entitled to the FI. Conclusions: Although staff reported some challenges, the FI program was generally well received by patients, SIs and staff, despite the fact that some disagreed with the concept of FI for VS

256 Posters Posters 23: Retention and Adherence in Trials P23.5 Assessment of the Vaginal Residence Time of Biomarkers of Semen Exposure Andrea R. Thurman, Terry A. Jacot, Johan Melendez 2, Thomas D. Kimble, Margaret Christine Snead 3, Roxanne Jamshidi 2, Angie Wheeless 4, David F. Archer, Jill L. Schwartz, Gustavo F. Doncel, Christine Mauck CONRAD, Eastern Virginia Medical School, Norfolk, VA, United States, 2 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 3 Centers for Disease Control and Prevention, Division of Reproductive Health, National Center for Chronic Disease Prevention and Health Promotion, Atlanta, GA, United States, 4 FHI 360, Research Triangle Park, NC, United States P23.6 Implementation of an Electronic Fingerprinting Data Collection System in Zambia: Technical Challenges Kristin M. Wall,2, William Kilembe 3, Mubiana Inambao 4, Roice Fulton 2, Sarah Anderson 3, Alex Tran 2, David Mark 5, Shawn Sarwar 6, Trisha Finnegan 6, Susan Allen 2 Emory University, Atlanta, GA, United States, 2 Rwanda Zambia HIV Research Group, Atlanta, GA, United States, 3 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 4 Rwanda Zambia HIV Research Group, Ndola, Zambia, 5 International AIDS Vaccine Initiative, New York, NY, United States, 6 Biometrac, Washington, DC, United States Background: Biomarkers of mucosal semen exposure can be used to objectively verify protocol compliance and exposure to pregnancy, infection or HIV in clinical trials. The primary objective of this study was to determine the residence time in the vagina (up to 5 days post semen exposure) of 3 biomarkers of semen exposure: prostate specific antigen (PSA) using quantitative total PSA assay, testis specific protein Y encoded 4 (TSPY4) and the sex determining region (SRY) of the Y chromosome, using standard PCR multiplexed with TSPY4 and quantitative PCR. Methods: Healthy women were randomized to unprotected intercourse (n = 7) versus vaginal inoculation with their male partner s semen in the clinic (n = 6), then further randomized to have vaginal swabs obtained at either 7 or 4 time points after semen exposure, up to 5 days post exposure. Post-exposure vaginal swabs were either obtained at home by the participant or in the clinic by the research nurse. Results: When compared to semen inoculation, unprotected intercourse resulted in significantly higher concentrations of semen biomarkers in the first 7 days post exposure. Sampling frequency did not appear to affect biomarker concentrations. Participant sampling at home and nurse sampling in the clinic resulted in similar biomarker concentrations. PSA and SRY, amplified by multiplex PCR, were markers of recent (6-72 hours) semen exposure. TSPY4 and SRY, amplified by qpcr, were reliably detectable for up to 7 days post exposure. Conclusions: We identified biomarkers of recent and longer term exposure to semen, which can be used to assess protocol compliance in contraceptive and microbicide studies. In the latter, semen exposure may be used as a surrogate of HIV exposure. Having swabs obtained at home by study participants is feasible and frequent sampling does not appear to reduce the concentration of these biomarkers. Background: Patient identification ensures data collection accuracy and enhances patient care, including prevention of primary and secondary HIV transmission. However, linking patients within and between health services is an operational challenge in much of sub-saharan Africa. Anonymous electronic fingerprinting systems are a proposed solution. Methods: The feasibility and acceptability of using an electronic fingerprinting system to follow individuals between HIV prevention services was assessed in four Government of Zambia (GRZ) clinics in Ndola, Zambia. The device consists of a Google Nexus 7 tablet with portable single-finger multi-spectral imaging sensor. Templates of scanned fingerprints are sent to a central server via WiFi or mobile. We also pilot tested device sensitivity and specificity under various conditions with Rwanda Zambia HIV Research Group staff in Lusaka and Ndola. Results: Over 50 GRZ clinic-based system users collected fingerprints from 628 HIV testing, antenatal care, male circumcision, and antiretroviral treatment GRZ clinic clients (refusal rate < 5%). Technical challenges included occasional mobile network interruptions, prepaid airtime overruns, and both false positive and false negative fingerprint matching. The first two issues were resolved by caching data during connection interruptions and switching to postpaid airtime. After initial testing among 25 staff, we found device matching was improved by using both thumb and index fingerprints. The device now has a false fingerprint matching rate of /000 and a false rejection rate of < /0,000. Security questions are included as an additional method to confirm matches (year of birth, gender, and father s first name). Conclusions: The anonymous fingerprinting system was acceptable among patients in public health settings. System updates have significantly reduced error rates. We now plan expand the system to other government clinics and to assess acceptability of the device among female sex workers in Ndola and Lusaka. 256 HIV Research for Prevention 204 HIV R4P

257 Posters 23: Retention and Adherence in Trials Wednesday, 29 October P23.7 Using Mixed-methods to Understanding Trial Adherence to a Polyurethane Tenofovir Disoproxil Fumarate Intravaginal Ring in Lowrisk US Women Dana L. Watnick, Marla J. Keller, Lilia Espinoza, Betsy C. Herold, Laurie J. Bauman Albert Einstein College of Medicine, Bronx, NY, United States P23.8 The Research Registry: A Valuable Strategy for Longitudinal Success in HIV Prevention Research Recruitment Renee Weinman, Jonathan R. Baker, Rita Lisa Labbett 2, Sherri Karas, Sharon Riddler, Ian McGowan 2 University of Pittsburgh, Department of Medicine, Pittsburgh, PA, United States, 2 Magee Womens Research Institute, Pittsburgh, PA, United States Background: Vaginal ring delivery of microbicides may overcome adherence challenges conferred by daily or pericoital drug dosing. Results of previous clinical trials have been tainted due to user non-adherence related to user behavior, relationship context, and acceptability. In a Phase trial of a tenofovir disoproxil fumarate (TDF) intravaginal ring (IVR), we are investigating adherence to understand safety and pharmacokinetic outcomes and ultimately design HIV prevention products that are more acceptable to users. Methods: Participants are randomized to receive a polyurethane TDF or placebo IVR for 4 days (d) of continuous use and are asked to remain abstinent and not to insert anything into the vagina or remove the ring. Quantitative (QT) data are collected at baseline, 5 and 4 d after insertion via self-administered computer interview and qualitative (QL) data are obtained via 2 in-depth interviews during and after ring use. QT and QL interview data were triangulated for the first 0 women who completed the trial. Results: Subjects adhered to all study visits. No subject reported per QT or QL that the ring interfered with daily activities or that they removed the ring. On the QT, only reported inserting a finger to check the ring; the QL showed 2 reported checking on it for curiosity or fear of misplacement, and inserted a sex toy. Only woman was somewhat worried about side effects in the QT yet 4 different women were rated as such in the QL. QL data also showed 9/0 women had concerns prior to ring use: nausea, pain, discomfort, long-term unknown effects and infection. Conclusions: Adherence to the study protocol was high by QT but QL identified additional non-adherence to study instructions, more side effect concerns and more fear of potential side effects prior to use that could impede trial enrollment or eventual product use. We recommend mixed-methods to understand and improve participant adherence behavior and QT adherence measurement. Background: The University of Pittsburgh HIV Prevention Research Registry (PRR) was developed to obtain permission from interested participants (ppt) to contact them about new clinical trials for HIV prevention. Methods: The PRR was established and IRB approved in January 202; paper consents were signed followed by a phone interview. In July 202 an online questionnaire was launched eliminating the need for the phone interview, and in January 203 an online consent form was activated. The URL is distributed online, posted on flyers, using social media, and at community outreach events (i.e., health fairs and LGBTfocused events). Attendees at outreach events are invited to join the PRR using ipads. Consented ppts are ed a link and password to complete the questionnaire. Information is stored in a HIPAA compliant database only accessed by research staff. Recruiters use it to select a defined sub-set of ppts who meet study specific criteria. Results: Between April 202 and March 203, 288 ppts joined the PRR via paper consent forms; from April 203 to March 204, 353 joined via the online application. A 23% increase in enrollment occurred after initiation of the online consent. Of the 353 ppts enrolled online, 36 (0%) were referred by friends, 205 (58%) were consented at community events and 36 (0%) joined via social media, with Craigslist being the main source (58%). A 47% increase in questionnaire completion occurred when ppts enrolled using ipads. Over the past 2 months 43 ppts from the PRR were screened and 35 were enrolled in ongoing prevention trials. Conclusions: The registry is a viable strategy for using online applications to provide recruiters with a pre-existing pool of potential participants to launch recruitment initiatives. Since the inception of the registry we have focused on its enrollment, rather than to protocol-specific recruitment. The result has been we now have a greater pool of potential participants for currently enrolling trials as well as for future protocols

258 Posters Posters 24: T-Cell (CD4 & CD8) Responses P24.0 Regulatory T-cells Represent an Important fraction of HIV-specific T-cells: What Is their Impact on Vaccination? P24.02 Seminal Plasma Modulates Dendritic Cell Function Favoring the Generation of CD25+/ FOXP3+ T-cells Vedran Brezar, Nicolas Ruffin, Mathieu Surenaud, Christine Lacabaratz, Karolina Palucka 2,3, Jacques Banchereau,2,3, Yves Lévy,4, Nabila Seddiki Inserm U 955 Université Paris-Est Créteil / The Vaccine Research Institute, Créteil, France, 2 Ralph M. Steinman Center for Cancer Vaccines, Baylor Institute for Immunology Research, Baylor Research Institute, Dallas, TX, United States, 3 The Jackson Laboratory for Genomic Medicine, Farmington, CT, United States, 4 AP-HP, Hôpital H. Mondor - A. Chenevier, Service d Immunologie Clinique et Maladies Infectieuses, Créteil, France Background: Regulatory T-cells (Tregs) play a dual role in HIV infection. Tregs decrease immune activation but also block anti-hivspecific responses. Their role is important to explore for novel vaccines effectiveness. We have reported that vaccination with four administrations of ex vivo generated dendritic-cells loaded with HIV-lipopeptides (LIPO- 5) in patients on therapy was well tolerated and immunogenic (DALIA trial). Vaccine-elicited responses were associated with improved control of viral replication following antiretroviral interruption (ATI) (CROI 202. PB440). Here we investigated the role of Tregs in immunological and virological parameters. Methods: We employed an assay where co-expression of CD25, CD34, CD39 and FoxP3 delineates antigen-specific Tregs and effectors (Seddiki et al. 204). These subsets were studied using samples collected at entry and at week6, 4 weeks after the last immunization (n=4). Results: Upon in vitro PBMCs stimulation, post-vaccination cells showed higher levels of CD25 + CD34 + LIPO-5-specific cells [median (min-max) % to % at entry and wk6 respectively; P= 0.00]. It inversely correlated with viral replication following ATI (r= -0.76, p= 0.00). Simultaneously, there was a shift in antigen-specific effectors/tregs balance and the %CD25 + CD34 + CD39 + FoxP3 + specific- Tregs inversely correlated with %IFNg-producing cells (r= -0.56, p= 0.009). No such changes were noted in these patients when CMV was used as a control. Moreover, Tregs depletion prior to stimulation with LIPO-5-derived antigens, resulted in an increased IFNg-responses (p< 0.05). Conclusions: This study shows that Tregs represent an important fraction of HIV-specific T cells. Tregs impact the frequency and the capability of effector cells to control viral replication. Thus the ability to measure the inducibility of Tregs and/or to modulate those responses prior to vaccination is important for more efficient strategies in future vaccines. Antonela Merlotti, Maria Julia Ruiz, Fernando Erra Díaz, Ezequiel Dantas, Augusto Varese, Gabriel Duette, Pehuen Pereyra, Ernst Glenda, Federico Remes Lenicov, Jorge Geffner, Juan Sabatté Instituto de Investigaciones Biomédicas en Retrovirus y SIDA, Universidad de Buenos Aires/CONICET, Capital Federal, Argentina Background: Unprotected sexual intercourse is the most common mode of HIV- transmission being semen the most important vector for this infection. Dendritic cells (DCs) are abundantly located on mucosal surfaces and play different roles during HIV infection: promote HIV spread by boosting CD4+ T cell infection and activate the HIV specific adaptive immune response. As semen has been shown to promote immune tolerance in different models, we hypothesize that components present in plasma seminal (SP) might modulate DC function promoting a tolerogenic immune response. To test this, we study the ability of complete SP to modulate DC function and the ability of these cells to induce CD25+/FOXP3+ regulatory T cells. Methods: SP was obtained from healthy donors. Monocyte derived DCs were cultured during 24 hs with SP samples (diluted /00) in the absence or presence of LPS (0ng/ml). DC phenotype was studied by flow cytometry. Cytokine secretion was measured in culture supernatants by ELISA. After SP treatment, DCs were cultured with allogeneic CD4+ T cells and the induction of CD25+/FOXP3+ T cells was analyzed by flow cytometry. Results: We found that SP inhibited IL-2, IL-6, TNF-alpha and IL-, but not IL-0 production by LPS stimulated DCs (percent of inhibition respect to LPS alone: 8.2+/-.07% p< for IL-2, /-2.87% p< 0.00 for IL-6, 69.8+/-7.3% p< 0.00 for TNF-alpha, 37.5+/-3.5% p< 0.05 for IL-, 6.+/-5.6% p=0.6 for IL-0). SP also boosted the ability of LPS stimulated DCs to induce CD25+/FOXP3+ T cells (PS+LPS=20,4% vs LPS alone=6%, p< 0,05). We observed no changes on the expression of HLA-DR, CD80, CD86, CD83, CD40 and CDa on immature or LPS-maturated DCs after incubation with SP. Conclusions: SP modulates DC function, inhibiting the secretion of proinflammatory cytokines and favoring the induction of CD25+/FOXP3+ T cells. In this way, we speculate that SP might modulate the adaptive immune response against sexual transmitted pathogens. 258 HIV Research for Prevention 204 HIV R4P

259 Posters 24: T-Cell (CD4 & CD8) Responses P24.03 Wednesday, 29 October P24.04 In vivo Viral Control in a HLA-B*35:0 Homozygous Individual after the Vaccineinduced Response to a Well-defined, HIV Gagderived HLA-B*35 CTL Epitope Miriam Rosas, Beatriz Mothe,2, Núria Climent 3, Maria C Puertas, Javier Martinez-Picado,4,5, Felipe Garcia 3, Christian Brander,4,5, and the RISVAC03 Trial Investigator Team IrsiCaixa AIDS Research Institute - HIVACAT, Badalona, Spain, 2 University of Vic and Central Catalonia, Barcelona, Spain, 3 Hospital Clinic-HIVACAT, IDIBAPS, Barcelona, Spain, 4 Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain, 5 University of Vic and Central Catalonia, Vic, Spain Evolution of Polyfunctional and Proliferative CD8+ T-cell Responses from Early to Chronic HIV- Infection Meika EI Richmond,2, Sandra A. Kiazyk,2, Lyle R. Mckinnon 3, Charles Wachihi 4, Makubo Kimani 4, Joshua Kimani 4, Francis A. Plummer,4,5, T. Blake Ball,2,4 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 2 Public Health Agency of Canada, National Lab for HIV Immunology, Winnipeg, MB, Canada, 3 University of KwaZulu-Natal, Centre for the Programme of AIDS Research in South Africa, Durban, Kenya, 4 University of Nairobi, Nairobi, Kenya, 5 Public Health Agency of Canada, National Microbiology Lab, Winnipeg, MB, Canada Background: Virus-specific CD8 T cell responses to epitopes restricted by HLA-B*35:0 are generally believed to be ineffective in mediating in vivo control of HIV infection. We report a case of a patient homozygous for HLA-B*35:0 who showed successful viral control after vaccination with MVA-B vaccine combined with a drug to reactivate HIV- replication (disulfiram), and who subsequently underwent an analytical treatment interruption (ATI) Methods: Therapeutic vaccination consisted of 3 intramuscular injections of MVA-B at 0,4,6 weeks and a 4th dose followed by 2 months of disulfiram. cart was discontinued 8 weeks after last vaccination. IFN-γ ELISPOT was used to assess immunogenicity and proviral reservoir was determined over time. Viral rebound dynamics were assessed during ATI Results: The patient had a past history of high viral load set point (362,000 copies/ml) and cart was initiated during chronic infection. After ATI the patient remained with low level viral load < 200 copies/ml for >24 weeks without showing a significant decay in CD4 T-cell counts. At baseline, the patient showed a broad (9 different specificities) and strong (9,89 SFC/0 6 PBMC) T cell response, which increased to 4,470 SFC/0 6 PBMC after three vaccinations. Responses to three novel T-cell epitopes present in the vaccine insert (Nef A*03-QK0, Gag A*03-RK9 and Pol B*35-VY0) were induced upon 3 vaccination. A dominant HLA-B*35:0 restricted response to the Gag-p24 B*35-PY9 epitope (PPIPVGDIY) of 3,330 SFC/0 6 PBMC was detected before ATI. No changes in proviral reservoir or viral expression (mrna) were observed in CD4+ T-cells after disulfiram treatment Conclusions: The expansion of a dominant response towards the HLA-B*35- restricted Gag RY9 epitope could potentially explain the observed viral control on this subject suggesting that certain HLA-B*35:0 restricted responses may have the potential to significantly contribute to viral control, providing important guidance for vaccine immunogen design covering non-beneficial HLA alleles Background: The limited success of HIV vaccine candidates to date highlights our need to better characterize protective cell-mediated immunity (CMI). HIV-infected subjects that experience slower progression to AIDS, provide a valuable model for the study of CMI responses that may be capable of controlling HIV. Previous work has demonstrated that these individuals maintain stronger HIV-specific CD8+ T cell proliferation and polyfunctionality compared to progressing controls. However it is unclear whether these CD8+ T cell characteristics are responsible better disease outcomes or if they are merely a consequence of the individuals high CD4+ T cells and low viral loads as a result of protection by other means. Methods: Here we assessed the evolution of CD8+ polyfunctional and proliferative responses from early to chronic HIV- infection in 26 HIV infected individuals following seroconversion using multiparameter flow cytometry. We hypothesized that CD8+ T cells will become increasingly polyfunctional and proliferative from early to chronic infection and patients that mount and maintain an early polyfunctional and proliferative response will have a better disease outcome. Results: Our data suggests that the polyfunctional and proliferative capacity of CD8+ T cells from early to chronic infection follows a distinct pattern, and further that CD8+ T cell responses vary substantially between individuals in the chronic phase of infection. Case study analysis of individuals CD8+ T responses over time suggests that individuals with moderate polyfunctionality in the early phase of infection go on to maintain healthy CD4+ T cell counts. Conclusions: Our data lends support to the hypothesis that polyfunctional and proliferative CD8+ T cells are the cause of slow HIV disease progression. Identification of whether polyfunctional responses and strong proliferative capacity is the cause or consequence of HIV control will be needed for the comprehensive evaluation of HIV vaccine candidates

260 Posters Posters 24: T-Cell (CD4 & CD8) Responses P24.05 P24.06 Identification of CD8+ T-cell epitopes that associate with distinct functionality, proliferation and polyfunctionality Meika EI Richmond,2, Sandra A. Kiazyk,2, Lyle R. Mckinnon 3, Billy Nyanga 4, Charles Wachihi 4, Makubo Kimani 4, Joshua Kimani 2,4, Francis A. Plummer 2,4,5, T. Blake Ball,2,4 Public Health Agency of Canada, National Lab for HIV Immunology, Winnipeg, MB, Canada, 2 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 3 University of KwaZulu-Natal, Centre for the Programme of AIDS Research in South Africa, Durban, Kenya, 4 University of Nairobi, Nairobi, Kenya, 5 Public Health Agency of Canada, National Microbiology Lab, Winnipeg, MB, Canada Background: Understanding correlates CD8+ T cell protection against HIV infection and progressive disease is essential for informing effective vaccine development, design and evaluation. CD8+ T cell responses with a robust polyfunctional and proliferative component are strongly linked to better disease outcomes. However, the specificity of polyfunctional and proliferative CD8+ T cell responses has not been thoroughly investigated. This study provides a better understanding of the fine specificity of HIV-specific CD8+ T cell responses. Methods: Here we have selected eleven HIV- Clade A p24 epitopes of interest, which were previously identified during our comprehensive fine epitope mapping study, for further characterization. Responses to these epitopes were measured in 83 chronically HIV-infected individuals from a Kenyan female sex worker cohort, using multiple cytokines/ chemokines and proliferation. Results: Responses to eleven epitopes were consistent with our previous findings, and most epitope-specific responses were IFN-g negative (64%) while many responses were polyfunctional (38%). We identified epitopes that preferentially elicited specific cytokines/chemokines (p< 0.00) while other epitopes trend towards eliciting a proliferative response. Additionally our data suggests that particular epitopes are associated with polyfunctionality (p< 0.000). Notably, we identified an epitope (EEKAFSPEV) that associates with MIP-β, polyfunctionality and proliferation, all attributes linked with slower disease progression. Conclusions: Importantly, we show that, at a cohort level, particular epitopes preferentially elicit specific qualities of CD8+ T cell responses in preference to others. This suggests there is potential to identify specific epitopes that elicit protective polyfunctional and/or proliferative CD8+ T cell responses. Such protective epitopes could be incorporated into a vaccine to express distinct CD8+ T cell epitopes and induce a more effective CD8+ T cell response. HIV-Specific CD8+ T-cell Expansion Potential, but Not Cytotoxic Capacity, Is Associated with Reduced Set Point Viral Loads in Ad5/HIV Vaccinees Stephen A. Migueles, Nicole Frahm 2, Sushila A. Toulmin, Elizabeth P. Kelly, Bennett A. Peterson, Sarah A. Johnson, M J. McElrath 2, Mark Connors NIAID, NIH, Bethesda, MD, United States, 2 Fred Hutchinson Cancer Research Center, Seattle, WA, United States Background: HIV-specific CD8+ T-cell cytotoxicity is a robust correlate of control in chronic infection, but low cytotoxicity was induced by Ad5/ HIV vaccines. Although some vaccinees maintain control of HIV, the ability of expansion potential or cytotoxic capacity to predict this control has not been examined in the NIAID HVTN 502 Step Study. Methods: Pre-infection PBMCs of 37 vaccinees were analyzed in a blinded manner. In a novel real-time imaging-based assay, cytotoxic responses were measured as the elimination of HIV-GFP-infected CD4+ T-cell targets by 6-day re-stimulated CD8+ T cells. Responses were also measured in LTNP/EC (n=9) and progressors (n=20). True E/T ratios were derived from measures of IFN-gamma+CD07a+ CD8 + T cells and HIV GFP + targets. Responses were correlated with set point HIV RNA levels. Results: Cytotoxicity by confocal imaging was highly correlated with flow cytometric-based assays (r=0.8, p< 0.00). Cytotoxic responses of Step vaccinees were significantly lower than those of LTNP/EC (27.9% v. 68.3%, respectively; p< 0.00) and progressors (43.9%, p< 0.00). Low cytotoxic capacity was not simply due to lower frequency as the per-cell cytotoxic capacity of Step subjects was significantly lower than that of LTNP/EC (p< 0.0). Although the two subjects with HIV RNA set points < 400 copies/ml had high responses, overall cytotoxic capacity did not correlate with viral loads (r=-0.24, p>0.05). However, expansion potential, estimated by Day 6 IFN-gamma+CD07a+ cell frequencies, was weakly correlated with HIV RNA levels (r=-0.42, p=0.0). Conclusions: Pre-infection HIV-specific CD8+ T-cell cytotoxic capacity of Ad5/HIV vaccine recipients in Step was low and did not predict set point HIV RNA levels. A better understanding of the reasons that Ad5/ HIV induces such low levels of per-cell cytotoxic capacity may provide critical insights for induction of an efficacious CD8 + T-cell response by HIV/AIDS vaccines. 260 HIV Research for Prevention 204 HIV R4P

261 Posters 24: T-Cell (CD4 & CD8) Responses P24.07 Wednesday, 29 October P24.08 Pre-exposure Prophylaxis Does Not Enhance HIV-specific T-cell Responses Laura Pattacini, Pamela M. Murnane 2, Jared M. Baeten 3, Tayler R. Fluharty, Katherine K. Thomas 3, Elizabeth Bukusi 4, Elly Katabira 5, Nelly Mugo 6, Deborah Donnell 3, Jairam R. Lingappa 3, Connie Celum 3, Mark Marzinke 7, M. Juliana McElrath, Jennifer M. Lund,3, Partners PrEP Study Team Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 University of Washington, Department of Epidemiology, Seattle, WA, United States, 3 University of Washington, Department of Global Health, Seattle, WA, United States, 4 Kenya Medical Research Institute, Centre for Microbiology Research, Nairobi, Kenya, 5 Makerere University, Department of Medicine, Kampala, Uganda, 6 Kenya Medical Research Institute, Centre for Clinical Research, Nairobi, Kenya, 7 Johns Hopkins University School of Medicine, Department of Pathology, Baltimore, MD, United States Circulating CD4+ T-cells Are Transiently Activated Following Sigmoidoscopy or Circumcision in MSM at High Risk of HIV Infection Maria P. Lemos, Javier R. Lama 2, Shelly Karuna 3, Stephen C. De Rosa 3, Shannon P. Grant 3, Edith M. Swann 4, Carmela Ganoza 2, Silvia M. Montano 5, Jorge Sanchez 2, Juliana McElrath 3,6 Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 Asociacion Civil Impacta Salud y Educacion, Lima, Peru, 3 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 4 NIAID/DAIDS, Bethesda, MD, United States, 5 US Naval Medical Research Unit No. 6, Lima, Peru, 6 University of Washington, Departments of Medicine and Laboratory Medicine, Seattle, WA, United States Background: Pre-exposure prophylaxis (PrEP) is an effective HIV prevention tool in high-risk populations, including subjects participating in vaccine trials. While the primary mode of action for the protective effect of PrEP against HIV infection is likely direct anti-viral activity, nonhuman primate studies suggest that PrEP allows for development of HIV-specific immune responses by aborting infections in HIV-exposed subjects, thus providing a source of immunologic priming. Methods: Within a randomized, placebo-controlled trial among African heterosexual HIV serodiscordant couples in which PrEP demonstrated high efficacy for HIV prevention, we assessed whether enhanced development of HIV-specific T-cell responses was associated with PrEP use. Peripheral blood mononuclear cells from the Partners PrEP Study were used to test for HIV-specific T-cell responses in 230 HIV exposed seronegative (HESN; 79 women, 5 men), half of whom received daily emtricitabine/tenofovir PrEP and half placebo for 2 months; all HESN were randomly selected amongst those with high HIV risk based on an empiric score. HIV-specific CD4+ and CD8+ T-cell responses were detected by intracellular cytokine staining using global potential T-cell epitope mer peptide pools for Gag, Env, and Tat using published protocols. Results: We detected CD4+ T-cell responses to HIV in 8.7% of HESN individuals on PrEP and 9.6% of HESN individuals on placebo (p=0.62). HIV-specific CD8+ T-cell responses were detected in 20.0% of HESN individuals on PrEP and 7.4% of HESN individuals on placebo (p=0.53). The magnitude of the CD4+ and CD8+ T-cell responses did not differ significantly between individuals receiving PrEP versus placebo. Conclusions: In a randomized, placebo comparison, we found no evidence that PrEP usage alters either the frequency or magnitude of peripheral blood CD4 and CD8 HIV-specific responses in HESN. Therefore, it is unlikely that combining PrEP with an HIV vaccine would enhance T-cell immunity. Background: Rectal and genital sampling in vaccine studies in humans permits assessments at the site of HIV entry, yet the impact of circumcision and sigmoidoscopy procedures on peripheral activation readouts is unknown. Methods: Twenty-nine HIV seronegative uncircumcised MSM at high risk of HIV agreed to undergo elective sigmoidoscopy and circumcision in a 28 week cohort study in Lima, Peru, designed to represent mucosal collections in a vaccine study. We monitored peripheral activation before and after the procedures by measuring CCR5, α4β7 and Ki-67/Bcl2 phenotypes among CD4+ T cells by flow cytometry, LPS binding protein (LBP) via ELISA as a marker for bacterial translocation, and cytokine/ chemokine responses using multiplex bead arrays. Results: Compared to immediately prior to sigmoidoscopy, a 2% increase in CCR5+CD4+ T cells (p< 0.002) and a 5.4% in α4β7+cd4+ T cells (p< 0.003) were detected one week post-procedure, and returned to baseline by two weeks post-sigmoidoscopy. Compared to immediately prior to circumcision, an 8.3% increase in CCR5+CD4+ T cells (p< 0.069) and a 9.3% α4β7+cd4+ T cells (p< 0.00) were detected one week post-procedure. A 6.6% increase in CCR5+CD4+ T cells (p< 0.00) and a 5.6% increase in α4β7+cd4+ T cells (p=0.004) were still detected six weeks after circumcision. LBP was significantly increased at one week postcircumcision (8.2 µg/ml) compared to just before the procedure (4.4 µg/ml), but no differences were observed after the sigmoidoscopies or by the end of the six week healing period post-circumcision. No changes in peripheral plasma cytokines were detected throughout the study, suggesting these changes in T-cell activation might reflect localized inflammation at the procedure site. Conclusions: Although the clinical implications of the increase in peripheral immune activation markers following the procedures are unknown, they reinforce the need to provide ongoing risk reduction counseling and support for abstinence recommendations during postprocedure healing. 26

262 Posters Posters 24: T-Cell (CD4 & CD8) Responses P24.09 P24.0 External Quality Assurance Elispot Assay Proficiency Testing in HIV- Clinical Trials in Kenya Robert Kipyegon Langat, Jackton Indangasi 2, Simon Ogola 2, Bashir Farah 2, Peter Hayes 3, Josephine Cox 4, Jill Gilmour 4, Omu Anzala 2 KAVI-Institute of Clinical Research, University of Nairobi, Nairobi, Kenya, 2 KAVI-Institute of Clinical Research,University of Nairobi, Nairobi, Kenya, 3 International AIDS Vaccine Initiative (IAVI)-HIL, Imperial College-London, London, United Kingdom, 4 International AIDS Vaccine Initiative (IAVI)-New York, New York, NY, United States Background: The ELIspot assay has been used both for answering basic research questions and for immunogenicity assessments of HIV- vaccine candidates in clinical trials. As part of the quality management systems, IAVI-sponsored vaccine trial laboratories participate in monthly external quality assurance (EQA) proficiency panels that assess and monitor laboratory performance in ELIspot over time Methods: Frozen PBMC samples isolated from HIV- seronegative individuals with previously-characterized IFN-γ ELISpot responses to CMVpp65, FEC (Flu, EBV and CMV) and mock peptide pools were provided by IAVI from blood packs obtained from the South African National Blood Transfusion Service. Sufficient vials were provided to test the same 6 PBMC samples per month for 6 months. Two such PBMC panels were provided each year. Monthly testing was rotated amongst three laboratory staff at KAVI-Institute of Clinical Research (ICR). Cell viabilities and recoveries were also analyzed Results: Mock data were less than 50 SFU per million PBMC for all sets of PBMC tested over 2 months. 23 out of a total of 26 data points (98.6%) for mock, FEC and CMVpp65 responses were in the expected ranges for PBMC samples tested over 2 months. Intra-operator analysis showed that there were no statistically significant differences in the mock (p=0.35), FEC (p=0.99) and CMVpp65 (p=0.99) responses for PBMC tested over 2 months. Cell recovery was in the range of x 0 6 /ml with viability of above 92%. Conclusions: Three operators have demonstrated competence in ELIspot testing of multiple batches of frozen PBMC over 2 months. Responses were within the expected ranges for mock, CMVpp65 and FEC among different operators. The Elispot proficiency therefore remains a robust and reproducible tool for the assessment of immunogenicity of HIV- and other vaccine candidates in clinical trials Differences in Immunodominant Patterns and HLA-DRB Restriction Characteristics in HIVspecific CD4 T-cell Responses between Clade B and C Infection Faatima Laher, Srinika Ranasinghe 2, Filippos Porichis 2, Isaiah Davis 2, Orestes Mavrothalassitis 2, Nasreen Ismail, Nikoshia Mewalal, Sannie Mahungela, Bruce D. Walker 2,3, Thumbi Ndung u,2, Zaza M. Ndhlovu,2 HIV Pathogenesis Programme, Doris Duke Medical Research Institute, University of KwaZulu-Natal, Durban, South Africa, 2 Ragon Institute of MGH, MIT and Harvard, Massachusetts General Hospital and Harvard Medical School, Boston, MA, United States, 3 Howard Hughes Medical Institute (HHMI), Chevy Chase, MD, United States Background: Increasing evidence suggests that virus-specific CD4 T cells contribute to immune-mediated control of HIV infection. However, the specificities of CD4 T cell responses are poorly defined, particularly in clade C infection. We conducted a comprehensive analysis of virusspecific CD4 T cell responses, defined HLA class II-restriction of detectable responses in clade C infection and compared immunodominant patterns between clade B and clade C CD4 T cell responses. Methods: Immunodominant hierarchies of HIV-specific CD4 T cell responses were defined using CD8 depleted PBMCs in the IFNγ ELISPOT assay. Host genetic effects of class II HLA-DRB alleles on HIV viremia were assessed using the HLA-DRB restriction assay. Immunodominant patterns and restriction characteristics of HIV-specific CD4 T cell responses from our clade C study was compared to previously published data on CD4 T cell responses in clade B HIV infection. Results: Notable differences were identified between the immunodominance hierarchies of HIV-specific CD4 T cell responses in chronic clade C and B infection. In particular, OLP 25 (46-62) in the Gag p24 region, one of the dominant epitopes targeted by 33% of responders in clade C infection was not targeted in clade B infection. The OLP 4 (64-8) epitope, associated with spontaneous control in clade B was dominant in both clade B and C. Consistent with what has been observed in clade B infection, HLA class II DRB restriction in clade C HIV infected individuals showed epitope promiscuity. For example, OLP 63 (76-93) was restricted by both HLA-DRB*3:0 and HLA-DRB*:02. Conclusions: Observed differences in epitope targeting, by HIV-specific CD4 T cell responses, between HIV clade B and C suggests that future vaccines should incorporate immunodominant epitopes targeted by CD4 T cell responses in different clades and ethnicities. Additionally, epitope promiscuity among class II HLA molecules should be taken into account for vaccines designed to induce CD4 T cell responses. 262 HIV Research for Prevention 204 HIV R4P

263 Posters 24: T-Cell (CD4 & CD8) Responses P24. Wednesday, 29 October P24.2 TRIM5α Improves CD8+ T-cell Antiviral Activity and Synergize Intrinsic Restriction and Adaptive Immunity in HIV- Infected Cells Esther Jimenez, Henrik Kloverpis 2,3,4, Ruth Peña, Nuria Izquierdo- Useros, Clotet Bonaventura, Philip Goulder 3, Greg Towers 5, Julia G. Prado AIDS Research Institute IrsiCaixa, Barcelona, Spain, 2 KwaZulu-Natal Research Institute for Tuberculosis and HIV, Durban, South Africa, 3 Peter Medawar Building for Pathogen Research, University of Oxford, Oxford, United Kingdom, 4 University of Copenhagen, Copenhagen, Denmark, 5 University College London, Division of Infection and Immunity, London, United Kingdom Background: TRIM5α restricts HIV- replication in a species-specific manner. Its unusual potency is based on association with the incoming viral capsid inducing proteasome dependent premature uncoating. The role of adaptive immunity in restriction by TRIM5α remains uncharacterised, but is crucial to understanding anti-hiv- protective immunity. Here, we aim to investigate how TRIM5α might contribute to adaptive immunity against HIV-. Methods: Control U937-HLA-B*27:05 cell lines (E) and stable U937- HLA-B*27:05 expressing rhesus TRIM5α (TRIM5rh) or owl monkey TRIM (TRIMCyp) were generated by transduction. Kinetics of HIV- uptake and restriction were measured in all cell lines after infection with HIV-/VSV, encoding GFP. HIV- uptake was measured by p24 staining and virus restriction by enumerating GFP+ cells. To evaluate the effect of TRIM5α on CD8+ T-cell antiviral activity E, TRIM5rh or TRIMCyp expressing cells were infected with HIV-/VSV and co-cultured with HLA-B*27:05 HIV- specific CD8+ T-cells. Relative HIV- CD8+ T-cell antiviral activity was quantified, by enumerating GFP+ cells, and killing by live/death expression in co-cultures. Results: These experiments revealed accumulation of p24 in TRIM5rh or TRIMCyp cells as compared to control cells (E) (7,3%; 3% and 8,7% respectively). In addition, p24 levels inversely correlated with GFP positivity (Spearman, p=0.026), indicating an association between HIV- recruitment and TRIM5α restriction. Our data also show how TRIM5α can significantly increased CD8+ T-cell antiviral activity in TRIM5rh and TRIMCyp infected cells (E vs. TRIM5rh, p=0.0003; E vs. TRIMCyp, p< 0.00). This observation was reinforced by an augmentation of CD8+ mediated T-cell death in TRIM5rh and TRIMCyp infected cells. Conclusions: These data reveal a novel role of TRIM5α acting not only as a restriction factor, but also as an adaptive mediator improving HIV- CD8+ T-cell responses. Long Term Non-progressors from Kigali, Rwanda Display Broad Anti-Gag CD8+ T Cell Responses Nasreen Ismail, Emmanuel Tekirya 2, Susan Allen 2,3, Mark A. Brockman 4, Eric Hunter 2,3, Thumbi Ndung u,5, Etienne Karita 2 Nelson R Mandela School of Medicine, UKZN, HIV Pathogenesis Programme, Durban, South Africa, 2 Project San Francisco, Kigali, Rwanda, 3 Emory University, Atlanta, GA, United States, 4 Simon Fraser University, Vancouver, BC, Canada, 5 KwaZulu-Natal Research Institute for Tuberculosis and HIV, Durban, South Africa Background: The mechanisms of viral control in HIV- infected longterm non-progressors (LTNP) may be informative in vaccine design or other biomedical interventional strategies. Host HLA class I alleles restrict virus-specific CD8+ T cells and have been shown to play a key role in HIV control. In this study, we characterized HLA class I profiles and CD8+ T cell responses in LTNP women from Kigali, Rwanda to explore immunological basis of viral control. Methods: Twenty-one females recruited with HIV- infection in the late 980s in Kigali, Rwanda were longitudinally followed. Nineteen of these women remain therapy naïve, not meeting the national guidelines for combination antiretroviral therapy, and were classified as LTNP. Viral loads, CD4 counts and HLA typing were performed by standard methods. HIV-specific CD8+ T cells were enumerated by IFN-γ ELISPOT assays using PBMC and overlapping consensus clade A peptides spanning the entire proteome. Confirmatory ELISpot assays were performed. Results: The median viral load for this cohort was 4,098 copies/ml (IQR ,05) and the median absolute CD4 count was 498 (IQR ) cells/µl. 42.8% of the women possessed protective HLA alleles (i.e. HLA-B*57/B58:0 and HLA-B*8:0). 95.2% of the women made Gag-specific CD8+ T cell responses. The median magnitude of Gag responses for this cohort was 3,767 spots and the median breadth was 20 peptides. Gag responses dominated the cumulative magnitude at (36.9%), followed by Pol (24.5%), Env (7.8%), Vif/Vpu/Tat/Rev (.6%) and Nef (8.9%). Conclusions: The high frequency of protective HLA class I alleles and high breadth of Gag CD8+ T cell responses in this cohort suggest an important role for host immunogenetics in mediating viral control and LTNP status. Further characterization of immune-mediated mechanisms of viral control in this cohort are warranted

264 Posters Posters 24: T-Cell (CD4 & CD8) Responses P24.3 HIV- Gag and Env Epitope-Specific CD8+ T Cells: Implications for Vaccine Design P24.4 Unbalanced Adaptive and Innate Immune Responses during TB-IRIS Melissa M. Herman, Lyle R. McKinnon 2, Sandra Kiazyk,3, Joshua Kimani 4, Francis A. Plummer,3, T. Blake Ball,3 University of Manitoba, Winnipeg, MB, Canada, 2 CAPRISA, Durban, South Africa, 3 Public Health Agency of Canada, Winnipeg, MB, Canada, 4 University of Nairobi, Nairobi, Kenya Background: Despite the wealth of research being done, an HIV- vaccine has remained elusive, due partly to the extreme genetic diversity of the virus. Stimulating a highly cross-reactive CD8+ T-cell response would help to counter this problem; however, many aspects of the variant-epitope CD8+ T-cell response are still poorly defined. Here, we characterize the CD8+ T-cells specific to common Gag and Env epitopes and their variants, to better understand their function and cross-reactivity. Methods: PBMC samples from HIV+ ART-naive female sex workers from Nairobi, Kenya were collected and stimulated for 6 hours with our antigens of interest from Gag (TL9 epitope variants Ag, M and T) and Env (IF9 epitope variants A, F and L). IL-2, IFNg, TNF and MIPB were measured using intracellular flow cytometry. Each sample was also stained with tetramers specific to the stimulating peptide to assess antigen-specific responses. Proliferation, polyfunctionality and avidity were also measured. Results: This study has revealed that while the A variant was recognized most frequently compared to F and L, there were no differences in cytokine production, polyfunctionality, or proliferation. Conversely, among the three Gag variants, T was recognized by CD8+ T-cells most frequently, and also stimulated the production of more TNF, MIPB and IFNg, and a more polyfunctional, proliferative response. Conclusions: Here, we have shown that the frequency with which an epitope is recognized by CD8+ T-cells does not necessarily determine the strength or function of that response. We have also shown that small mutations within the TL9 Gag epitope can lead to drastically different CD8+ T-cell responses, while changes within the IF9 Env epitope result in few differences. Further, we have seen that the T variant of TL9 produces the best CD8+ T-cell response for combating HIV-. This shows how crucial it is to understand the dynamics of the CD8+ T-cell response to the rapidly mutating HIV- virus, especially when informing vaccine design. Odin Goovaerts,2, Wim Jennes, Marguerite Massinga-Loembé,3, Ann Ceulemans, William Worodria 4,5,6, Harriet Mayanja-Kizza 4,5, Robert Colebunders 7,8, Luc Kestens,2, TB-IRIS Study Group Institute of Tropical Medicine, Biomedical Sciences, Antwerp, Belgium, 2 University of Antwerp, Biomedical Sciences, Antwerp, Belgium, 3 Albert Schweitzer Hospital, Medical Research Unit, Lambaréné, Gabon, 4 Makerere University College of Health Sciences/ Mulago Hospital, Kampala, Uganda, 5 Makerere University College of Health Sciences, Infectious Diseases Institute, Kampala, Uganda, 6 Infectious Diseases Network for Treatment and Research in Africa, Kampala, Uganda, 7 Institute of Tropical Medicine, Clinical Sciences, Antwerp, Belgium, 8 University of Antwerp, Epidemiology and Social Medicine, Antwerp, Belgium Background: Tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) remains a poorly understood complication in HIV-TB patients receiving antiretroviral therapy (ART). TB- IRIS is thought to be associated with an exaggerated immune response to TB antigens. We compared the recovery of IFNγ responses to TBand recall-antigens during TB-IRIS. We also explored the contribution of innate cytokine production to TB-IRIS. Methods: From a prospective cohort study of HIV-TB co-infected patients treated for TB prior to ART initiation, we compared 8 patients who developed TB-IRIS within the first month after ART initiation with 8 non-iris patients matched for age, sex and baseline CD4 count. We analyzed IFNγ ELISpot responses to CMV-lysate, influenza antigen A, PPD, ESAT-6, CFP-0 and LPS at pre-art and IRIS event. CMV, PPD and LPS stimulated ELISpot supernatants were subsequently evaluated for production of IL-2p70, IL-6, TNFα and IL-0 by Luminex. Results: At baseline, before initiation of ART, all measured responses were similar between TB-IRIS group and non-iris controls. During the TB-IRIS event, IFNγ ELISpot responses to TB or influenza antigens were still comparable between TB-IRIS patients and the non-iris patients, but the responses to CMV and LPS were significantly lower in TB-IRIS patients. Production of innate cytokines was similar between IRIS patients and non-iris patients. However, IL-6/IL-0 and TNFα/IL- 0 ratios were increased during TB-IRIS event upon LPS stimulation, compared to non-iris controls. Conclusions: Our TB-IRIS patients did not display an excessive IFNγ response to TB antigens. However, the reconstitution of recall antigen responses is delayed in the TB-IRIS group. In addition, this study reveals an altered innate cytokine balance during TB-IRIS after LPS stimulation. These data provide further arguments for the involvement of the innate immune system in TB-IRIS pathogenesis. 264 HIV Research for Prevention 204 HIV R4P

265 Posters 24: T-Cell (CD4 & CD8) Responses P24.5 Wednesday, 29 October P24.6 Distribution of Bulk and HIV-specific CD8 + T Cell Memory Phenotypes during Acute/Early HIV Infection Is Related to Reduced Antiviral Activity Yanina Ghiglione, Juliana Falivene, Maria Julia Ruiz, Natalia Laufer,2, María Eugenia Socias 3, Pedro Cahn 2,3, Omar Sued 2,3, Horacio Salomón, María Magdalena Gherardi, Gabriela Turk Instituto de Investigaciones Biomédicas en Retrovirus y Sida, (formerly National Reference Center for AIDS), Buenos Aires, Argentina, 2 Hospital Juan A. Fernández, Unidad Enfermedades Infecciosas, Buenos Aires, Argentina, 3 Fundación Huésped, Buenos Aires, Argentina Background: Memory CD8 + T-cells are important components of protective immunity. Understanding their development during primary HIV infection (PHI) may contribute to optimal vaccine design. Aim: To analyze the distribution of memory subsets during PHI and their correlation with functionality and clinical parameters. Methods: 9 samples from acutely infected subjects were obtained at baseline and 2 months post-infection (mpi). Phenotypic (CD45RO, CCR7, PD-) and functional markers (cytokines) were used to identify bulk and HIV-specific CD8 + memory populations. CD8 virus inhibitory assay (VIA) was performed. Data was compared intra-group and correlated to clinical parameters, PD- analysis and CD8 antiviral activity, using nonparametric statistics. Results: Bulk and HIV-specific CD8 + profile was terminal effectors (T E )>naïve >effector memory (T EM )>central memory. Spearman s correlation showed that baseline CD8 + VIA inversely correlated with the concurrent proportion of HIV-specific CD8 + T EM cells (r=-0.593, p=0.009) and directly correlated with the proportion of HIV-specific CD8 + T E cells (r=0.78, p=0.0008). Identical correlations were observed between baseline CD8 + T cell phenotype and CD8 + VIA at 2 mpi. Also, percentage of PD- high CD8 + T cells negatively correlated with bulk and HIV-specific CD8 + T EM cells (r=-0.50, p=0.034 and r=-0.668, p=0.004, respectively). Conversely, positive correlations were observed with the proportion of bulk and HIV-specific CD8 + T E cells (r=-0.50, p= and r=-0.564, p=0.022, respectively). Conclusions: A higher proportion of fully differentiated HIV-specific cells are related to the magnitude of CD8 + antiviral activity (rapidly able to exert effector functions) and to a higher PD- expression (related to T cell differentiation stage and activation status). This is the first report were a relation between CD8 + T cell memory differentiation hierarchy and antiviral function is reported during acute infection, providing information potentially useful for vaccine design. T-cell Responses Targeting HIV Env V2 in Natural Infection: Implications for RV44 Vaccine Recipients Andrew Gartland, John McNevin, Pradyot Dash 2, Paul Thomas 2, Tomer Hertz, Julie McElrath Fred Hutchinson Cancer Research Center, Vaccines and Infectious Disease, Seattle, WA, United States, 2 St. Jude Children s Research Hospital, Immunology, Memphis, TN, United States Background: The RV44 HIV- vaccine trial showed 3% efficacy, with Env V/V2-specific antibodies inversely correlated with infection risk and viral genetic signatures of immune selection in the V/V2 of breakthrough viruses. A sieve analysis to detect vaccine-induced, T-cell mediated viral escape showed that the selection could be partly attributed to T cells and that an HLA allele may have modified vaccine efficacy. Studies suggest vaccine-induced CD4+ T-cells preferentially targeted V2. Due to the high genetic variability, responses to V2 have not been adequately assessed in vaccine recipients, nor in HIV infection. Accurate quantification of V2-specific T-cell breadth and depth requires a unique peptide library. Methods: We devised an algorithm to design peptide libraries for mapping T cell epitopes in variable regions. Optimal peptides are selected for HLA binding to a frequency-weighted set of alleles and for sequence diversity coverage. We applied this algorithm, focusing on subtype B variants of one epitope implicated in the sieve analysis KMQKEYALL (KL9, HXB ). We used IFNγ ELISpot, multi-panel ICS and single-cell TCR sequencing to characterize KL9-specific T-cells in chronically infected individuals and LTNPs. Results: Robust, cross-reactive T-cells were identified, some responding to 5 KL9 variants. Strikingly, one LTNP had avid, polyfunctional, A*24- restricted CD8+ T-cells recognizing 6 KL9 variants including the subtypes CRF0_AE and B in the RV44 vaccine and it was restricted by a single T cell clonotype. The data suggest that despite sequence variability, T-cell responses to this region may recognize most circulating variants. Conclusions: T-cell responses to variable regions are more effectively mapped using optimized peptide libraries. The V2-region of Env is a T-cell epitope hotspot requiring further characterization in natural infection and vaccine trials. This motivates further studies of the role vaccine-induced V2-specific T-cell responses played a in the RV44 trial efficacy

266 Posters Posters 24: T-Cell (CD4 & CD8) Responses P24.7 HIV--specific Cross-reactive CD8 T Cells Demonstrate Compromised Killing Capacity during Acute Infection P24.8 A Distinct Treg Transcriptome Signature in HIV- Elite Controllers Might Contribute to Improved Disease Outcome Victor Yimin Du, Anju Bansal, Jonathan Carlson 2, Jesus Salazar- Gonzalez, Sonya L. Heath, Eric Hunter 3, Paul A. Goepfert University of Alabama at Birmingham, Medicine, Birmingham, AL, United States, 2 Microsoft Research, Los Angeles, CA, United States, 3 Emory University, Pathology, Atlanta, GA, United States Background: Human immunodeficiency virus (HIV-) escapes from CD8 T-cell responses by adapting to host human leukocyte antigen class I (HLA-I) alleles, forming adapted epitopes (AE) that can be transmitted to new hosts. Prior studies on the first antiviral CD8 T-cell recognition of AE in HIV- clade B acutely infected patients revealed that AE-specific primary T-cell responses existed in rarer frequencies and exhibited poorer killing capacity, as compared to their non-adapted (NAE) counterparts. We would like to determine the ability of NAE-specific T cells to crossrecognize AE, as plasticity in antigen targeting has been anticipated for viral-specific CD8 T cells. Methods: We gathered acute PBMC samples that contain CD8 T cells previously found to respond against NAE in autologous founder virus, and stimulated them with common AE variant peptides in an IFNg ELISPOT assay to test for cross-reactivitiy. Results: Out of all pre-existing primary NAE-specific T-cell responses, there was a trend towards fewer number of NAE that demonstrated AE cross recognition compared to the ones that did not (28.57% vs. 7.43% respectively, p=0.06). While NAE-specific T cells efficiently killed target cells that were pulsed with the said NAE peptide, the corresponding cross-reactive killing of cells pulsed with AE variants was poorly induced. Cross-reactive CD8 T-cell responses also exhibited lower antigen sensitivity compared to the counterpart primary T-cell responses, suggesting that avidity in T-cell recognition may drive difference in killing capacity between the two types of response. Conclusions: Overall, our study suggests compromised T-cell cross recognition of AE during acute HIV infection, which complements prior studies demonstrating that CD8 T-cell mediated HIV adaptation impairs early viral control. Christine Dahlke, Mathieu Angin 2,3, Christian Müller 4, Siddharta Sharma 2, Manoj K. Bhasin 5, Yan Zhuang 2, Marylyn M. Addo,6,7 Universitätsklinikum Hamburg-Eppendorf, Department of Medicine, Emerging Infections, Hamburg, Germany, 2 Ragon Institute of MIT, MGH and Harvard, Boston, MA, United States, 3 Institut Pasteur, Virology, Paris, France, 4 University Heart Center Hamburg, Clinic for General and Interventional Cardiology, Hamburg, Germany, 5 Beth Israel Deaconess Medical Center, Genomics & Proteomics Core, Boston, MA, United States, 6 German Center for Infection Research, Emerging Infections, Hamburg, Germany, 7 Massachusetts General Hospital, Division of Infectious Diseases, Boston, MA, United States Background: Regulatory T cells (Tregs) suppress immune system activation and promote immunologic tolerance. Due to their impact on both innate and adaptive immune responses, Tregs have recently been explored as new targets for immunotherapy and vaccination strategies, as in cancer and transplantation settings. Moreover, Tregs garnered considerable interest in HIV pathogenesis, due to their potential impact in HIV-associated immune activation and viral replication. However, the role of Tregs in HIV infection remains incompletely understood. Here, we performed the first large scale transcriptome analysis of Tregs in HIV-infected individuals, which may provide important insight into Treg biology and support strategies for immune-based therapies and HIVvaccine design. Methods: We investigated the gene expression profiles of Tregs and conventional CD4 + T cells (Tconv) in HIV elite controllers (EC), individuals with chronic untreated HIV- infection (CU) and HIV-negative individuals. Following flow-based cell-sorting of Tregs and Tconv, whole genome expression analysis was performed using Illumina BeadChip technology. We screened for differential gene expression between the two T cell subsets and the study cohorts using the software R/bioconductor packages lumi and limma. Interesting candidate genes were further validated and investigated in ex vivo studies. Results: We identified distinct group-specific variations in Treg signatures between EC and CU - with more than 40 genes significantly up/downregulated. Those include several genes associated with interferon response (eg. IFI44) or transcriptional regulation (eg. PBX2), which are potentially relevant for the outcome of viral infections. Additionally, we observed HIV- and Treg-specific regulated genes in line with previously published studies, validating our experimental setup. Conclusions: Our findings provide novel insight into the impact of Tregs on HIV pathogenesis. The results might be fundamental for novel concepts in HIV immunotherapy and vaccine design. 266 HIV Research for Prevention 204 HIV R4P

267 Posters 24: T-Cell (CD4 & CD8) Responses P24.9 Wednesday, 29 October P24.20 LB HIV Minor Variants Detected by Next Generation Sequencing: Impact on Immune Control of HIV in the Context of HLA-B*27:05 and HLA-B*57:0 Jacqui Brener, Astrid Gall 2, Rebecca Batorsky 3, Paul Kellam 2, Todd Allen 3, Philippa Matthews, Philip Goulder Peter Medawar Building for Pathogen Research, University of Oxford, Oxford, United Kingdom, 2 Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 3 Ragon Institute of MGH, MIT and Harvard, Boston, MA, United States HAART Alters Multiple Cellular Peptidase Activities and Predictably Modifies Protein Degradation Patterns, Epitope Production and Presentation Georgio Kourjian, Hugo Poplimont, Ijah Mondesire-Crump, Matthew Berberich, Hang Su, Sylvie Le Gall Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States Background: The advent of next generation sequencing technology has provided a sensitive platform for producing extensive datasets that allow novel and detailed analyses of viral adaption and immune control of HIV. In the context of expression of HLA alleles associated with superior viral control and slow disease progression, in particular HLA-B*27:05 and HLA-B*57:0, intrahost population diversity revealed through the minor variant data produced may provide critical insights into the determinants of immune control. Methods: We performed full-length HIV high-throughput Illumina sequencing on a cohort of subjects expressing both HLA-B*27:05 and HLA-B*57:0, including two transmission pairs and eleven additional individuals, with longitudinal sampling of six subjects. The HLA- B*27:05/B*57:0 positive group included both HIV controllers and progressors. Results: In order to evaluate the determinants of HLA-mediated control of HIV, we have conducted an analysis of the minor variant populations and their impact on immune control, comparing intrahost variability across the full genome and at CD8+ T Cell epitopes in HLA-B*27:05/B*57:0 positive controllers versus non-controllers. We describe changes in intrahost variability over time, and analyse the evolution of the donor founder virus in the context of HLA-B*27:05/B*57:0 expression in the transmission pairs. In an HLA-B*27:05/B*57:0 positive progressor we show that accumulation of escape mutations at low levels in the minor variants precedes fixation of these mutations within the population and predicts disease progression. Conclusions: The novel insights afforded through use of next generation sequencing technology may be critical for furthering our understanding of intrahost viral evolution and immune control. Particularly in the context of expression of favourable HLA alleles, these insights may provide important information for informing HIV T cell vaccine design. Background: Epitopes displayed by MHC-I and MHC-II result from the intracellular degradation of antigens by multiple proteases and peptidases located in various subcellular compartments. We and others previously showed that due to structural homologies HIV proteaseinhibitors (PIs) used in antiretroviral therapies (ART) affect proteasome activities in PBMC. Addition peptidases and several intracellular processing pathways are required for the complete production of MHC-I and MHC-II epitopes but the effect of ART on these pathways is not known. Methods: Eight cellular peptidase activities were assessed in PBMCs, CD4 T cells, dendritic cells and macrophages treated with seven HIV PIs, NRTIs or NNRTIs. The impact on epitope presentation was measured by T cell killing assay. Using in vitro degradation of proteins in subcellular compartments of primary cells and mass spectrometry analysis we followed protein degradation into epitopes. Through computational analysis of degradation products we identified alterations in cleavage sites of antigenic proteins. Results: HIV PI-induced alterations of cellular peptidase activities were variable according to PI, cell types and peptidases. PIs altered different steps of antigen processing: degradation patterns, amount and intracellular stability of epitopes. They affected MHC-I and MHC- II epitope production. Altered degradation patterns led to increased or decreased frequency of cleavage sites observed in control conditions and appearance of new cleavage sites, leading to the production of putative new epitopes and modified production of known epitopes. Accordingly HIV PIs increased or decreased recognition of HIV-infected cells by CD8 T cells. Conclusions: Antiretroviral therapies including HIV PIs alter cellular peptidase functions in a sequence-dependent manner. This may lead to a diversification of pathogen-derived epitopes and possibly of immune responses against residual HIV or co-infecting pathogens in HIV-infected persons

268 Posters Posters 25: Transmission and Viral Diversity P25.0 Biomarkers of Cervical Inflammation and Immunity Associated with Cervical HIV- RNA Christine K. Mauck, Pai-Lien Chen 2, Charles Morrison 2, Raina Fichorova 3, Cynthia Kwok 2, Tsungai Chipato 4, Robert Salata 5, Gustavo Doncel CONRAD, Arlington, VA, United States, 2 FHI 360, Durham, NC, United States, 3 Brigham and Women s Hospital and Harvard Medical School, Boston, MA, United States, 4 University of Zimbabwe, Harare, Zimbabwe, 5 Case Western Reserve University, Cleveland, OH, United States P25.02 Defining HIV/SIV Particle Localization and Infection after Rectal Challenge in the Rhesus Macaque Model using Reporter Systems Wesley A. Grimm, Luis Barcena, Danijela Maric, Gianguido C. Cianci, Daniel Stieh, Shannon Allen, Michael McRaven, Ron S. Veazey 2, Thomas J. Hope Northwestern University, Chicago, IL, United States, 2 Tulane National Primate Research Center, Covington, LA, United States Background: Cervicovaginal HIV shedding increases HIV transmission. Genital tract inflammation increases shedding through production of cytokines/chemokines which recruit and activate HIV target cells. We evaluated whether cervical immune mediators present before HIV acquisition affected later HIV shedding and whether after acquisition, levels of mediators differed between shedders and non-shedders. Methods: We used cervical samples from 57 women with welldocumented HIV seroconversions during the Hormonal Contraception and HIV (HC-HIV) study in Uganda and Zimbabwe. Samples were from the visit before (T-), at (T0), and 2 visits after seroconversion (T+ and T+2). We measured 8 biomarkers of inflammation and immunity (IL-β, IL-RA, IL-6, IL-8, RANTES, MIP-3α, VEGF, ICAM-) by electrochemiluminescence multiplex and antibacterial proteins SLPI and BD-2 by ELISA. We used the Wilcoxon test and generalized linear models to evaluate the association between mediator levels and shedding, and other covariates impact on shedding. Results: The only immune mediator at T- associated with women who shed (vs. those who did not) at T0 (seroconversion) was RANTES (p=0.05). In both shedders and non-shedders, seroconversion was followed by a significant increase in RANTES and decrease in IL-8 and VEGF (T- vs T0). Among non-shedders only, IL-6 and BD-2 decreased and MIP- 3α and ICAM- increased while, among shedders, SLPI decreased after seroconversion. The magnitude of changes in RANTES, SLPI and MIP- 3α between T- and T0 was significantly greater in shedders vs. non shedders. At T0, shedders had lower levels of SLPI and MIP-3α than non-shedders. The difference remained significant at T+ for MIP-3α. Conclusions: Specific immune mediator profiles are associated with risk of HIV acquisition and cervical shedding. Increased RANTES and decreased SLPI were associated with increased risk of HIV shedding, possibly due to their effects on HIV target cells and innate immunity. Background: Receptive anal intercourse accounts for over half of new HIV- infections in the United States each year; yet, how HIV- virions penetrate rectal mucus and enter the lamina propria to access target cell pools is an understudied area. Methods: HIV- mobility was assessed ex vivo within human rectal mucus by fluorescence microscopy. Imaging was used to assess in vivo penetration frequency and depth of photoactivatable HIV 4 hours after atraumatic intrarectal HIV challenge in macaques. To define the targets of infection after rectal challenge in the rhesus macaque model, we challenged with bicistronic vectors expressing luciferase and fluorescent proteins. 3 days post challenge luciferase was utilized to identify foci of infected cells, fluorescent protein expression was utilized to identify infected cells in cryosections of the luciferase positive tissue. Results: HIV- and nanoparticle mobility within human rectal mucus is very similar to that observed for cervical mucus in women. This mucus forms a formidable barrier to the virus as most virions were found to be trapped in the mucus layer 4 hours after challenge of human explants and living macques. In vivo penetration of single virions occurs in macaque rectal tissues as early as 4 hours post-innoculation, however, penetrating foci containing multiple virions were not readily observed. We detected infected foci at biopsy sites of rectal tissues by luminescence. HIV-enveloped virions enter rectal tissues and infect primarily CD68+ macrophages. Conclusions: Rectal mucus inhibits HIV mobility revealing that mucus acts as an important initial barrier in vivo. Together, this data suggests that atraumatic inoculation may result in diffuse infection of single cells within the rectal tissues in contrast to the focal infection observed after vaginal challenge. A better understanding of the sites and target cells of rectal transmission will inform vaccine and microbicide design. 268 HIV Research for Prevention 204 HIV R4P

269 Posters 25: Transmission and Viral Diversity Wednesday, 29 October P25.03 P25.04 Development of Molecular Technique for Deep Sequencing of almost Full HIV Genome Dmitry E. Kireev, Alexey E. Lopatukhin, Alexey D. Neverov, Gennady G. Fedonin, Andrey B. Shemshura 2, Sergey R. Saukhat 2, German A. Shipulin Central Research Institute for Epidemiology, Department of Molecular Diagnostics and Epidemiology, Moscow, Russian Federation, 2 Rostov Research Institute of Microbiology and Parasitology, Rostov-on-Don, Russian Federation Background: Assays for identification of HIV drug resistance (DR) mutations are becoming expensive and laborious due to growth of quantity of HIV-infected people and number of drugs. Next generation sequencing (NGS) has a significant advantage over conventional population sequencing which is used in routine diagnostic. Implementation of NGS in diagnostic practice will greatly reduce the complexity and the cost of DR testing. Our aim was to develop the molecular method for deep sequencing of all viral genes and compare the results with conventional population sequencing. Methods: The blood plasma from HIV-positive person who has been infected in 990 and changed 5 schemes of therapy at the sampling date was used as a clinical material. SuperScript III RT-PCR System was applied for RT-PCR and in-house reagents for the second round of amplification. Totally there were amplified more than 9000 nucleotides with 5 overlapping fragments. These fragments were sequenced using ABI 3500 and MiSeq in parallel. Data obtained by the population sequencing were analyzed by BioEdit and DEONA software. Analysis of NGS data was done by the in-house algorithm. Results: Following results were obtained from the NGS data: total capacity nucleotides, reads; total length of the consensus sequence nucleotides (98.58% of HXB2); maximum depth coverage reads; average reads; depth less than 000 reads - 89 nucleotides (,97%). The concordance between the consensus sequences of NGS and the population sequence (total length nucleotides) was 99,25%. Thorough analysis revealed that only 48 positions were different and there were found complete mismatches only in 0 positions. NGS consensus contained all DR mutations which were found in population consensus. Conclusions: We developed the molecular method which allows getting near full HIV genome by amplification of overlapping fragments and following NGS. Comparison with population sequencing revealed the high degree of concordance. Increasing Diversity of HIV- Based on Envelope and Pol-RT Sequences of Viruses Circulating among Antiretroviral Experienced Kenyans Rose C. Kitawi,2, Timothy Nzomo,2, Ruth Sada,2, Maureen Kimulwo,2, Geoffrey Masankwa, Bernhards Ogutu,3, Rashid Aman,3,4, Washingtone Ochieng,3,5 Center for Research in Therapeutic Sciences, Nairobi, Kenya, 2 Institute of Tropical Medicine and Infectious Diseases, JKUAT, Nairobi, Kenya, 3 Kenya Medical Research Institute, Nairobi, Kenya, 4 African Centre for Clinical Trials, Nairobi, Kenya, 5 Harvard Medical School, Boston, MA, United States Background: The HIV- virus is highly variable and continuous understanding of viral diversity is important for ongoing therapeutic and preventive strategies. Studies have separately shown that HIV- strains are differentially distributed across different regions in Kenya. The rapid divergence of strains from initially predominant subtype A epidemic poses a challenge in the application of mitigation strategies against the epidemic. We describe the pattern of viral diversity among Kenyans receiving treatment and care in comprehensive care centres. Methods: Patients aged between 3 and 64 years were consented and recruited. Blood samples obtained from each patient were used to prepare plasma and Peripheral Blood Mononuclear Cells (PBMCs). DNA from PBMCs and RNA from plasma were amplified by envelope and pol-rt specific primers and sequenced. Sequences obtained were edited and aligned. Neighbour-joining phylogenetic trees were constructed using Kimura s two-parameter distances. Results: Out of 50 patients with low to undetectable plasma viral load,the distribution of DNA-derived strains based on envelope c2v3 region was 48% for subtype A, % (subtype B), 2% each for subtypes C and D, and 42% mixed or discordant subtypes and circulating recombinant forms. From cell-free plasma of patients with >000 HIV- RNA copies, the genotypes based on 40 Pol-RT sequences were 52.5% pure HIV-A, 2.5% pure HIV-B, 5% HIV-C, 0% pure HIV-D and 22.5% CRF0_ AE and % CRF0_BC recombinants. The cell-free compartment had mainly pure viral genotypes while the cellular compartments had near proportional levels of pure and mixed or discordant strains. Conclusions: We demonstrate a declining dominance of HIV- subtype Ain Kenya, the emergence of previously unreported subtype B and increasing CRF0_AE. These data will prove valuable in developing strategies for public health and therapeutic or preventive interventions

270 Posters Posters 25: Transmission and Viral Diversity P25.05 Semen-mediated Enhancement of HIV Infection Markedly Impairs the Antiviral Efficacy of Microbicides Onofrio Zirafi, Kyeong-Ae Kim, Nadia R. Roan 2, Benjamin Mayer 3, Silvia Kluge, Shibo Jiang 4, Warner C. Greene 2, Frank Kirchhoff, Jan Münch Ulm University Medical Center, Institute of Molecular Virology, Ulm, Germany, 2 University of California San Francisco, CAPS, San Francisco, CA, United States, 3 Ulm University, Ulm, Germany, 4 New York Blood Center, New York, NY, United States P25.06 Serial in vivo Passage of CXCR4-tropic Simian/Human Immunodeficiency Chimeric Virus (SHIV 89.6P ) in Papio Anubis; A Preclinical Disease Model Shem P. M. Mutuiri,2, Gerald Chege 3, Lucy A. Ochola, Elephas W. Munene, Maria Kiio, Peter G. Mwethera, Thomas M. Kariuki Institute of Primate Research (IPR), Tropical Infectious Diseases, Nairobi, Kenya, 2 University of Nairobi, Please Select, Nairobi, Kenya, 3 University of Cape Town, Medical Virology; Clinical Laboratory Sciences, Faculty of Health Sciences, Cape Town, South Africa Background: Topically applied microbicides potently inhibit HIV in vitro but largely failed to exert protective effects in clinical trials. We explored whether the ability of semen to enhance HIV infection affects the antiviral efficacy of various classes of microbicides and antiretrovirals (ARVs). Methods: HIV infection assays were performed in the presence or absence of semen (or SEVI amyloid fibrils) in cell lines and primary PBMCs; IC 50 values of ARVs were determined and evaluated. Results: We demonstrate that the ability of semen to enhance HIV infection in vitro markedly impairs the antiviral efficacy of ARVs that target virion components by 0 to 20-fold. These ARVs include polyanions, neutralizing antibodies, NRTI and NNRTI s, and inhibitors against HIV- Integrase and Protease. Similar results were obtained using synthetic SEVI fibrils. In contrast, semen deficient of amyloids and lacking the ability to enhance HIV infection did not impair the antiviral activity of ARVs. In direct contrast, the CCR5 antagonist Maraviroc (MVC) blocked mock and semen-exposed virus with similar efficacy. Notably, the concentrations of MVC required to block semen-exposed virus infection of PBMCs are lower than those that can be achieved in the genital tract after oral administration of the drug. Conclusions: Our data show that the HIV enhancing activity of amyloids in semen undermines the antiviral efficacy of ARVs that target viral components, which might explain why such microbicides largely failed in clinical trials. In contrast, MVC, which targets a host protein that is present at constant amounts at the cell surface, retained activity in the presence of semen. These results suggest that compounds targeting cellular components may be advantageous for microbicide development. Since semen is the main vector for the spread of the AIDS pandemic, we recommend testing future candidate microbicides against semen-exposed virus to identify those agents that retain potent antiviral activity during sexual virus transmission. Background: SHIV is widely used in HIV research to assess the efficacy of AIDS vaccines and anti-hiv microbicides using the macaque animal model. The objective of this study was to generate a baboon-adapted SHIV 89.6P, to allow similar studies to be conducted in the baboon, which is already a well-characterised model at IPR for reproductive biology, schistosomiasis and malaria. Methods: To ensure pathogenicity and increase the virulence of stock SHIV 89.6P through in vivo serial passage. To achieve this, SHIV 89.6P was serially passaged in vivo in three juvenile baboons via direct intravenous (I.V.) injection of undiluted cell culture supernatant in the st baboon and I.V. inoculation of heparinised whole blood and bone marrow (BM) from inoculated baboon into subsequent baboons Results: Two to five weeks after inoculation all animals were virus culture positive, in CEMX74 and HUT78 cell lines. Viral antigens were readily detected in the day 4 post-inoculation plasma (75, 72 and 2,287 pg/ml for the st, 2 nd & 3 rd passages respectively). Titration of the inoculum in CEMX74 cells yielded higher viral titres, ranging from 0 6 to TCID-50, compared to the original inoculum at TCID-50. Viral Gag (SIVp27) measurements in inocula confirmed that the baboongenerated SHIV inoculum was several-fold higher (>0 mg/ml) than the initial virus stock (0.03 mg/ml). Conclusions: These observations indicate that in vivo serial passaging in baboons successfully generated a baboon-adapted SHIV 89.6P. This can be utilized for future studies using the pre-clinical model which is anatomically, immunologically and genetically closer to humans and vulnerable to various human diseases facilitating disease, co-infection and vaccine studies. 270 HIV Research for Prevention 204 HIV R4P

271 Posters 25: Transmission and Viral Diversity Wednesday, 29 October P25.07 HIV-enhancing Amyloids Are Prevalent in Fresh Semen and Are a Determinant for Semen s Ability to Enhance HIV Infection: Relevance for HIV Transmission Shariq Usmani, Haichuan Liu 2,3, Chris D. Pilcher 4, H. Ewa Witkowska 2,3, Frank Kirchhoff, Warner C. Greene 5,6, Jan Münch, Nadia R. Roan 5,7 Ulm University Medical Center, Institute of Molecular Virology, Ulm, Germany, 2 UCSF, Department of Obstetrics, Gynecology & Reproductive Sciences, San Francisco, CA, United States, 3 UCSF, Sandler-Moore Mass Spectrometry Core Facility, San Francisco, CA, United States, 4 UCSF, HIV/AIDS Division, San Francisco General Hospital, San Francisco, CA, United States, 5 The J. David Gladstone Institutes, Gladstone Institute of Virology and Immunology, San Francisco, CA, United States, 6 UCSF, Departments of Medicine and Microbiology and Immunology, San Francisco, CA, United States, 7 UCSF, Department of Urology, San Francisco, CA, United States P25.08 Metagenomics Analysis of Plasma in HIVinfected Men Who Have Sex with Men in Bangkok, Thailand Wiriya Rutvisuttinunt, Wanna Leelawiwat 2, Piyawan Chinnawirotpisan, Famui Mueanpai 2, Oranuch Kongpechsatit 2, Chonticha Klungthong, Robert O Connell 3, Mark de Souza 4, In- Kyu Yoon, Marcel Curlin 2,5, Stefan Fernandez Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 2 The Thai Ministry of Public Health U.S. Centers for Disease Prevention Collaboration, Nonthaburi, Thailand, 3 Department of Retrovirology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 4 South-East Asian Research Collaboration with Hawaii, Thai Red Cross AIDS Research Center, Bangkok, Thailand, 5 Division of HIV/AIDS Prevention, Centers for Disease Control and Prevention, Atlanta, GA, United States Background: Semen, the most common vehicle for HIV transmission, enhances HIV infection in vitro. Previously, naturally-occurring peptides derived from the semen proteins prostatic acid phosphatase (PAP) and semenogelin (SEM) were shown to polymerize into amyloid fibrils that markedly enhance HIV infection. Here, we investigated whether endogenous amyloid fibrils can be detected in fresh semen, and if so the extent to which they contribute towards the ability of semen to enhance HIV infection. Methods: Confocal microscopy, electron microscopy, atomic force microscopy, quantitative mass spectrometry, ELISAs, and viral infection assays were used to detect, quantitate, and characterize endogenous HIV-enhancing amyloids in semen. Results: Endogenous PAP and SEM amyloids were directly visualized in unmanipulated semen by immunogold electron microscopy and confocal microscopy. Amyloid structures were further characterized by atomic force microscopy. We additionally demonstrated that endogenous amyloids are present in semen from men acutely infected with HIV, and that these fibrils directly bind HIV virions. We then examined whether the extent to which semen enhances HIV infection correlates with the levels of endogenous amyloids. We found that the magnitude of HIVenhancing activity of semen is dependent on the semen donor and the degree of liquefaction, and that the levels of the HIV-enhancing PAP and SEM amyloidogenic fragments significantly correlate with the variability in enhancing activity observed between samples. Accordingly, semen completely deficient in amyloids, due to a condition termed ejaculatory duct obstruction (EDO), lack the ability to enhance HIV infection. Conclusions: Our results demonstrate that endogenous semen amyloids are a crucial determinant for semen-mediated enhancement of HIV infection, and emphasize the need to consider the effects of semen in approaches to minimize the spread of HIV. Background: Men who have sex with men (MSM) are considered a highrisk group contributing to global HIV- persistence despite falling HIV- incidence rates in other groups. Understanding microenvironments and potential co-infecting pathogens offer the possibility of developing novel and effective measures for prevention and treatment of HIV in this group. We determined the presence of viral metagenomes and the relationship with HIV- infection in samples collected from Thai MSM. Methods: We selected 7 HIV--infected plasma samples obtained from untreated MSM with chronic or recently acquired HIV- infection collected between July 2006 and March 200. Sample preparation was performed using NextaraXT without any specific genome amplification steps, followed by sequencing on MiSeq Illumina. We conducted metagenomic data analysis by de novo assembly and read-mapping alignment. Results: A total of 0.96 million genomic DNA sequences(mean 323,544; range 65,028-,437,03 reads) was obtained from all specimens. Thirteen of 7 specimens had detectable reads from infecting pathogens sufficient for further analysis. Eleven of 3 specimens contained a single predominant viral pathogen; GB Virus C (GBV-C) was present in 5/ specimens, HIV- was detected in 4/ and hepatitis B virus (HBV) was found in 2/. Two of 3 specimens had multiple pathogens: one contained GBV-C and hepatitis C virus (HCV) and the other contained GBV-C and HBV. Six out of seven specimens containing GBV-C were collected from chronically HIV- -infected individuals. Conclusions: Co-circulating viral pathogens may be common in some subgroups of HIV--infected MSM. In this study, GBV-C was most commonly noted in chronic rather than acute infection. The low rate of HIV- detected reflects the lower abundance of HIV- genomes relative to genes from the host and co-infecting pathogens. The significance of co-infecting pathogens in HIV- infection is not known but may affect the course of disease progression. 27

272 Posters Posters 25: Transmission and Viral Diversity P25.09 Characterization of Putative Ancient HIV- Lineage Viruses: Insight into the Early Epidemic of HIV- Marcel Tongo,2, Wendy A. Burgers 3, Melissa-Rose Abrahams 3, Darren P. Martin 3, Jeffrey Dorfman International Centre for Genetic Engineering and Biotechnology, University of Cape Town, Cape Town, South Africa, 2 Institute of Medical Research and Study of Medicinal Plant (IMPM), Yaounde, Cameroon, 3 IDM, University of Cape Town, Cape Town, South Africa P25.0 Drug Resistant SIV Strains Carrying Higher-Fidelity K65R and Q5N Reverse Transcriptase Mutations Are Less Fit in vivo Sarah B. Lloyd, Marit Kramski, Thakshila H. Amarasena, Sheilajen Alcantara, Shayarana Gooneratne, Gilda Tachedjian 2,3, Robert De Rose, Stephen J. Kent, Wendy R. Winnall The University of Melbourne, Department of Microbiology and Immunology, Parkville, Australia, 2 Burnet Institute, Melbourne, Australia, 3 Monash University, Department of Microbiology, Clayton, Australia Background: Cameroon has one of the most genetically diverse HIV epidemics in the world. The country is home to virtually every known HIV subtypes, plus a vast array of CRFs and URFs. In the analysis we performed in a previous study (Tongo et al., 203), many of the Cameroonian CRFs and URFs showed up as having less evidence of recombination than many of the pure subtype sequences. This is exactly as would be expected if these Cameroonian sequences were extant examples of early diverging lineages or rare pre-epidemic HIV- group M lineages. Methods: Nine potential candidate ancient viruses were selected from our previous study and their full length sequences were generated. A maximum likelihood phylogenetic tree was constructed with 00 boostrap replicates with these sequences and a representative selection of the sequences from the rest of the world and all other published sequences from west central African countries. A blinded fully exploratory screen for recombination will be performed using RDP4. In addition, we wish to better understand the pathogenicity of these viruses by studying the function of nef and other accessory proteins. Results: Preliminary analyses revealed that the full genome sequence of one sample (BS3) was an outlier of all the CRF02_AG sequences sampled to date. Two full length sequences, BS and BS55, were both too divergent to be placed within any existing subtype or CRF grouping and therefore remained unclassified. They may in fact belong to a novel CRF or subtype, since they clustered with three other previously characterised Cameroonian sequences. The full length viral genome of BS72 was also too divergent to be placed within any existing subtype or CRF grouping. Analyses are underway for more phylogenetic and pathogenicity characterization. Conclusions: These preliminary results suggest that there is unquestionably a far more diverse pool of HIV sequences circulating amongst humans than presently accounted for in the current HIV- group M classification. Background: HIV drug resistance is a major problem, leading to poor outcomes for patients and rising treatment costs. Nucleoside/nucleotide (NRTI) resistance mutations often occur in reverse transcriptase (RT), resulting in either decreased incorporation or increased excision of NRTIs. RT mutations such as K65R, which confers tenofovir-resistance, increase the fidelity of RT while reducing viral fitness. To better understand the spread of drug-resistant HIV, we have used the pigtail macaque model to study the infectivity and fitness of the drug-resistant SIV mutants in vivo. Methods: We introduced the common K65R and the rare Q5N mutations into conserved regions of an SIV mac239 plasmid. CEM-NKr. CCR5 cells were infected with virus produced by transfected HEK-293T cells. Pigtail macaques were exposed to wildtype (wt) or mutant SIV by i.v. lethal dose of virus. Infection was monitored using qpcr, sequencing, western blotting for SIV proteins and ELISA for seroconversion. Results: SIV-K65R, but not SIV-Q5N, was able to infect CEM-NK5. CCR5 cells in vitro, although its replicative capacity was significantly lower than wt SIV, demonstrating decreased viral fitness in vitro. Pigtail macaques were exposed to wt SIV (n=3), SIV-K65R (n=2) or SIV-Q5N (n=2). Animals exposed to wt SIV became infected, with plasma SIV RNA peaking at day 4 and SIV-specific seroconversion detected at week 7. Although the SIV-K65R mutant established infection in macaques, it rapidly reverted to wt by day 4, before CTL and antibody responses developed. The SIV-Q5N mutant failed to infect macaques. Conclusions: Our results indicate that () although the drug-resistant SIV-K65R mutant has poorer replicative capacity in cell culture, it could establish infection in vivo before quickly reverting to a drug-sensitive infection in our animal model, (2) the high fidelity drug resistant SIV- Q5N mutant could not establish an SIV infection of macaques, suggesting poor transmissibility between humans. 272 HIV Research for Prevention 204 HIV R4P

273 Posters 25: Transmission and Viral Diversity P25. No Selection for Env with Shorter Variable Loops in Acute HIV- Infection Wednesday, 29 October Morgane Rolland, Sodsai Tovanabutra, Eric Sanders-Buell, Meera Bose, Anne Marie O Sullivan, Shana Howell, Kultida Poltavee, Jenica Lee, Grace Ibitamuno, Sevan Muhammad, Bahar Ahani, Steven Lepore, Elizabeth Harbolick, Celina Oropeza, Joey Patterson, Adam Bates, Michelle Lazzaro, Gustavo Kijak, Kalpana Dommaraju, Christopher Herr, Leigh Anne Eller, Sorachai Nitayaphan 2, Kathleen Rono 3, Lucas Maganga 4, Arthur Sekiziyivu 5, Nelson Michael 6, Jerome Kim 6, Merlin Robb MHRP;HJF, Silver Spring, MD, United States, 2 AFRIMS, Bangkok, Thailand, 3 WRP, Kericho, Kenya, 4 MMRP, Mbeya, Tanzania, United Republic of, 5 MUWRP, Kampala, Uganda, 6 MHRP, Silver Spring, MD, United States Background: Previous reports showed that HIV- viruses from early infection had shorter HIV- Env variable loops and could be more sensitive to neutralization. However, findings on HIV- subtype C and A were not confirmed with subtype B and there were uncertainties as to whether loop lengths evolved in the first weeks of infection. Here we characterized Env variable loops (V-V5) in subjects newly infected (diagnosis at a median of 4 days after the last negative visit) with different HIV- subtypes. Methods: We sequenced,204 HIV- strains from 49 subjects enrolled in Kenya, Tanzania, Uganda and Thailand at a median of 4, 33 and 7 days after diagnosis. These sequences were compared to 624 independent sequences from chronic infection. Results: Reflecting the distribution of subtypes in Thailand and East Africa, our cohort included individuals infected with CRF0_AE (n= 5), subtype A (n= 0), C (n= 4), and different A/C/D recombinant strains (n= 7). V loops varied between 8 and 34 amino acid (AA) (IQR = 6-23), while V2 loops varied between 36 and 5 AA (IQR = 40-44). Variable loop lengths did not differ between HIV- subtypes (p > 0.230). Next, we compared loop lengths from our cohort to values from chronically infected subjects: for 280 sequences isolated past 2000, V loops varied between 5 and 42 AA (IQR = 8-24), and V2 loops between 36 and 74 AA (IQR = 40-45). There was no evidence that variable loops from acutely-infected individuals were shorter than those from chronically-infected individuals (p > 0.88). Finally, we found that Env variable loop lengths did not increase over the first six months of follow up in our cohort (p > 0.352). Conclusions: Env sequences sampled in the first week of HIV- infection showed a wide range of variable loop lengths, making them undistinguishable from sequences from chronic infection. Our findings indicate that viruses with shorter Env variable loops are not selected for in the establishment of HIV- infection

274 Posters Posters 26: Vaccine Clinical Trials P26.0 P26.02 Quality Control Tool for Specimen Shipment in HIV Prevention Trials Avika Haridutt, Nathlee Abbai, Rashika Maharaj Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: In 2006 The Medical Research Council (MRC) HIV Prevention Research Unit (HPRU) set up a Repository for archiving specimens for various clinical trials which included Microbicide, PrEP and future Vaccine trials. The Repository currently houses samples. To ensure effective and efficient specimen management, the laboratory instituted the use of an electronic Laboratory Data Management System (LDMS). However, various challenges have been encountered during the specimen storage process. The following were identified amongst others as main errors: a) deviations of freezer temperatures as a result of manual monitoring of temperatures, which compromised specimen stability and b) samples being omitted in error for shipments. Methods: In 202, the introduction of a continuous temperature monitoring (CTM) system was necessitated which notified all relevant staff of temperature deviations at any given time to ensure specimen quality and integrity was not compromised within and external to business hours. In order to eliminate errors during shipment, a LDMS quality control (QC) tool was implemented by checking the shipping list against the shipping manifest as well as the physical sample to ensure that all specimens requested were shipped. Results: The introduction of the CTM led to a 00% notification of temperature deviations over the last 2 years. No samples were compromised for any study endpoints. The LDMS QC tool has shown on average a 00% reduction in the QC error rate over the last 2 years Conclusions: A high quality of specimens was maintained since the implementation of the CTM system from May 202 to date. The quality control process introduced for shipments eliminated duplication and reduced shipping costs. Lessons learnt from the challenges above has deemed the repository to an award winning status and has since been the benchmark for other institutions. Phase Trial of the Safety and Immunogenicity of PENNVAX -G DNA Prime Administered by Biojector 2000 or CELLECTRA EP Device with MVA-CMDR Boost Julie Ake, Silvia Ratto-Kim, Alexandra Schuetz 2, Lindsay Wieczorek, Viseth Nguay, Hannah Kibuuka 3, Fredrick Sawe 4, Leonard Maboko 5, Victoria Polonis, David Weiner 6, Mark DeSouza 2, Arthur Sekiziyivu 3, Josphat Kosgei 4, Marco Missanga 5, Arne Kroidl 5,7, Patricia Earl 8, Bernard Moss 8, Elizabeth Adams 9, Ana Martinez 9, Farrukh Rizvi 0, Amir Khan, Niranjan Sardesai, Nelson Michael, Mary Marovich 9, Merlin Robb, RV 262 Study Group US Military HIV Research Program/WRAIR, Silver Spring, MD, United States, 2 Armed Forces Research Institute of Medical Sciences, Department of Retrovirology, Bangkok, Thailand, 3 Makerere University - Walter Reed Project, Kampala, Uganda, 4 KEMRI/Walter Reed Project, Kericho, Kenya, 5 Mbeya Medical Research Centre, Mbeya, Tanzania, United Republic of, 6 University of Pennsylvania, Philadelphia, PA, United States, 7 University of Munich, Department for Infectious Diseases & Tropical Medicine, Munich, Germany, 8 Laboratory of Viral Diseases, NIAID, NIH, Bethesda, MD, United States, 9 Division of AIDS, NIAID, NIH, Bethesda, MD, United States, 0 Military Infectious Diseases Research Program, Ft. Detrick, MD, United States, Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States Background: Safety and immunogenicity were examined in a double blind, placebo controlled trial of PENNVAX -G (PVG) DNA, administered by Biojector 2000 (BJ) or CELLECTRA electroporation device (EP), boosted by Modified Vaccinia Ankara-Chang Mai Double Recombinant (MVA-CMDR) in healthy HIV uninfected adults. Methods: Subjects were randomized to receive 4 mg PVG DNA (4 plasmids containing consensus A, C, and D env sequences and a multiclade consensus gag) delivered IM by BJ or EP at baseline and month followed by IM injection of 0 8 pfu MVA-CMDR (expressing CRF0_AE CM235 env and subtype A CM240 gag/pol) at 3 and 6 months. The open label Part A (N=, US) was followed by the doubleblinded Part B (N=77, East Africa). Part B was randomized to BJ or EP and to placebo or PVG DNA/ MVA-CMDR (:4). For Part A and 6 Part B subjects, INF-γ ELISPOT responses were measured to vaccine env, gag, and pol inserts at baseline, 2 weeks post 2nd DNA injection and 2 weeks post 2nd MVA-CMDR injection. For Part A subjects, ELISA binding antibody (bab) endpoint titers and neutralizing antibody (TZMbl and PBMC) responses were measured at baseline, and 2 weeks post 2nd MVA, and at month 2. Part B remains blinded. Results: Study injections were safe; all 2 related AEs were mild or moderate. IFN-γ ELISPOT results were similar in BJ and EP groups. No responses were detected post DNA. Two weeks post 2 nd MVA the majority of responses were to env (7/4 BJ and 5/3 EP) followed by gag (2/4 BJ and 2/3 EP) with none to pol. 6/6 BJ and 4/5 EP recipients developed p24 bab responses peaking two weeks post 2nd MVA, persisting at month 2 in 5/6 BJ and 3/5 EP recipients. All subjects developed bab to gp20 (CM243), however responses waned in the majority by month 2. Neutralizing antibodies were detected in the PBMC assay using a CRF0_AE CM235 infectious molecular clone (6/6 BJ and 3/5 EP). Conclusions: Preliminary data demonstrate this PVG/MVA-CMDR regimen to be safe with no significant difference between DNA delivery methods. 274 HIV Research for Prevention 204 HIV R4P

275 Posters 26: Vaccine Clinical Trials P26.03 Wednesday, 29 October P26.04 Overweight BMI and Alcohol Use Are Associated with Immune Responses in Phase I HIV Vaccine Trials Detection of Vaccine Induced Mucosal Antibodies in Phase I HIV Preventative Vaccine Trials Roger Bayingana, Gaudensia Mutua 2, Juliet Mpendo 3, William Kilembe 4, Gloria Omosa-Manyonyi 2, Eugene Ruzagirwa 5, Liesl Page-Shipp 6, Glenda Gray 7, Linda-Gail Bekker 8, Lindsey Baden 9, Peter Hayes 0, Len Dally, Claudia Schmidt 2, Dagna Laufer 2, Frances Priddy 2, Pat Fast 2, Jill Gilmour 0, Josephine Cox 0 Projet San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 2 KAVI-Institute of Clinical Research, University of Nairobi, Nairobi, Kenya, 3 Uganda Virus Research Institute-IAVI, Entebbe, Uganda, 4 Zambia Emory HIV Research Program, Lusaka, Zambia, 5 Medical Research Council/Uganda Virus Research Institute, Uganda Research Unit on AIDS, Entebbe, Uganda, 6 Aurum Institute, Klerksdorp, South Africa, 7 Perinatal HIV Research Unit, Soweto, South Africa, 8 Desmond Tutu HIV Foundation, Cape Town, South Africa, 9 Brigham and Women s Hospital, Boston, MA, United States, 0 IAVI Human Immunology Lab, London, United Kingdom, EMMES Corporation, Rockville, MD, United States, 2 IAVI, New York, NY, United States Background: Biological or behavioral factors may impact vaccine immunogenicity and efficacy. We investigated the relationship between trial volunteer baseline demographic and behavioral factors and vaccinespecific immune responses in 3 phase HIV vaccine clinical trials. Methods: Volunteers at low-risk for HIV infection received 2-4 injections of candidate HIV vaccines over 3 to 6 months in 3 phase HIV vaccine clinical trials in Kenya, Uganda, Rwanda, Zambia, South Africa and the US from Vaccine recipients in trial arms with robust IFN-g ELISPOT responses at 4 weeks after the last vaccination were included in the analysis. Multivariate logistic regression model with backward selection was used to assess the relationship between parameters at baseline (gender, age, body mass index (BMI), alcohol use, drug use) and the odds of a positive IFN-g ELISPOT, defined as >38 spot forming cells/ml and >4 fold mean background. Results: 284 volunteers receiving at least one vaccine dose were included in the analysis. Of these, 44% were female, 54% age 26, mean BMI 23. (range 4-48). 3% reported weekly or daily alcohol use. Self-reported drug use was rare. Overweight BMI ( 30) correlated with 78% decreased odds of a positive ELISPOT response, p< When analyzed as a continuous variable, one unit increase in BMI correlated to a 8% decrease in the odds of a positive ELISPOT response, p< Daily or weekly alcohol use, compared to no alcohol use correlated with 3-fold increase in the odds of positive ELISPOT responses (p< 0.05). Age, gender, and any drug use were not associated with ELISPOT responses. Conclusions: Overweight BMI appears to be associated with decreased immune response to certain HIV vaccine candidates when measured by IFN-g ELISPOT in healthy, HIV low-risk populations. This effect may impact interpretation of phase trials which typically have small group sizes. The positive association with alcohol use deserves further analysis. Phil Bergin, Robert Langat 2, Bashir Farah 2, Simon Ogola 2, Gloria Omosa-Manyonyi 2, Gaudensia Mutua 2, Gina Outtara 2, Harriet Park 3, Jennifer Lehrman 3, Irene Mwangi 2, Helen Coutinho, Paramesh Chetty 4, Martin McMorrow, Josephine Cox, Pat Fast 3, Dagna Laufer 3, Jill Gilmour, Omu Anzala 2 IAVI Human Immunology Laboratory, London, United Kingdom, 2 Kenya AIDS Vaccine Initiative Institute of Clinical Research, Nairobi, Kenya, 3 International AIDS Vaccine Initiative, New York, NY, United States, 4 International AIDS Vaccine Initiative, Johannesburg, South Africa Background: Mucosal HIV-specific antibodies may help to prevent acquisition of HIV-. An efficacious vaccine may require a mucosal immune response. We show that vaccine-induced antibodies can be found in mucosal fluids, albeit at low frequency. Methods: The Kenya AIDS Vaccine Initiative (KAVI) took part in 3 Ph HIV vaccine trials (B002, B003 and B004) at 2 sites in Nairobi. Participants were consented for a mucosal sub-study and asked to provide saliva (SA), oral fluid (OF), semen (SM), cervico-vaginal (CV) and rectal (RC) specimens. Specimens were collected at baseline and 2 post-vaccination time points and IgG and IgA against either HIV- Gag (p24; B002 and B004) or Env (gp40; B003 and B004) were measured by ELISA, samples giving a positive reading (by absorbance) were then titrated. IgA and IgG antibodies in serum or plasma (SPL) were also measured at the same time points. Concordance was confirmed between assays at KAVI and the IAVI Human Immunology Laboratory, London. Results: Of 05 Kenyan participants, 89 consented to the sub-study and provided at least one mucosal sample type. Anti-HIV IgG and/ or IgA antibodies tested, to date, were detected in 32/93 (7%) oral fluid/parotid saliva, 3/42(3%) SM, 2/82 (5%) CV, 8/27(30%) RC and 68/84 (8%) of SPL samples. In SA, OF, CV and SM samples most antibody responses were IgG, while there were more volunteers with IgA in the RC samples. Some RC samples had low levels of IgA antibody reactivity prior to vaccination. Antibody titers at the mucosal surface ranged from :20-:00 while corresponding serum levels were up to a titer of :2500. Conclusions: It is possible to detect vaccine-induced antibodies against HIV at the mucosal surface. When antibodies are found in the mucosa, they are at much lower titers than those seen in the periphery

276 Posters Posters 26: Vaccine Clinical Trials P26.05 P26.06 Phase I Clinical Trial HIV-CORE002 of a Universal T-cell Vaccine: Mapping of CD8+ T Cell Epitopes Nicola J. Borthwick, Tina Ahmed, Lucy Dorrell 2, Andy Van Hateren 3, Tim Elliot 3, Tomas Hanke Oxford University, Jenner Institute, Oxford, United Kingdom, 2 Oxford University, Nuffield Department of Medicine, Oxford, United Kingdom, 3 Southampton University, Southampton, United Kingdom Background: HIV-CORE002 was a phase I clinical trial of HIVconsv, an immunogen composed of conserved regions of the HIV- proteome and vectored by plasmid (DNA; D), modified vaccinia virus Ankara (MVA: M) and non-replicating chimpanzee adenovirus (ChAdV-63; C) in heterologous prime-boost vaccine regimens CM, DDDCM and DDDMC. The CM & DDDCM regimens were highly immunogenic and crucially CD8+ T lymphocytes suppressed HIV- replication in an in vitro virus inhibition assay. In this study we have mapped the CD8+ T cell epitopes induced by vaccination and have investigated their anti-viral function. Methods: A panel of 99 5-mer peptides overlapping amino acids across the length of HIVconsv were used to identify the top 5 responses per person. Panels of truncated peptides were used to define minimal epitopes. Results: In the CM arm of the trial, we initially divided the peptides into CD4 and/or CD8 cell subsets based on IFN-γ production. 42.8% of the 5-mer peptides induced a single CD8+ response and 39.2% of peptides induced responses in both subsets. Using the participants HLA type combined with previously identified and/or predicted epitopes, 34 putative CD8+ epitopes, mainly within Pol, were identified. All of the CD8+ T cells released IFN-γ, TNF-α and CD07a following peptide stimulation indicating they are oligofunctional effectors. Restriction and anti-viral activity of selected peptides are being investigated. Of particular interest was an immunodominant epitope within peptide 93 recognised by everyone sharing an HLA A02 supertype.. The epitope mapped as YQYMDDLYV and is located within the active site of reverse transcriptase. Using a competition binding assay the peptide binds with high affinity to HLA-A*02:0. Cell lines generated using peptide 93 were cytotoxic against autologous peptide pulsed targets and the anti-hiv- activity will be presented. Conclusions: Mapping of epitopes and functionalities of corresponding vaccine-elicited T cells will inform further vaccine development. A Phase I Clinical Trial with a Novel gp4 HIV Vaccine (EN4-FPA2) in Healthy Female Volunteers: A Mucosal Prime and Intramuscular Boost Regimen Alethea V. Cope, Christiane Moog 2, Robin J. Shattock, Mira M. Chawda, Justyna Czyzewska-Khan, Pizzoferro Kat 3, Suzanne Venables, Celine Yan, Marie Williams 3, Karen Cobb 3, Mahavir Singh 4, Wulf Oehlmann 4, Ayssar Elamin 4, Dietmar Katinger 5, Andreas Wagner 5, Paulatsya Joshi 3, David J.M. Lewis 3 Imperial College, Medicine, London, United Kingdom, 2 Inserm, Strasbourg, France, 3 University of Surrey, Clinical Research Centre, Guildford, United Kingdom, 4 Lionex GmbH, Braunschweig, Germany, 5 Polymun Scientific GmbH, Klosterneuburg, Austria Background: We conducted a randomised single-centre, observer blind trial with a fourth generation gp4 vaccine (EN4-FPA2) designed to expose 2F5 and 4E0 epitopes formulated as liposomes. Methods: HIV negative female volunteers aged 8-55 years were randomized into groups to receive 3 nasal primes at weeks 0, 4 and 8: () Low dose 20µg (2) Mid-dose 00mg (3) Full dose 200mg. Groups -3 & group 4 (nasal placebo) received two intramuscular boosts at weeks 2 and 20, group 5 was double placebo. We measured immunogenicity; B cell responses in serum, mucosal samples by ELISA, memory B-cell Elispot and neutralization. T cell responses were analysed by Elispot and ICS. Gene array analysis was also conducted to investigate the role of intranasal priming on innate immune factors and signalling. Results: 48 participants were enrolled in total the vaccine was safe and well tolerated via both routes with no related serious adverse events. Primary immunogenicity was achieved in all participants regardless of priming dose. At week 24 a median specific serum IgG of 7.94µg/ml ( ) vs 2.50µg/ml ( ) was detected (group 3 & 4, respectively). Serum IgA was detected in 48% of participants with a median.94µg/ml ( ). Mucosal IgG was detected in 66% of women, median 0.26µg/ml ( ) which exhibited low-level inhibitory activity. No significant differences in memory B cell responses and no nasal priming effect for T cells by ICS were observed. Vaccineinduced memory B and T cell responses were detected in the periphery without evidence of mucosal nasal priming. However, mucosal priming was evident by gene array analysis where innate factors and immune pathways were up-regulated. Conclusions: The gp4 vaccine was safe, well tolerated and immunogenic with potent serum responses detected and mucosal responses that exhibited low-level inhibitory activity. Furthermore vaccine induced up-regulation of innate factors by nasal priming could be important in future effective vaccine design approaches. 276 HIV Research for Prevention 204 HIV R4P

277 Posters 26: Vaccine Clinical Trials P26.07 Assessment of Viral Inhibition Activity in Low Seroprevalent Adenovirus-35 Vectored HIV Vaccines +/- Adjuvanted Protein or Electroporated DNA Wednesday, 29 October P26.08 Immune Responses after Two Immunizations with rgp40/gla Following Priming with HIV-DNA and HIV-MVA in Healthy Tanzanian Volunteers Peter Hayes, Natalia Fernandez, Gloria Omosa-Manyonyi 2, Juliet Mpendo 3, Etienne Karita 4, Eugene Ruzagira 5, William Kilembe 5, Gaudensia Mutua 6, Omu Anzala 6, François Roman 7, Patricia Bourguignon 7, Burc Barin 8, John Eldridge 9, Michael Egan 9, Drew Hannaman 0, Claudia Schmidt, Patricia Fast, Frances Priddy, Josephine Cox, Jill Gilmour International AIDS Vaccine Initiative (IAVI)-Human Immunology Lab, Imperial College, London, United Kingdom, 2 Kenya AIDS Vaccine Initiative Institute of Clinical Research, Nairobi, Kenya, 3 Uganda Virus Research Institute-IAVI, Entebbe, Uganda, 4 Project San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 5 Zambia Emory HIV Research Program, Lusaka, Zambia, 6 Kenya AIDS Vaccine Initiative- Kangemi, University of Nairobi, Nairobi, Kenya, 7 GlaxoSmithKline Vaccines, Rixensart, Belgium, 8 EMMES Corporation, Rockville, MD, United States, 9 Profectus Biosciences, Tarrytown, NY, United States, 0 ICHOR Medical Systems, San Diego, CA, United States, International AIDS Vaccine Initiative, New York, NY, United States Background: CD8 T-cells suppress HIV replication and this activity controls acute viremia and slows progression to AIDS. We have validated a functional ex vivo Viral Inhibition Assay (VIA) to measure virus suppression in HIV vaccine recipients. Methods: VIA, using a panel of 8 subtype A to D viruses, was compared in 45 vaccine recipients from two prime-boost trials: B002 trial assessed adjuvanted Gag-Pol-Nef Fusion Protein (F4/AS0) co- or sequentially administered with replication-defective Ad35 vector encoding clade A genes gag, RT, integrase, nef (Ad35-GRIN). B004 trial assessed plasmid DNA (HIVMAG) encoding clade B gag-pol, env, nef-tat-vif, administered by intramuscular electroporation alone or with pdna IL-2 and Ad35- GRIN and Ad35-Env. VIA assessed by the log reduction in HIV p24 production was measured at baseline, and 4 weeks after prime and boost immunizations. Results: VIA activity was detected in 3/9 vaccinees (68%) after priming with 3 HIVMAG +/- IL-2, and after 2 F4/AS0 administrations in 2/8 (25%) vaccinees. After Ad35-GRIN +/- Env boost, VIA activity was detected in 8/9 (95%) and 6/8 (75%) volunteers respectively. When F4/AS0 was co-administered with Ad35-GRIN, VIA was detected in 5/8 vaccinees (63%) after the st and 7/9 (78%) after the 2 nd and 3 rd administrations. Clade B (IIIB) and A (U455) viruses were inhibited most frequently. In both trials, at 4 weeks after Ad-HIV vaccine, VIA activity was similar in magnitude and in breadth, assessed by the number of viruses inhibited. The mean log inhibition for responders was 2.7 (range.5-4.8) and an average of 3 viruses were inhibited after Ad35-GRIN +/-Ad35-Env. VIA from these 2 trials will be compared with VIA induced by other vaccines and regimens. Conclusions: Modest frequency and magnitude VIA response to electroporated HIVMAG or F4/AS0 was significantly boosted by combination with Ad35-GRIN+/- Ad35-Env. Thus Ad35 provides a suitable platform for induction of functional CD8-T cells with breadth and potency. Agricola Joachim,2, Asli Bauer 3,4, Sarah Joseph 5, Christof Geldmacher 4,6, Charlotta Nilsson 2,7, Patricia Munseri 8, Marko Missanga 3, Philip Mann 3,4, Said Aboud, L Sudi 3, P. Mviombo 3, A. Haule 3, Britta Wahren 2, Guido Ferari 9, Vicky Polonis 0, Merlin Robb, Roger Tatoud 2, Leonard Maboko 3, Eligius F. Lyamuya, Michael Hoelscher 4,6, Muhammad Bakari 8, Gunnel Biberfeldt 2,7, Eric Sandstrom 3, Arne Kroidl 4,6, Sheena Mc Cormack 5 Muhimbili University of Health and Allied Sciences, Microbiology and Immunology, Dar es Salaam, Tanzania, United Republic of, 2 Karolinska Institutet, Microbiology, Tumor and Cell Biology, Stockholm, Sweden, 3 NIMR Mbeya Medical Research Center, Dar es Salaam, Tanzania, United Republic of, 4 Klinikum of the University of Munich, Infectious Disease and Tropical Medicine, Munich, Germany, 5 MRC Clinical Trials Unit at University College London, London, United Kingdom, 6 German Center for Infection Research, Munich, Germany, 7 Public Health Agency of Sweden, Solna, Sweden, 8 Muhimbili University of Health and Allied Sciences, Internal Medicine, Dar es Salaam, Tanzania, United Republic of, 9 Duke University Medical Center, Surgery, Durham, NC, United States, 0 Walter Reed Army Institute of Research, Silver Spring, MD, United States, Military HIV Research Program, Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 2 Imperial College London, London, United Kingdom, 3 Venhälsan, Karolinska Institutet at Södersjukhuset, Stockholm, Sweden Background: We evaluated the effects of two adjuvanted clade C Env protein immunizations after multi-clade HIV-DNA and HIV-MVA priming in healthy Tanzanian volunteers. The DNA included a plasmid encoding Clade C Env. Methods: Thirty-five volunteers primed three times with HIV-DNA encoding HIV- subtypes A, B, and C and boosted twice with MVA CMDR expressing CRF0_AE were further boosted with two doses of trimeric CN54rgp40 subtype C adjuvanted with GLA-AF (IDRI; rgp40/ GLA). Five placebo recipients received the same immunizations with rgp40/gla. Antibody (Ab)- and cell-mediated immune responses were assessed. Results: Boosting DNA/MVA vaccine recipients (n=35) twice with rgp40/gla increased median anti-cn54gp40 IgG titers considerably from 900 to (p< 0.000). After rgp40/gla boosts DNA/MVA vaccinees had higher median rgp40 IgG titers (24300) compared to placebo recipients (2700, p=0.036) and those with elevated titers of 900 after the DNA/MVA prime had higher titers after the first (p=0.0009) and second rgp40/gla boost (p=0.0449). No significant increase in neutralizing antibody activity was seen in an infectious molecular clone (IMC)/PBMC assay against GS05 subtype C or CM235 CRF0_AE after the second rgp40/gla. Only one vaccinee developed antibody-dependent cellular cytotoxicity-ab to HIV 086 subtype C IMC after two rgp40/gla vaccinations. The IFN-g ELISpot response rate to Env increased from 29.4% (0/34) at the time of the first rgp40/gla to 68.6% (24/35) after the second. Conclusions: DNA/MVA priming had a significant effect on anti- CN54gp40 IgG responses after boosting with rgp40/gla. Immunization with rgp40/gla enhanced binding Ab responses and cell-mediated immune responses in HIV-DNA/MVA-primed Tanzanian vaccines, but no significant increase in functional humoral responses could be detected

278 Posters Posters 26: Vaccine Clinical Trials P26.09 Transgender Participants in the HIV Vaccine Trial Network s HVTN 505 Trial: A Descriptive and Comparative Analysis P26.0 Update of the Long-term Follow-up of Healthy Volunteers from Preventive HIV- Vaccine Trials: ANRS COV-COHVAC Cohort Shelly T. Karuna,2, Doug Grove, Gail B. Broder,3, Chuka Anude 4, Scott M. Hammer 5, Magda E. Sobieszczyk 5, Michele P. Andrasik,2, HVTN 505 Protocol Team of the NIAID-funded HIV Vaccine Trials Network Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 HIV Vaccine Trials Network, Clinical Development Unit, Seattle, WA, United States, 3 HIV Vaccine Trials Network, Community Engagement Unit, Seattle, WA, United States, 4 DAIDS/NIAID/NIH, Vaccine Clinical Research Branch, Washington, DC, United States, 5 Columbia University College of Physicians and Surgeons, Division of Infectious Diseases, Department of Medicine, New York, NY, United States Background: HIV prevalence in the US transgender (TG) population (2.64%) is five times the national average (0.45%), yet TG individuals constitute a small proportion of HIV vaccine trial participants (ppts). HVTN 505, a Phase 2b trial to assess the safety and efficacy of a DNA/ rad5 HIV vaccine regimen, was the HVTN s first trial to specify the eligibility of TG women born male, distinct from MSM. Methods: Data was analyzed for the HVTN 505 modified-intent-to-treat (MITT) cohort (n=2496), which included ppts who were HIV-uninfected at enrollment. To identify factors that may impact future engagement, enrollment, and retention, ppts reporting gender identity differing from birth sex (TG) were compared to ppts reporting concordant gender identity and birth sex (cisgender, CG). Results: Forty-four (.8%) TG ppts were compared to 2452 (98.2%) CG ppts in the MITT cohort. Of TG ppts, 4% identified Hispanic, 43% White, 4% African-American, % other, and 2% each as Asian or multi-racial. TG ppts were more likely to be non-white (57% TG vs. 24% CG; p<.00) and younger (mean 29 vs. 32 years old; p=0.04). TG and CG ppts did not differ in average number of pre-existing conditions (PECs) or adverse events (AEs), including total serious AEs. There were more deaths among TG ppts (n=2 or 5% vs. n=7 or 0.29%; p=.0) and more TG ppts missed at least one study visit (34% vs. 2%; p=.0). There were no differences in numbers of social harms or benefits, or in numbers of interim HIV tests. Differences in HIV incidence were not statistically significant (3 TG ppts or 4.3/00 person-yrs vs. 84 CG ppts or 2./00 p-y; p=0.37). Conclusions: HVTN 505 TG ppts were more likely than CG to be characterized by demographic factors associated with HIV incidence, including non-white race and young age. Though overall HIV incidence was higher among TG ppts, the difference was not significant. Missed visits were more frequent among TG ppts; there was no difference in PECs and AEs. Potential implications for future trials will be presented. Corinne Desaint,2,3, Christine Durier 4, Jean-Daniel Lelièvre 5,6,7, Benjamin Silbermann 8, Gilles Pialoux 9, Lise Cuzin 0, Isabelle Poizot-Martin,2,3, Pascale Morineau 4, Amel Bouakane 5, Bruno Spire,6, Yves Lévy 5,6,7, Jean-Pierre Aboulker 4, Odile Launay,2,3, ANRS COHVAC Study Group, Paris, France and Vaccine Research Institute (VRI), Créteil, France Université Paris-Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France, 2 Inserm, CIC 47, Paris, France, 3 AP-HP, Hôpital Cochin, CIC de Vaccinologie Cochin Pasteur, Paris, France, 4 Inserm, SC0- US09, Villejuif, France, 5 Inserm, U955, Créteil, France, 6 Université Paris-Est Créteil Val de Marne (UPEC), Créteil, France, 7 AP-HP, Hôpital Henri Mondor, Service d Immunologie Clinique, Créteil, France, 8 AP-HP, Hôpital Cochin, Service de Médecine Interne, Paris, France, 9 AP-HP, Hôpital Tenon, Service des Maladies Infectieuses et Tropicales, Paris, France, 0 CHU Toulouse, Service des Maladies Infectieuses, Toulouse, France, Université Aix-Marseille, Marseille, France, 2 APHM Hôpital Sainte Marguerite, Service d Immuno-hématologie clinique, Marseille, France, 3 Inserm, U92 (SESSTIM), Marseille, France, 4 CHU Nantes, Service des Maladies Infectieuses, Nantes, France, 5 ANRS, Service de Recherche Vaccinale, Paris, France, 6 Inserm / IRD, UMR 92, Marseille, France Background: Long-term epidemiological studies of safety of vaccine candidates are scarce. ANRS COV-COHVAC cohort is a prospective cohort study set up to describe the long-term safety of preventive HIVvaccines administered in 7 phase I and II clinical trials. Methods: All 496 ANRS volunteers (healthy, aged 2-55, at low risk for HIV, no financial incentive) were eligible to retrospective collection starting from the first injection of a candidate vaccine and to prospective 7-years annual follow-up (FU). Health events evoking neurological, ophtalmological and immunological disorders (any grade) and other grade 3/4 events are recorded and consequences of vaccine induced seropositivity (VISP) are also studied. A self-administered questionnaire is filled in at each visit. Results: From 2009 to 202, all but 8 participants were enrolled either for prospective (n=355, 73%) or for retrospective study only (n=33, 27%). At trial entry, median age was 43 years (23 to 56), 55% were men. As of March 204, median total FU after vaccination was 7.3 years (0.5 to 20.8) and 3 years (0 to 4.6) in the prospective cohort with 3 (9%) dropouts. At enrolment, 5 years in median after vaccination (2 to 8), 45 (3%) reported psycho-social difficulties to the investigator while 38 (%) self-reported trial-related problems, mostly with spouse and family. Unprotected intercourse with casual partner in the past year was reported by 7 participants (5%). Six (2%) reported unsafe sex with HIV-positive or unknown status partners. At last prospective FU (mostly 3-4 years), VISP was persistent in 7 participants out of 29 who received recombinant HIV- envelope protein, 8-20 years ago. There was incident HIV infection. Validation of clinical events is ongoing; no safety signals have been identified yet. Conclusions: In ANRS COHVAC cohort, low rate of trial-related issues and stable VISP status are confirmed. High FU rate will allow thorough evaluation of clinical safety of HIV candidate vaccines. 278 HIV Research for Prevention 204 HIV R4P

279 Posters 26: Vaccine Clinical Trials P26. Wednesday, 29 October P26.2 Evaluation of the Immunogenicity and Impact on the Latent HIV- Reservoir of an HIV Conserved Region Vaccine, MVA.HIVconsv, in HAART-treated Subjects Sara Morón-López, Gemma Hancock 2, Maria Carmen Puertas, Annie Rose 2, Maria Salgado, Emma-Jo Hayton 2, Catharine Morgan 3, Brian Angus 2, Hongbing Yang 2, Tomas Hanke 2, Lucy Dorrell 2,3, Javier Martinez-Picado,4 AIDS Research Institute IrsiCaixa, Badalona, Spain, 2 Oxford NIHR Biomedical Research Centre, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom, 3 Genitourinary Medicine, Oxford University Hospitals NHS Trust, Oxford, United Kingdom, 4 ICREA, Barcelona, Spain Major Negative Social Impacts Are Rare in Phase HIV Vaccine Trials in Africa Gaudensia Mutua, Lillian Mutengu 2, Juliet Mpendo 3, William Kilembe 4, Gloria Omosa-Manyonyi, Eugene Ruzagira 5, Etienne Karita 6, Liesl Page-Shipp 7, Glenda Gray 8, Linda-Gail Bekker 9, Burc Barin 0, Dagna Laufer, Claudia Schmidt, Frances Priddy KAVI-Institute of Clinical Research, University of Nairobi, Nairobi, Kenya, 2 IAVI, Nairobi, Kenya, 3 Uganda Virus Research Institute-IAVI, Entebbe, Uganda, 4 Zambia Emory HIV Research Program, Lusaka, Zambia, 5 MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 6 Projet San Francisco, Rwanda Zambia HIV Research Group, Kigali, Rwanda, 7 Aurum Institute, Klerksdorp, South Africa, 8 Perinatal HIV Research Unit, Soweto, South Africa, 9 Desmond Tutu HIV Foundation, Cape Town, South Africa, 0 EMMES Corporation, Rockville, MD, United States, IAVI, New York, NY, United States Background: New therapeutic vaccines are potential strategies to achieve a functional cure for HIV-. A novel conserved region immunogen, HIVconsv, elicited potent T cell responses in healthy subjects when delivered by a heterologous viral vector regimen comprising a simian adenovirus prime and MVA boost. We investigated whether MVA.HIVconsv alone could induce T cell responses of sufficient potency to impact the latent HIV- reservoir in HAART-treated subjects. Methods: 9 chronically infected HAART-suppressed subjects were randomised to receive 3 intramuscular injections of MVA.HIVconsv high dose (2.2x0 8 pfu) (n = 7), MVA.HIVconsv low dose (5.5x0 7 pfu) (n = 8), or placebo (n = 4) at 0, 4 and 2 weeks. Cell-associated HIV- DNA in circulating CD4 + T cells was estimated by droplet digital PCR at weeks 0, 6 and 38 and ex vivo CD8 + T cell inhibition of HIV- in autologous CD4 + T cells at baseline and week 8. Results: 90% subjects completed the vaccine regimen. Baseline median [IQR] cell-associated HIV- DNA copies / 0 6 CD4 + T cells were: high dose [396-3,92]; low dose -,074 [78-2,50]; placebo -,652 [986-2,643] (p=0.505). Total HIV- DNA did not change significantly from baseline at either week 6 or at week 38 in any group (high dose - 72 [599-2,799] w6 and 577 [387-3,25] w38; low dose -,260 [424-2,498] w6 and,272 [793-,95] w38; placebos - 2,058 [72-4,060] w6 and,35 [594-,976] w38). CD8 + T cell viral inhibitory capacity (E:T ratio :) was low in all groups at baseline and a post-vaccination increase occurred only in the high dose group: medians pre- and postvaccination were: high dose - 7% vs. 54% (p = 0.06); low dose - 3% vs. 3%; placebos - 0% vs. 5%. Conclusions: Vaccination of chronic HAART-suppressed subjects with high dose MVA.HIVconsv induced a modest increase in CD8 + T cell antiviral activity but did not significantly change the size of the HIV- reservoir. More potent vaccine regimens, together with latency-reversing agents, may be needed. Background: Participation in HIV prevention trials may have important positive or negative social impacts for the volunteer. We assessed selfreported social impact in 3 phase HIV vaccine clinical trials in 5 African countries. Methods: Three phase HIV vaccine clinical trials in Kenya, Uganda, Rwanda, Zambia and South Africa enrolled volunteers at low-risk for HIV, who received 2-4 injections and were followed for up to 6 months with repeated HIV testing and counseling, plus mucosal sampling at the Kenyan sites. At the final study visit, self-reported data on potential social impact of trial participation was collected using a standardized questionnaire. Volunteers rated whether the impact was harmful or not, and graded harmful impacts as mild, moderate, major. Comparisons of categorical factors were conducted using the Fisher s exact test. A twosided p< 0.05 was considered statistically significant. Results: Social impact data was collected on 383 trial volunteers. 42% (62/383) reported one or more (256 total) social impacts. The majority (75/256, 68%) were reported as positive impacts, with the most common being affecting your feelings on the AIDS epidemic 58% (0/75), and affecting feelings about yourself 4% (24/75). The most common negative impacts were affecting health 33% (27/8) and affecting relationship with friends 7% (4/8). Negative social impacts were not more common in women (2%) than men (7%) (p=0.5), but varied by country (Uganda 28%, Kenya 22%, Zambia 8%, Rwanda 6%, South Africa 0%, p< 0.000). A total of 7 negative impacts were rated as major: low libido (4), falling sick frequently (), believed infected by HIV () and dental problems (); and 24 were rated as moderate. Conclusions: Trial volunteers reported largely positive social impacts and few major negative impacts after participation in phase HIV vaccine trials. These data will inform appropriate counseling messages in future phase and larger vaccine efficacy trials in Africa

280 Posters Posters 26: Vaccine Clinical Trials P26.3 P26.4 Vaccine Induced Seroreactivity in RV44 Vaccine Recipients in RV305, a Placebo Controlled Assessment of Late Boosts with ALVAC-HIV and AIDSVAX B/E Chirapa Eamsila, Siriwat Akapirat, Sorachai Nitayapan, Supachai Rerks-Ngarm 2, Punnee Pitisutthithum 3, Sandhya Vasan,4, Jean-Louis Excler 4, Nicos Karasavvas, Viseth Ngauy, Merlin L. Robb 4,5, Nelson L. Michael 5, Jerome H. Kim 5, Robert J. O Connell Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 2 Ministry of Public Health, Department of Disease Control, Nothaburi, Thailand, 3 Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand, 4 Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States, 5 US Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD, United States Background: HIV vaccine candidates induce immune responses intended to protect from infection but also cause vaccine induced seroreactivity (VISR) that can cause social harms and necessitates complicated algorithms to establish infection status. One year after last vaccination, VISR was detected by EIA in 0.% of RV44 vaccine recipients. Methods: Consenting RV44 vaccine recipients 6 to 8 years following last study vaccine were randomized to receive ALVAC-HIV plus AIDSVAX B/E, AIDSVAX B/E, or ALVAC-HIV in groups -3 respectively, or placebo, at weeks 0 and 24. HIV infection status was determined at screening and at weeks 0, 24, 48 and 72 using Genetic Systems (GS) HIV-/HIV-2 Plus O EIA. Repeatedly reactive samples underwent GS HIV- Western blot. End-of-study samples with EIA reactivity were tested with the Thai FDA-approved Enzygnost Anti-HIV /2 Plus EIA. Results: One hundred sixty volunteers completed all study visits, and all remained HIV-uninfected during the study. No EIA reactivity was detected at study entry or in any of the 27 placebo recipients. EIA reactivity occurred in at least one time point in 2/34(9%) vaccine recipients, 4/45(9%), 0/45(0%), and 8/43(9%), p<0.05, in groups, 2, and 3, including end-of-study reactivity in 7/33(5%) overall and 4/45(9%), 0/45(0%), and 3/43 (7%), p=ns, in groups, 2, and 3. All end-of-study samples reactive by GS EIA were non-reactive by Enzygnost EIA. All 27 Western blots produced an indeterminate result, with 00% band reactivity at p24, p5, and p55, and 0% reactivity at p8, p65, gp20, and gp60. Conclusions: Late boosts with ALVAC-HIV plus AIDSVAX B/E or ALVAC- HIV alone yielded end of study VISR detected by EIA in much larger proportion than seen in RV44, but did not induce positive Western blot results. The Enzygnost EIA did not cross react with reactive GS samples, suggesting its utility as a screening test for pox-protein vaccine recipients. Sex Differences in Immune Variables in the RV44 Trial Darpun D. Sachdev,2, Hasan Ahmed 3, Yunda Huang 3, Peter Gilbert 3, Supachai Rerks-Ngarm 4, Jaranit Kaewkungwal 5, Sorachai Nitayaphan 6, Nelson Michael 7, Punnee Pitisuttithum 8, Merlin Robb 9, Jerome H. Kim 0, Robert O Connell, Nicole Frahm 2, Cecilia Morgan 3, Susan P. Buchbinder 2,4 Center for AIDS Prevention Studies (CAPS), UCSF, San Francisco, CA, United States, 2 San Francisco Department of Public Health, Bridge HIV, San Francisco, CA, United States, 3 SCHARP - FHCRC, Seattle, WA, United States, 4 Thailand-MoPH U.S. CDC Collaboration, Department of Disease Control, Nonthaburi, Thailand, 5 Mahidol University, Bangkok, Thailand, 6 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 7 Walter Reed Army Institute of Research, U.S. Army Military HIV Research Program, Silver Spring, MD, United States, 8 Mahidol University, Vaccine Trials Center, Bangkok, Thailand, 9 Walter Reed Army Institute of Research, U.S. Army Military HIV Research Program, Bethesda, MD, United States, 0 Walter Reed Army Institute of Research, U.S. Military HIV Research Program, Silver Spring, MD, United States, Armed Forces Research Institute of Medical Sciences, U.S. Army Medical Component, Bangkok, Thailand, 2 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 3 Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 4 University of California at San Francisco, Department of Epidemiology and Biostatistics, San Francisco, CA, United States Background: In the RV44 vaccine trial there was a moderate, but not statistically significant difference in vaccine efficacy between males and females (25.8% v 38.6%, respectively.) Given known sex differences in immune responses to vaccines and HIV, we evaluated for sex differences in immune responses to RV44 vaccination to provide possible biological explanations. Methods: Participants were administered blinded placebo or 4 doses of prime (canarypox ALVAC-HIV: vcp52) and 2 doses of gp20 AIDSVAX B/E between weeks 0 and 24. We examined 97 immune variables for sex differences using week 26 data from uninfected vaccine recipients (60 females, 20 males) and 20 placebo recipients in the RV44 case-control cohort. We used two-sided Wilcoxon rank sum tests for the comparisons and the false discovery rates (q-value) to account for multiplicity. For variables with q< 0.2 we compared uninfected vaccinees to placebo recipients. Results: For 85 variables females and males had similar levels. No sex differences in humoral immune responses as measured by specific antibody levels were significant. Of 7 natural killer (NK) and dendritic cell (DC) phenotyping variables, males had higher mean percentages of CDc+ myeloid (M)DC of live peripheral blood mononuclear cells (p=2.8e-5, q-value=0.05), plasmacytoid (P)DC (p=2.9e-5, q-value=0.4), and CD6-CD56dim NK cells(p=.03e-3, q=0.07) compared to females. Innate immune measures were not significantly different in vaccinees vs. placebo recipients (p>0.25). The distributions of potential confounding factors such as age and risk behavior were similar across sexes. Conclusions: Male vaccinees were found to have increased percentages of MDCs, PDCs and CD6-CD56dim NK cells compared to females. Sex hormones and quantitative differences in innate immune cell types may affect the quality, quantity or longevity of immune responses to RV44 vaccination. 280 HIV Research for Prevention 204 HIV R4P

281 Posters 26: Vaccine Clinical Trials P26.5 Multivalent HIV DNA Vaccination to Simultaneous but Spatially Distinct Dites via an Electroporation Device Allows for Increased Dose Delivery Wednesday, 29 October P26.6 Phase I HIV Vaccine Trial to Assess Safety and Immunogenicity of DNA Priming and MVA Boosting in Healthy Mozambican Young Adults Kate E. Broderick, Jay B. McCoy, Janess M. Mendoza, Katherine Schultheis, Megan C. Wise 2, David B. Weiner 2, Laurent M. Humeau, Niranjan Y. Sardesai 3 Inovio Pharmaceuticals, Inc., San Diego, CA, United States, 2 University of Pennsylvania, Philadelphia, PA, United States, 3 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States Background: We have recently reported clinical trial data from two studies (HVTN-080 and HIV-00) showing the induction of robust T-cell responses to HIV immunogens (Env, gag, pol) when administered intramuscularly (IM) followed by electroporation (EP). The HVTN-080 study also demonstrated the benefit of co-delivering plasmid IL-2 together with the vaccine to drive strong T-cell responses. In parallel, we have reported on the use of intradermal (ID) EP to drive strong humoral responses in NHP and guinea pigs to HIV antigens. Methods: Here we report the development of a multi-headed intradermal electroporation device (msep) designed to deliver multiple DNA vaccine plasmids simultaneously to four spatially separated sites. In addition to increasing the vaccine dose-volume that can be delivered via ID application, the m4sep device allows for site specific tailoring of DNA vaccine delivery, where multiple different target vaccines may be delivered at once, and mitigating any potential issues with vaccine interference. Results: Reporter gene plasmids expressing GFP and RFP were used to demonstrate the impact of spatial separation on DNA delivery to increase the number of transfected cells and avoid interference. We demonstrate through immunogenicity read-outs that the multi-head device facilitates higher dose delivery to the skin resulting in over 2x improved antibody titers relative to single site delivery. Additionally, we demonstrate that a combination DNA vaccine, containing multiple HIV env constructs, can be delivered using the msep device and maintain humoral responses to each component vaccine. Conclusions: We conclude that the new multi-head DNA delivery device is an efficient, tolerable and non-invasive method to deliver multiple plasmid DNA constructs simultaneously The battery operated, low-cost, easy to use device platform for the delivery of multi-agent DNA vaccines has direct applications for prophylactic vaccination in mass vaccination settings. Edna Omar Viegas,2,3, Nelson Tembe,2,3, Charlotta Nilsson 2,4, Bindiya Meggi, Norma Mabota, Nádia Sitoe, Cremildo Maueia, Raquel Chissumba, Victória Cumbane, Eulália Macovela 3,5, Emília Gonçalves 3,5, Rick Stout 6, Gabriella Scarlatti 7, Britta Wahren 2, Soren Andersson 8, Mary Marovich 9, Merlin Robb 9, Nafissa Osman 3,5, Gunnel Biberfeld G 2,4, Ilesh Jani, Eric Sandström 2 Instituto Nacional de Saúde, Maputo, Mozambique, 2 Karolinska Institutet, Stockholm, Sweden, 3 Eduardo Mondlane University, Maputo, Mozambique, 4 Public Health Agency of Sweden, Stockholm, Sweden, 5 Hospital Central de Maputo, Maputo, Mozambique, 6 Bioject Inc., Tualatin, OR, United States, 7 IRCCS San Raffaele Scientific Institute, Milan, Italy, 8 Örebro University Hospital, Örebro, Sweden, 9 Walter Reed Army Institute of Research, Rockville, MD, United States Background: Safety and immunogenicity of intradermal HIV-DNA priming at doses of 600µg or 200µg followed by intramuscular HIV- MVA-CMDR boosts were evaluated in this study. Methods: Healthy volunteers (n=24), aged 8-26, recruited at a Youth Clinic in Maputo, Mozambique, were randomized into two groups: 0 volunteers received 600µg (0.ml) or 200µg (0.2ml), respectively, of DNA intradermally using a needle-free device, Zetajet (M0, M, and M3), followed by MVA boosts (M6 and M9), and four received placebo. Safety endpoints were assessed clinically and by standard chemistry and haematology tests. Primary immunogenicity endpoint was determined by IFN- γ ELISpot. Results: The ratio of screening and enrolment was 3:.The main reasons for screening failures were risk behaviour, abnormal CBC and positive syphilis tests. During the trial, 2 volunteers became pregnant and was infected with HIV. None of them had completed their vaccination schedule. HIV infection was the only Serious Adverse Event registered. Two grade 3 and 2 grade 4 laboratory adverse events were registered without probable relationship to vaccination. After the first MVA, 4/5 (93%) of vaccinees were IFN-γ ELISpot reactive:3 (87%) to Env and 4 (93%) to Gag. A significantly higher median ELISpot response to Env was seen in high dose recipients (420 SFC/0 6 PBMCs) compared to low dose recipients (57 SFC/0 6 PBMCs), p=0.05. A non-significant difference in response to Gag was seen between groups. All 6 vaccinees (median titer 800, range ) exhibited binding antibodies (Ab) to subtype C gp40 Env but no difference in ELISpot or binding Ab responses was seen between groups after the second HIV-MVA. Neutralizing Ab responses were not detected against pseudovirus 93MW in TZM-bl assay or against SF62 in a PBMC-based assay. Conclusions: This DNA priming-mva boosting strategy is safe and highly immunogenic in Mozambican vaccinees. High dose recipients exhibited higher magnitudes of IFN-γ ELISpot responses to Env than low dose recipients. 28

282 Posters Posters 26: Vaccine Clinical Trials P26.7 T-cell Responses to Novel HIV- Cryptic Epitopes Binghao J. Peng, Jonathan M. Carlson 2, Eric Hunter 3, Anju Bansal, Paul A. Goepfert University of Alabama at Birmingham, Medicine, Birmingham, AL, United States, 2 Microsoft Research, Los Angeles, CA, United States, 3 Emory University, Atlanta, GA, United States P26.8 Topology Influences V2 Epitope Focusing Sergey Shmelkov, Mangala Rao 2, Shixia Wang 3, Michael Seaman 4, Xiangpeng Kong, Shan Lu 3, Timothy Cardozo New York University School of Medicine, New York, NY, United States, 2 Walter Reed Army Institute of Research, Silver Spring, MD, United States, 3 University of Massachusetts Medical School, Worcester, MA, United States, 4 Beth Israel Deaconess Medical Center, Boston, MA, United States Background: Conventional HIV- proteins are thought to be derived from open reading frames (ORF). In addition to ORFs, HIV has the potential to transcribe and translate from five additional reading frames termed ARFs or alternate reading frames. Transcription and translation from ARFs has been shown to occur commonly during HIV, SIV, and other viral infections. As a result of ARF usage, antigenic peptides termed cryptic epitopes (CE) are generated, and becomes a potential source of HLA presented peptides. It is now well established that HIV often mutates to evade T cell recognition. However, many of these escape mutations are detrimental to viral fitness. We therefore hypothesize that CEs that induces viral escape at a population level must be important for controlling HIV. Methods: In order to identify CEs that have escaped T cell recognition at a population level, viral sequences from a Zambian cohort consisting of 40 heterosexual serodiscordant couples were analyzed in addition to each person s HLA allelic information. This revealed multiple HLA-I restricted CEs within gag, nef, and pol regions of HIV-. To validate our finding, 66 cryptic peptides were chosen and used to stimulate cryopreserved PBMC samples from HIV- chronically infected off-art individuals. Results: IFNγ ELISpot assay revealed HIV- CE specific responses at the peptide pool level in 9/32 (28%) individuals tested. However, such responses were not seen in the 6 HIV seronegative donors. We further mapped the positive responders and identified eleven responses at the single peptide level (median magnitude: 33 SFU/0^6 cells; range: SFU/0^6 cells). Additionally, preliminary flow cytometric results revealed CE to also have elicited immune responses in CD4 T cells. Conclusions: Despite their infrequent expression, our data suggests that CEs can enhance breadth of HIV specific T-cell responses. Additionally, since these epitopes were predicted based on HLA associated escape mutations, it further underscores their role in HIV control. Background: The RV44 trial data suggest that a vaccine can prevent HIV infection in humans. VV2 domain s C ß-strand may have been targeted by at least some protective antibodies (Abs) from the RV44 trial. The RV44 trial data suggest that these Abs target the peptide segment from positions 65-8 of the V2 loop (V ) and cross react with all the major HIV subtypes. We hypothesized that we could design a protein immunogen that exclusively presents only this segment in a conformationally appropriate manner and thereby elicit Abs mirroring the protective RV44 Abs. A key question in pursuing this goal is which structural topology of attachment for the VV2 domain s C ß-strand to the immunogen scaffold is more productive for the effective epitope presentation to elicit the desired Abs. Methods: The V peptide was fused to a non-hiv scaffold, cholera toxin B (CTB), in two different structural topologies. First, both ends of the ß-hairpin formed by the B and C strands were directly fused to CTB, representing the topology observed in the VV2 domain. In the second approach, a cyclic permutation of the C and B strands was designed, and then fused to CTB. We tested these designed immunogens by immunization in rabbits using a DNA prime (gp20) - protein boost (CTB-V2) approach. The resulting serum was analyzed for neutralization activity against HIV viruses and for binding of serum Ig to gp20 and gp70-scaffolded VV2 domains from six HIV subtypes. Results: We found that immune sera from the permuted topology, but not the obvious native topology, bind cyclic V2 peptide with high endpoint titers, neutralize Tier HIV viruses from subtypes B, C and AG, and bind gp70-vv2 representing six HIV subtypes. Conclusions: Our data demonstrate that the structural topology influences the presentation of the C-strand for elicitation of Abs. Surprisingly, the non-native topology proved to be more productive: Abs elicited from this design exhibit the same properties as the protective RV44 Abs. 282 HIV Research for Prevention 204 HIV R4P

283 Posters 26: Vaccine Clinical Trials Wednesday, 29 October P26.9 B-cells that Bind HIV Particles Encode CD4- induced, C-like and V3-specific Antibodies that Mediate Broad ADCC Activity Katherine L. Williams, Valerie Cortez,2, Adam S. Dingens 2, Stephanie Rainwater, Julie F. Weis, Xuemin Chen 3, Paul W. Spearman 3, Julie Overbaugh Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 Program in Molecular and Cell Biology, University of Washington, Seattle, WA, United States, 3 Emory University Department of Pediatrics and Children s Healthcare of Atlanta, Atlanta, GA, United States Background: A more complete understanding of the epitopes targeted by an individual s HIV-specific neutralizing and non-neutralizing antibody repertoire will be essential to informing rational vaccine design. Methods: We employed fluorescently-labeled viral like particles (VLPs) generated with a combination of two clade A envs - one the autologous transmitted virus and another a neutralization sensitive virus (Q46. d) - to sort IgG+ memory B cells obtained 2.5 years following HIV infection in QA255, a patient who developed both a broad neutralizing and ADCC-mediating antibody response early following infection. Results: In total, 50 antibodies (Abs) were produced from 92 sorted B cells that bound VLPs. Twelve Abs demonstrated measurable binding to Q46.d VLP by ELISA. None of the Abs demonstrated neutralizing activity against Tier 2 viruses, although three neutralized Tier A and B viruses. Analysis using the rapid and fluorometric ADCC (RFADCC) assay determined that three clonally unrelated Abs mediated potent ADCC activity against both of the Clade A envs used in the VLP sort as well as envs from additional A, B and C clades. Two of the three Abs recapitulated the majority of the heterologous, cross-clade ADCC breadth observed with contemporaneous QA255 plasma. Experiments using Fabs to block epitope-specific ADCC activity indicated that none of the three antibodies resemble A32, an antibody that is directed against a CD4-induced (CD4i) epitope and is typically a dominant response in naturally infected individuals. Additional Fab blocking experiments using CD4i Abs C and 7B, which target distinct epitopes, suggest that the two broadly ADCC-mediating Abs target the C-like epitope. The third Ab, which showed more limited breadth, targets a V3-specific linear epitope similar to Ab 9B. Conclusions: These data demonstrate that a sorting strategy incorporating VLPs can be used to identify both binding and ADCCmediating antibodies that target CD4-induced and variable loop epitopes

284 Posters Posters 27: Adjuvants P27.0 P27.02 Monocyte-derived DC Electroporated with mrnas Encoding Both Specific HIV Antigens and DC Adjuvants Are Able to Improve T-cell Functionality Alberto C. Guardo, Laia Miralles, Joeri L. Aerts 2, Kris Thielemans 2,3, Beatriz Mothe 4, Javier Martinez-Picado 4, Christian Brander 4, Felipe Garcia, Montserrat Plana, ihivarna Consortium IDIBAPS, Retrovirology and Viral Immunopathology Lab. Hospital Clinic, University of Barcelona, Barcelona, Spain, 2 Laboratory of Molecular and Cellular Therapy, Medical School of the Vrije Universiteit Brussels, Department of Physiology-Immunology, Brussels, Belgium, 3 etherna, Kortenberg, Belgium, 4 AIDS Research Institute IrsiCaixa- HIVACAT, Hospital Universitari, Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona, Spain Background: In the context of therapeutic vaccination of HIV-infected patients, we have tested in vitro a combination of mrna sequences that fulfil two main objectives. On the one hand, a specific T cell activation immunogen mrna that focuses the response onto the most vulnerable targets in the HIV viral proteome and on the other hand, a previously tested stimulus (TriMix: a mixture of CD70+CD40L+caTLR4 mrnas) for appropriate activation of antigen presenting cells (DCs). Methods: DCs were generated from peripheral blood monocytes (MDDC) from chronically HIV infected patients by incubation with GM- CSF and IL-4. These cells were electroporated with TriMix (5 µg) and/ or HIVACAT (20 µg) mrna, with their respective controls. After that, DCs were cocultured with autologous PBMCs for up to 6 days. In addition, the maturation profile of MDDCs (CD80, CD83, CD86, CCR7) was analyzed by FACS 24h after electroporation. Functional analysis was performed using different techniques: 25-multiplex Luminex assay, T cell proliferation by CFSE and IFN-γ ELISPOT at different time points. Results: Increased expression of CD80, CD83 and CCR7 was observed on MDDCs upon electroporation with TriMix mrna. Functionally, mrna electroporated MDDCs were able to stimulate T cells from HIV-infected individuals on cart in vitro. In fact, MDDCs electroporated with both HIV antigens and TriMix, induced higher T-cell activation than their respective separated components or whole AT2-inactivated virus in terms of both IFNγ secretion and proliferation. Other Th, Th2 and proinflammatory cytokines showed a similar profile secretion pattern. Finally, a higher proportion of stimulated CD8 + T cells, than of CD4 + T cells, was detected. Conclusions: mrna electroporation of MDDCs improved their maturation status and was able to enhance HIV specific T cells responses. Our results suggest that this mrna combination could be considered for a HIV therapeutic vaccination approach. Intradermal Vaccination against SIV Induces the Activation and Migration of Langerhans Cells in Non-human Primates Lucille Adam,2,3, Biliana Todorova,2,3, Roger Le Grand,2,3, Catherine Chapon,2,3, Frederic Martinon,2,3 CEA, Division of Immuno-Virology, IDMIT Center, Fontenay aux Roses, France, 2 Université Paris-Sud, UMR E, Orsay, France, 3 Vaccine Research Institute (VRI), Créteil, France Background: Modern approaches of vaccination aim at targeting dendritic cells (DC) with specific ligands and adjuvants. Electroporation (EP), which was used to increase the expression of anti-siv DNA vaccine encoded antigens delivered in the skin, strongly improved the specific immune response in non-human primates (NHP). The aim of this work was to study the role of local DC, in the induction of this particular immune response. Methods: auxogtu-multisiv DNA plasmid or PBS was injected intradermally and associated with EP at vaccinated sites in NHP. Skin biopsies of injection sites were performed at 24h, 72h and 8 days after treatment for histology or cell extraction and flow cytometry analysis. Skin DC, stained with an AlexaFluor 488 labeled anti-hla-dr antibody, were imaged with noninvasive in vivo fibered confocal fluorescence microscopy or monitored continuously during 24h by time-laps confocal videomicroscopy on skin explants. Results: EP associated to intradermal injection of the DNA vaccine induced a recruitment of granulocytes and inflammatory monocytes/ macrophages in epidermis and dermis, as well as a population of inflammatory dendritic epithelial cells. In epidermis, 24h after treatment, we observed an initial increase of Langerhans cells (LC) with an upregulation of HLA-DR, CD86 and CD83, demonstrating their maturation, followed by a decrease of LC number, suggesting a migration to draining lymph node. In vivo monitoring confirmed the cell mobilization and showed an increase of DC velocity and displacement after EP. The skin microenvironment analysis revealed a release of pro-inflammatory soluble factors (MCP-, IL-8, IL-5, IL-8) and anti-inflammatory mediators (ILRA and scd40l) by 24h, all considerably enhanced in the presence of the anti-siv vaccine. Conclusions: This study highlights new elements of cell activation mechanisms in the skin and shows that EP induced a local inflammation that led to the activation and mobilization of LC. This opens up new possibilities for vaccine strategies. 284 HIV Research for Prevention 204 HIV R4P

285 Posters 27: Adjuvants P27.03 Combinations of TLR4 and TLR7/8 Adjuvants Administered via the ID or IN Routes Generate Different Vaccine Antigen-specific Immune Outcomes in Minipigs Thursday, 30 October P27.04 Chimeric Nod2/TLR2 Ligand Amplifies HIV- Gag p24-specific Mucosal and Systemic Immune Responses after Sub Cutaneous Immunization in Mice Paul F. McKay, Deborah F. L King, Jamie F. S Mann, Guillermo Barinaga, Darrick Carter 2, Robin J. Shattock Imperial College, Department of Infectious Diseases, Division of Medicine, London, United Kingdom, 2 Infectious Disease Research Institute, Seattle, WA, United States Background: Previous studies have demonstrated synergy between TLR4 and TLR7/8 stimulation with enhanced CD4 T cell cytokine production that may augment CD4 dependent Ab generation. In minipigs we assessed the immune outcome using optimal agonist TLR4/7/8 ligand adjuvant dosage or combination in a mixed ID and IN route vaccination regimen. Methods: Groups of pigs received CN54gp40 via the ID or IN route with optimally titrated R848 or GLA adjuvants. A schedule of 3xIN+2xID was compared to 3xID+2xIN, each inoculation given 3 wks apart. Control animals were unadjuvanted. Pigs were sampled weekly and Agspecific humoral responses assessed by ELISA, avidity and neut assays. Results: Combinations of R848 and GLA had an additive enhancing effect when used ID but surprisingly GLA abrogated R848-induced immunity after IN inoculation. Therefore, ID injections included both R848 and GLA, IN only R848. Optimized adjuvant dose/combinations significantly enhanced Ag-specific responses when administered via either route. At the end of each regimen both groups exhibited similar quantities but qualitatively different serum Ag-specific Ab. The ID route vaccinations elicited sera of significantly higher avidity (p=0.042, week 5-2 weeks post 2 nd vaccination), viral neutralising capacity and mucosal IgG (p=0.006, week 5). Conversely, serum and mucosal IgA responses, although low, were significantly higher in the 3xIN primed group (p< 0.05, week 5). Conclusions: These data begin to address important issues relating to adjuvant combinations and routes of administration. TLR4 and TLR7/8 agonists combined to enhance Ag-driven immune responses but only after ID vaccination. The inhibitory effect of GLA on R848-driven responses after IN inoculation suggests an active contribution by GLA rather than a simple formulation issue, as adjuvant co-formulation did not inhibit R848 responses after ID vaccination. Further investigation of adjuvant combinations in responsive animal models will aid development of effective vaccines. Capucine Phelip, Vincent Pavot, Nicolas Rochereau 2, Eric Perouzel 3, Thierry Lioux 3, Gérard Tiraby 2, Charlotte Primard 4, Stéphane Paul 2, Bernard Verrier Institut de Biologie et Chimie des Protéines, LBTI, UMR 5305 CNRS/ Université de Lyon, Lyon, France, 2 Groupe Immunité des Muqueuses et Agents Pathogènes - INSERM CIC 408 Vaccinologie, Faculté de Médecine de Saint-Etienne, Saint-Etienne, France, 3 CAYLA - InvivoGen, Toulouse, France, 4 ADJUVATIS, Lyon, France Background: Toll-like receptor (TLR) ligands are critical activators of innate immunity and are being developed as vaccine adjuvants. However, their potential synergistic effect in conjunction with other Pattern Recognition Receptor (PRR), like Nod-like receptor (NLR) agonists remains poorly studied. In this study, we evaluated both in vitro and in vivo the adjuvant role of a chimeric Nod2/TLR2 ligand when co delivered with poly(lactic acid) (PLA) biodegradable nanocarriers, loaded with HIV- Gag p24. Methods: Chimeric Nod2/TLR2 ligands have been chemically synthesized and compared with their separate counterparts, Nod2 and TLR2 with regards to their capacities to induce maturation and proinflammatory cytokines of Human Monocytes derived Dendritic cells. After sub cutaneous administration of each ligand, co-delivered with PLA-p24 particles, either alone or in combination, p24-specific Th/Th2 responses have been evaluated in balb-c mice both at systemic and mucosal levels (IgG, IgA, Elispot analysis). Results: In vitro chimeric Nod2/TLR2 ligand induced synergic and strong up-regulation of maturation markers (CD80, CD83, CD86, DC-LAMP), costimulatory molecules at the DCs surface and proinflammatory cytokines secretions (IL- beta, IL-6, IFN alpha, IFN gamma, TNF alpha) compared to separate ligands. Interestingly enough, sub-cutaneous administration of PLA nanoparticles carrying Gag p24 co-administered with Nod2/TLR2 chimera induced strong Ag-specific IgA and IgG antibody titers both at systemic and mucosal sites as strong as Cholera Toxin positive controls. Conclusions: These data demonstrate that targeting both TLR and Nod pathways with a chimeric ligand such as Nod2/TLR2 could induce strong mucosal immune responses after sub cutaneous administration when co-delivered with PLA biodegradable particulate vector. These results point out the interest of this new class of chimeric molecules as HIV vaccine adjuvants to induce both strong systemic and mucosal immune responses after parenteral administration

286 Posters Posters 27: Adjuvants P27.05 Antigen Formulations and Routes of Immunization Drive the Magnitude and Quality of HIV-specific Mucosal Immune Responses in Mice Thomas Vazquez, Léa Torrieri-Dramard, Fabien Pitoiset, James Vigneron, David Klatzmann, Bertrand Bellier I3 Laboratory - INSERM U GHPS Paris, Paris, France P27.06 Alarmin IL-33 Acts as an Immunoadjuvant to Enhance Antigen-specific anti-viral Immunity Daniel Villarreal, Megan Wise, Jewel Walters, Emma Reushel, Min Joung Choi 2, Nyamekye Obeng-Adjei, Jian Yan 3, Matthew Marrow 3, Niranjan Sardesai 3, David Weiner University of Pennsylvania, Philadelphia, PA, United States, 2 Korea Food and Drug Administration, Osong-eup, Korea, Democratic People`s Republic of, 3 Inovio Pharmaceuticals, Inc., Blue Bell, PA, United States Background: Despite considerable efforts, the development of a successful HIV vaccine remains challenging. Current attention is mainly focused on developing strategies to enhance the immunogenicity of vaccine formulations and the induction of mucosal immunity. Methods: Here, we investigated the influence of particulate form of antigen and the route of administration in the induction of HIVspecific immune responses and mucosal immunity. We evaluated the immunogenicity of HIV-gp40 antigen expressed as proteins or recombinant Virus-Like Particles (VLPs) and compared different mucosal (nasal (IN), rectal (IRec), vaginal (IVag)) and parenteral (intradermal (ID), subcutaneous (SC)) routes of immunization, or their association in primeboost strategies. Results: We show that the antigen formulation and route of immunization are critical factors governing the vaccine efficacy. We observed that DNA vaccine that express HIV-gp40 on VLPs induced higher levels of immune responses than standard DNA expressing non-particulate antigens, especially at mucosal level after IN DNA administration. Similarly, we observed that VLP boost, when administrated by IN or IVag routes, improved T cell multifunctionality at systemic and mucosal levels as compared to protein boost. When mucosal and parenteral immunization were compared, we show that IgA and IgA-secreting cells were only induced in mucosal tissues when DNA priming was done by IN route, but not by ID. We also compared IVag and IRec VLP boosts after IN DNA primes and demonstrated that both increased antigen-specific T- and B-cell responses in systemic and local compartments, inducing HIV-specific IgA-secreting cells at proximal mucosa to the site of immunization. Conclusions: Taken together, our results showed the importance of the particulate formulation of the antigen, such as VLPs, and the role and the interest of mucosal routes of immunization to improve local immune responses. Background: DNA vaccines are an important approach for the generation of humoral and cellular immune responses in vivo. We have recently reported that combining an IL-2 expression vector along with HIV DNA encoded antigens can induce robust T cell responses in the clinic (HVTN 080). Identification of novel adjuvants at the innate to adaptive immune border will likely provide further improvements in DNA vaccine potency. In this regard we have recently developed a novel alarmin IL-33 as a new immunoadjuvant in the synthetic EP delivered DNA vaccine platform. We hypothesized that this endothelial cell alarmin would be in a particularly effective position to transition the adaptive immune response after viral infection. Methods: Mice were immunized with immune plasmid adjuvant IL33 in combination with DNA followed by EP two to three times at three week intervals.one week post final vaccination, spenocytes were harvested to investigate cellular responses. Results: We report that IL-33 is capable of enhancing potent antigen (Ag)-specific effector and has a similar robust effect on enhancing memory T cell immunity in vivo in a DNA vaccine setting. Additionally, IL-33 augmented vaccine-induced Ag-specific polyfunctional CD4+ and CD8+ T cell responses. Importantly a large proportion of CD8+ T cells undergo cytolytic plurifunctional degranulation that is improved over DNA vaccine alone. We show that IL-33 can particularly expand the magnitude of Ag-specific CD8+ T cell responses and elicit stronger and longer lived effector-memory CD8+ T cells. For HIV antigens, both the magnitude as well as the phenotype of T cells induced by the IL-33 DNA vaccine are improved. Conclusions: We have now developed a Rhesus IL-33 genetic adjuvant for studies in NHP which allow for more detailed studies including challenges to be performed. These studies will be informative for HIV vaccine development and lay the groundwork for this novel genetic adjuvant for study in the HIV vaccine setting. 286 HIV Research for Prevention 204 HIV R4P

287 Posters 28: Behavioral and Social Sciences Thursday, 30 October P28.0 Does a Weighted Analysis Make any Difference in the Estimates from a Sample Survey? Rajatashuvra Adhikary, Prabudhyagopal Goswami 2, Mandar Mainkar 3 FHI 360, Washington DC, DC, United States, 2 FHI 360, New Delhi, India, 3 National AIDS Research Institute, Pune, India Background: Self-weighted sampling designs are not always feasible in sample surveys. Sample weights are calculated and applied to have unbiased estimates. However, it is important to understand the extent of difference between weighted and un-weighted estimates. We examined these differences among men who have sex with men (MSM) participants using data from an integrated bio-behavioral survey conducted in 2009 in four districts of Andhra Pradesh, India. Methods: Two-stage time location cluster sampling approach was used to recruit,608 (around 400 from each district) men aged >8 years who had sex with another male in one month prior to survey. Sample size was calculated based on 95% confidence level, 90% power, and design effect =.7. Consented participants provided behavioural data in structured questionnaires and blood and urine specimens. Specimens were tested for HIV and syphilis. Multivariate analysis was performed to determine correlates of CCU and HIV prevalence adjusted for sociodemographic variables. Results: Weighted and un-weighted estimates of consistent condom use (CCU) with regular male partners, paying male partners, and casual male partners, HIV prevalence, and syphilis prevalence varied across the four districts. The absolute difference between the weighted and un-weighted estimates ranged from 0.7% to 3.2% for CCU and from 0.2% to 5.3% for HIV and syphilis. Z-scores show a significant difference between weighted and un-weighted estimates in many cases. Weighted and un-weighted multivariate analysis shows opposite trends in the adjusted odds ratio (AOR) in few cases and relatively large differences in the significance level (p-value). Conclusions: Weighted analysis for a probability-based sample survey provides unbiased population estimates whereas un-weighted estimates are representative of the sample. If possible, a self-weighted sampling design should be adopted, allowing for unbiased estimation for the sample and population

288 Posters Posters 29: Circumcision and Acceptability P29.0 A Qualitative Exploration of Medical Male Circumcision among Young Men from Whizzkids United, Edendale, Pietermaritzburg Rusha Govender, Janan Dietrich 2, Jenny Coetzee 2, Marcus Mc Gilvray, Douglas Wilson Edendale Hospital, Pietermaritzburg, South Africa, 2 PHRU, Johannesburg, South Africa P29.02 Assessing the PrePex Device for Voluntary Medical Male Circumcision in South Africa Limakatso Lebina, Noah Taraburekera 2, Minja Milovanovic, Nkeko Constance Tshabangu, Neil Martinson Perinatal and HIV Research Unit, Johannesburg, South Africa, 2 Population Services International, Represented in South Africa by Society for Family Health, Johannesburg, South Africa Background: In Africa male medical circumcision (MMC) has been shown to reduce HIV acquisition by up to 60%. WHO/UNAIDS recommendations emphasize that male circumcision should be considered an effective intervention for HIV prevention in countries and regions with heterosexual epidemics, high HIV and low male circumcision prevalence. Mathematical models based on the results of randomized controlled trials suggest that medical circumcision could avert two million HIV infections over the next 0 years; however it is unclear whether medical circumcision would decrease the overall prevalence of HIV infection when delivered outside the context of the controlled environment of a randomized trial. This study was conducted at WhizzKids United (WKU) based at Edendale Hospital, Kwa-Zulu Natal. WKU is a grassroots South African nonprofit organization that aims to empower and inform high risk youth about health care and HIV/AIDS prevention. The MMC service facility at Edendale Hospital performs around 200 circumcisions a month. WhizzKids United has made the uptake of MMC services more accessible for adolescents. Methods: Thirty-one in-depth Interviews and eight focus group discussions (FGDs) were conducted with young males aged 4-24 years. Fifty percent of the young males were uncircumcised and fifty percent reported being circumcised. There were two focus group discussions per age group were held. One of these consisted of circumcised individuals and the other consisted of uncircumcised individuals. Thematic analysis was used for data analysis. Results: The global themes identified from the uncircumcised participants included, () medical, (2) peer pressure, (3) decision-making, (4) sexual risk-taking behaviours. The global themes identified for circumcised participants included, () medical, (2) traditional, (3) Becoming a man and (4) sexual risk- taking behaviours. Conclusions: The overall findings suggest that young men in Kwa-zulu Natal are accepting of medical male circumcision. Background: Voluntary medical male circumcision has been proven to reduce the risk of acquiring HIV by at least 60% in men who are circumcised. South Africa is scaling-up medical male circumcision, and the number of circumcised men is increasing. However, if circumcision could be safely simplified without compromising efficacy, it could potentially allow for more circumcisions to be done. The PrePex device was developed to simplify the circumcision procedure by making sutures, diathermy, scalpels, and local anesthesia unnecessary. However, there is no data available on the assessment of the PrePex device in South Africa. Methods: A phased, multisite, non-randomised study in which adolescents (4-7 years) and adult (8-45) men underwent circumcision using the PrePex device. Data were collected from 9 visits; application, 2 telephonic follow-ups, removal and 5 follow-up visits. Outcome measures include adverse events, pain and discomfort, procedure time, sexual resumption, healing time, and PrePex sizes. Data presented is of the first 38 participants. Results: A total of 264 adult men and 54 adolescents were recruited for the PrePex study across the three sites. The overall moderate and severe adverse events (AE) rate was 2.5% with none of the adverse events requiring hospitalisation. Mild AE were predominantly obstructed urine flow. The most commonly used PrePex sizes were size B and C (63%, 97/32). The median resumption for sexual activity was 44 days (IQR: ). The device application procedure was quick and caused minor to no pain however the removal procedure was more painful but the pain was temporary. Conclusions: The preliminary results indicate that the PrePex procedure was easily learnt by the staff at the three sites. The device application is quick, safe and less painful however removal causes temporary pain. The AEs encountered were similar in severity to the blade based circumcision with the exception of urinary obstruction. 288 HIV Research for Prevention 204 HIV R4P

289 Posters 29: Circumcision and Acceptability P29.03 Assessing the Acceptability of the PrePex Device for Voluntary Medical Male Circumcision in South Africa Minja Milovanovic, Noah Tarabureka 2, Nkeko Constance Tshabangu, Mmatsie Manentsa, Neil Martinson, Limakatso Lebina Perinatal and HIV Research Unit, Johannesburg, South Africa, 2 Population Services International, Represented in South Africa by Society for Family Health, Johannesburg, South Africa Thursday, 30 October P29.04 LB Does Circumcision Offer Some Protection from HIV for Men who Have Sex with Men? A Cross-sectional Study in China Han-Zhu Qian, Yuhua Ruan 2, Yu Liu, Douglas F. Milam, Hans M.L. Spiegel 3, Lu Yin, Dongliang Li 4, Yiming Shao 2, Sten H. Vermund Vanderbilt University, Institute for Global Health, Nashville, TN, United States, 2 China CDC, State Key Laboratory for Infectious Disease Prevention and Control, NCAIDS, Beijing, China, 3 NIAID, NIH, Henry M. Jackson Foundation - Division of AIDS, Bethesda, MD, United States, 4 Chaoyang District CDC, Beijing, China Background: Following the recommendation by the World Health Organisation, medical male circumcision is being scaling-up in South Africa. Circumcision reduces the risk of acquiring HIV by 60%. Most circumcision facilities in South Africa use the surgical procedure with very little use of devices. Previous studies have shown that PrePex is a quick, easy and acceptable method of circumcision. However, there is no information on the acceptability of the PrePex device in South Africa. Methods: A phased, multisite, non-randomised study in which different individuals across sites in South Africa, were approached to answer a structured acceptability questionnaire: PrePex users, people accompanying men coming for circumcision, men refusing PrePex, and healthcare workers who worked with PrePex and those that would work with it if it were rolled out. A total of 6 questionnaires were used to explore: cultural acceptability, trust, recommendation of PrePex and perceived advantages and disadvantages. Results: A total of 439 participants, across the different groups, were recruited for the PrePex acceptability study. The PrePex device was considered culturally acceptable by 86% of PrePex users, 60% of the men who refused PrePex and 67% of accompanying. The device was also highly trusted by PrePex users (99%), accompanying persons (95%) and 55% of refusals indicated they trusted it. When asked to explain majority of the responses include perceived safety compared to the blade procedure. Overall, there was acceptance of the PrePex device and procedure by healthcare workers, with many respondents expressing that it is an alternative to traditional circumcision. Furthermore, there was a strong expression that if nurses were to do PrePex there should be a certificate from a relevant body. Conclusions: The results indicate that the PrePex device and procedure is considered relatively acceptable and there appear to be no major social barrier to implementation however, cultural differences were noted. Background: The observational literature on the association of male circumcision and odds of HIV among men who have sex with men (MSM) is often limited by a failure to definitively ascertain circumcision status and/or by a failure to carefully assess sexual position preferences (insertive, mixed/versatile, or receptive). No randomized clinical trials of circumcision have been conducted among MSM. We conducted a crosssectional study of MSM in Beijing, China to assess associations, if any, of circumcision and HIV, controlling for anal sexual positioning role. Methods: MSM were recruited both from the community and from HIV clinics. Circumcision status was evaluated by genital examination by staff trained by an expert urologist. Anal sexual role was assessed by questionnaire interview. Associations of circumcision status with HIV infection were assessed through multivariate logistic regression analysis. Results: The odds of HIV infection by circumcision status and preferred anal sexual role in 053 MSM were as follows: Receptive/versatile/uncircumcised (n=54, 77 HIV+/ 364HIV-) REFERENCE GROUP Receptive/versatile/circumcised (n=47, 9 HIV+/ 38HIV-) aor=0.48 (0.22,.02) Insertive/uncircumcised (n=434, 76 HIV+/ 358HIV-) REFERENCE GROUP Insertive/circumcised (n=3, 2 HIV+/ 29HIV-) aor=0.35 (0.3,.0) The OR was adjusted for age, ethnicity, marriage, education, occupation, and Beijing vs. non-beijing legal residency. Conclusions: Circumcised MSM were less likely to have acquired HIV, especially among men practicing predominantly insertive anal intercourse, with effect sizes comparable to those seen for heterosexual men in Africa. However, findings may have been due to chance, given upper bound 95% CI for aor >.0. Circumcision needs clinical trials evaluation as a potential global HIV prevention strategy for MSM. The HIV Prevention Trials Network is developing a feasibility study (HPTN 079) for such a trial

290 Posters Posters 29: Circumcision and Acceptability P29.05 LB HIV Is the Primary Exclusion Criterion in a PrePex Male Circumcision Device Introductory Study in Mozambique P29.06 LB Women s (Mis) Understanding of Male Circumcision: A Mixed Methods Study in Zambia Mehebub Mahomed, Beverley Cummings 2, Jotamo Come 3, Bossemeyer Debora, Thais Ferreira, Leonel Nhambi, Edgar Necochea 4, Humberto Muquingue, Kelly Curran 4 Jhpiego, Maputo, Mozambique, 2 Centers of Disease Control and Prevention/Mozambique, Maputo, Mozambique, 3 Ministry of Health, National Programme for VMMC Expansion, Maputo, Mozambique, 4 Jhpiego, Baltimore, MD, United States Background: Voluntary medical male circumcision (VMMC) reduces female to male HIV transmission by approximately 60% and is recommended by WHO and UNAIDS as a priority intervention in high HIV prevalence settings. In Mozambique, VMMC for HIV prevention started in 2009; more than 300,000 males were surgically circumcised by March 204; the goal is 2 million by 206. PrePex could potentially reduce procedure time and increase acceptability of VMMC because it does not require injectable anesthesia or suturing. In 203 an introductory study of the PrePex device was conducted in Maputo, Mozambique to assess the acceptability among providers and clients. Methods: Adult clients presenting for VMMC at the study site were offered surgical or PrePex circumcision. Those who preferred PrePex were screened for inclusion criteria. Exclusion criteria were recorded. The current WHO guidelines exclude HIV+ men from device circumcision. Results: During the study, 752 clients aged 8 or older presented for VMMC and were offered the choice of PrePex or conventional surgery; 6 (5.4%) preferred surgical VMMC. Of the 636 clients who chose PrePex, 32 (20.8%) were ineligible. HIV infection was the primary reason for exclusion, restricting 85 (64%) interested HIV+ clients. Phimosis or narrow foreskin was present in 7 (3%) of the ineligible clients. Sixteen clients (2%) were unable to communicate in Portuguese and 8 (6%) lacked communication means (cell phone); both were study requirements. The remaining 6 (5%) were excluded due to active STI, sexual dysfunction or previous penile surgery. Conclusions: One third of adult clients offered PrePex either did not want device circumcision or were ineligible under current guidelines that exclude HIV+ men. An integrated program offering both device and surgical VMMC remains the best service delivery option. However, there is need to assess the safety of PrePex among HIV positive clients, as HIV testing in Mozambique is recommended but not required in routine VMMC service delivery. Nicole A. Haberland, Christine A. Kelly, Drosin M. Mulenga 2, Paul C. Hewett 2, Barbara S. Mensch Population Council, New York, NY, United States, 2 Population Council, Lusaka, Zambia Background: Randomized controlled trials have demonstrated that male circumcision (MC) reduces men s risk of heterosexual acquisition of HIV, but there is no evidence that individual women benefit directly from MC. Women s understanding of the protection afforded - or not - by MC against HIV and STIs has important implications for risk compensation and demand. Methods: This mixed methods study explores the prevalence, depth, and correlates of women s understanding of four dimensions of MC knowledge: awareness of MC, knowledge that MC partially protects males against HIV, knowledge that MC reduces males STI risk, and knowledge that MC has no effect on females HIV risk if she has sex with a man who is circumcised. We combine data from the first two rounds of a longitudinal study of Zambian women aged 5-29 (n=933) with indepth interviews conducted among a subsample of respondents (n=45). Results: Although awareness of MC was high - 77% of women at baseline reported having heard of MC before it was described in the interview - women lacked more nuanced knowledge of MC s protective effects. In both the quantitative and qualitative surveys, a disconcerting proportion of women incorrectly believed that MC fully protects men from HIV and STIs, and that MC similarly offers women partial - or even complete - protection. Multivariable analysis showed that more highly educated and wealthier women were better informed about MC than their more disadvantaged peers. Conclusions: Efforts are needed to increase understanding about the limits of MC protection, particularly among more marginalized women who may have difficulty accessing accurate information about MC or harbor misperceptions about its protective effects. Messaging surrounding MC should also more explicitly address women s needs. Health professionals should emphasize that, because MC provides men with only partial protection against HIV and some STIs, women are still at risk regardless of their partner s circumcision status. Condoms remain critical. 290 HIV Research for Prevention 204 HIV R4P

291 Posters 30: Condoms: Attitudes, Use and How to Increase Thursday, 30 October P30.0 Behavioral Attitudes towards the Use of Male Condoms as a Means of Preventing the HIV/ AIDS Spread Patrick Bitangumutwenzi,2,3, Denis Sinzinkayo 3 Yowli Burundi, Bujumbura, Burundi, 2 University of Burundi, Medicine, Bujumbura, Burundi, 3 Health Healing Network Burundi, Bujumbura, Burundi P30.02 Why Young MSM Do Not Use Condoms Consistently: A Qualitative Exploration Tareerat Chemnasiri, Anchalee Varangrat, Supaporn Chaikummao, Anupong Chitwarakorn 2, Timothy H. Holtz,3 Thailand MOPH - U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Thailand Ministry of Public Health, Department of Disease Control, Nonthaburi, Thailand, 3 Centers for Disease Control and Prevention, Division of HIV/AIDS Prevention, Atlanta, GA, United States Background: Assess knowledge of condom use among adolescents of secondary school, Evaluate the cultural and/or religious impact on the use of condoms in adolescents and Improve the use of condoms as a means to fight against HIV/AIDS, STIs and undesired pregnancies. Methods: The first round consisted of conducting a direct interview towards adolescents with data collection on a questionnaire. The second time, we gave a survey form to fill out and we compared the results of the interview with those of the survey form. Close monitoring helped to yield tangible results. Results: Of 256 adolescents, 23 were Catholics, 6 were Protestants, and 38 were Muslims while 34 were pagans. The extremes of age were 3 and 22 with an average age of The sex ratio M/F was 4/3. Of 256 adolescents, 88 were from poor settings with a precarious level of cultural belief. Of them, 4 agreed that they use condoms regardless of religious and cultural barriers; 48 agreed that they can never use condoms due to the religious Barriers; 67 admitted that the condom does not have the same sexual pleasure and so do not use it. Of 256, 7 have already had genital problems and have consulted the doctor. The study was able to show that most Protestants deny the condoms and preach abstinence; Muslims willingly use condoms while the Catholics are conservative. Conclusions: Religious beliefs affect condom use among adolescents while culture has no great impact. It is necessary to improve sexual practices in the adolescents environment to prevent the spread of HIV/AIDS and other STIs, unwanted pregnancies, etc. and promote educational achievement. Background: High HIV incidence and inconsistent condom use were found among young (8-24 year-old) men who have sex with men (YMSM) in a recent Bangkok cohort study (8/00 person-years). We explored reasons behind inconsistent condom use among YMSM using qualitative methods. Methods: Eight focus group discussions and ten key informant interviews were conducted between June 202 and June 203. Individuals were randomly selected by age and history of risk behaviors from MSM participating in a five-year cohort study in Bangkok, Thailand. We collected socio-demographic and behavioral data using a short questionnaire. Group discussions and interviews were audio-recorded, transcribed, and analyzed using Atlas.ti version 7.7. Results: All 47 participants were Thai MSM, 8 to 24 years of age, who were asked to discuss issues related to inconsistent condom use. Participants expressed feeling an unnatural sensation while using a condom, having a latex allergy, and feeling pain from sexual penetration during anal intercourse which reduced sexual pleasure and sensitivity of penile erection and orgasm. Some participants reported difficulty finding extra-large or extra-small condoms. Fear of rejection by partners and lack of condom negotiation skills were noted by receptive sexual role participants. Individual sexual preference, sensation seeking behavior, and overwhelming sexual desire were perceived to promote non-condom use. Being out of condoms and not carrying a condom at all times were also expressed as promoting inconsistent condom use. Participants believed not using condoms with a steady partner to be a sign of mutual trust. Lack of awareness and misconceptions of HIV transmission caused YMSM to overlook using safe sex practices. Conclusions: HIV awareness, condom negotiation skills, attitude change about condom use, and same-sex education are needed for YMSM in Bangkok. Condom use with steady partners should be strongly emphasized. Variety of condom size and allergy-free condoms are needed for this population. 29

292 Posters Posters 30: Condoms: Attitudes, Use and How to Increase P30.03 P30.04 Exploring the Impact of Social Marketing of Female Condoms in the City of Kumba, Cameroon Alemju Fontu,2 Association Camerouniase pour Marketing Social, Kumba, Cameroon, 2 Forestry Conservation and Environmental Society, Kumba, Cameroon Background: Female condoms are barrier contraceptive methods that provide protection against conception and acquisition of STIs including HIV. These barriers which are used and controlled largely by females are said to provide autonomy and empowerment over other contraceptive choices. Methods: The project implemented the following interventions. Conducted community sensitization campaigns to increase awareness and benefits of using female condoms.recruited and trained volunteer peer educators on interpersonal communication skills, benefits and use of female condoms.peer educators go out twice a week to strategic locations such as the street sides, market places and other public hubs to educate people on the benefits and correct usage of female condoms Results: At the end of these interventions conducted from January 200 to December 20, 58, 650 persons were reached (Youths 60,000 (37.8%), Women 88,400 (55.7%) and Men 0,250 (.6.5%). Correct use of the female condoms (FC2) was demonstrated to an audience of 9,230 [Men 2200 (23%), Women 3,680 (39.8%) and Youths 3,350 (36.2%)]. Eighty sale points were established at strategic locations in each community and 72,000 (FC2) were sold by peer educators. 20,400 and 5,600 females and males respectively bought the (FC2). 90% of the persons interviewed disclose seeing a female condom for the first time during our campaigns and all females who bought the (FC2)promise to adopt the (FC2) as their contraceptive choice. Conclusions: Outcome of these interventions reveal that most females prefer (FC2)to male condoms. The government and other international donors should support the distribution of (FC2)in order to scale up prevention of HIV/AIDS in sub-saharan Africa Changes in Sexual Behaviour and HIV Prevalence among Married Fishermen along Lake Victoria at two Time Points: A Scorecard for Prevention Efforts Zachary Arochi Kwena, Isaac Mwanzo 2, Chris Shisanya 3, Lilian Achiro 4, Norton Sang 4, Elizabeth Bukusi 4 Kenya Medical Research Institute, Center for Microbiology Research, Kisumu, Kenya, 2 Kenyatta University, Community Health, Nairobi, Kenya, 3 Kenyatta University, Geography, Nairobi, Kenya, 4 Kenya Medical Research Institute, Center for Microbiology Research, Nairobi, Kenya Background: There are considerable efforts towards reducing new HIV infections in key affected populations such as fishermen. Assessing changes in sexual behaviour and HIV prevalence is an important feedback to these prevention efforts. We evaluated changes in sexual behaviour and HIV prevalence among married fishermen in fish-landing beaches in Kisumu County, Kenya. Methods: We analyzed data from two surveys conducted in 2005/6 with 64, and 20/2 with 545 married fishermen to evaluate changes in their sexual behaviour and HIV prevalence at two time points. The participating fishermen in both surveys were randomly sampled from all 33 fish-landing beaches in Kisumu County. The numbers sampled from each beach were proportional to the population size of the beaches. In both surveys, we collected data on socio-economic, sexual behaviour and HIV sero-status. Results: A higher proportion of fishermen in 20/2 survey compared to 2005/6 survey reported drinking alcohol before sex with extramarital partners (27.7% versus.4%; p=0.05) and being involved in transactional sex (65.8% versus 25.0%; p< 0.0). However, more fishermen in 20/2 compared to 2005/6 survey used condoms with extra-marital partners (34.2% versus 5.4%). Overall HIV prevalence in 20/2 survey was marginally lower compared to 2005/6 (2.0% versus 28.0%; p=0.07). However, there was significant 5 percentage point drop in HIV prevalence among fishermen below 25 years old that represent recent infections. Conclusions: Despite increases in other high risk sexual behaviours, condom use with extra-marital partners in this HIV key affected population increased explaining significant drop in HIV prevalence among youth who represent recent infections. 292 HIV Research for Prevention 204 HIV R4P

293 Posters 30: Condoms: Attitudes, Use and How to Increase Thursday, 30 October P30.05 P30.06 Repetitive Risk Reduction Counseling on Condom Use among HIV Exposed Seronegative (HESN) Persons in Jos, Nigeria Evaezi Okpokoro, Sophia Osawe,2, Ruth Daitiri 2, Grace Choji 2, Stephen Umaru 2, Felicia Okolo 2, Pam Datong 2, Alash le Abimiku,2,3 Institute of Human Virology, Abuja, Nigeria, 2 Plateau State Human Virology Research Centre, Jos, Nigeria, 3 Institute of Human Virology, University of Maryland, School of Medicine, Baltimore, MD, United States Background: Condom is the commonest HIV preventive strategy. Over time, the successes of this strategy have been challenged with several socio-cultural factors such as male dominance over sexual negotiations; feeling of mistrust among spouses over condom use and wanting to achieve pregnancy especially in the African continent. We document here condom use during a 2-year follow up among HIV exposed seronegatives in a discordant relationship after consecutive risk reduction counseling in preparation for a future HIV prevention trial in Nigeria Methods: We conducted a prospective cohort study and followed up 534 HESN partners in established sero-discordant relationship (i.e. at least 3 months). Relevant ethical approvals and informed consent were obtained. We provided risk reduction counseling for 0-2 minutes with emphasis on: importance and proper use of condoms along with free condoms; the need to watch for symptom of STIs and request immediate treatment; and the benefit of their HIV+ partner attaining viral suppression and elevated CD4 count before achieving pregnancy and thereafter administered standardized questionnaires on risk behavior. Clinical examinations were done and samples collected for rapid HIV test and safety labs Results: 534 enrollees were eligible for 0 follow-up visits with a mean age of 36 years (9-65years). 257 (48.%) were female and 277 (5.9%) males. A total of 7 individual based risk reduction counseling sessions were provided. About 60% of our female participants are within the child bearing age (i.e. 9-35years) which explains why these are aspiring to achieve pregnancy. More so, this group accounts for the 52% who inconsistently use condom. Nonetheless condom use increased from 40% at baseline to 48% at visit 7 with only 5.8% of females achieving pregnancy Conclusions: Among HESN in a marital relationship, repetitive risk reduction counseling improved the use of condom which highlights the need for combine HIV preventive strategies considering their significant exposure to HIV Acceptability and Ease of Use of New Female Condom Designs among Women Attending an Urban Reproductive Health Clinic in Durban, South Africa Mags E. Beksinska, Jennifer A. Smit, Greener Ross, Busi V. Maphumulo, Nonhlanhla Mphili, Sthe Chonco MatCH Research, Obstetrics and Gynaecology, Durban, South Africa Background: The availability of new female condom (FC) designs, which may improve FC acceptability and affordability, will increase options for couples who choose FCs as their contraceptive and/or disease prevention method. The acceptability of 2 new products: HLL FC and Cupid 2 (smaller version of the existing Cupid FC) compared to the currently available FC2 was evaluated as part of a trial assessing the functional performance of the 2 new FCs. Here we present the acceptability results of the trial. Methods: This randomized, comparative cross-over clinical trial of 3 FCs was conducted among 300 women in an urban reproductive health clinic in Durban, South Africa. Interviewer- assisted surveys were employed during 3 follow-up visits to gather data on comparative acceptability. Numbers and percentages of women in each category of acceptability were calculated. Results: In total, 277 women (92.3%) have completed the study to date using all 3 FC types. Mean age was 27.3 years and 22% had previously used FCs. Of the total sample entered and analysed to date (n=3), over 80% of women liked all FC types very much or liked them somewhat and over 90% said each type was comfortable to use For individual features there were minimal differences between the condoms however two-thirds of women (66.0%) liked very much the pink colour of Cupid2 compared to 53% of the FC2 and HLL FCs. Although over a third of women 38.9% found any FC type difficult to insert but improved with practice at first follow-up, by the last follow-up visit only a fifth (9.5%) reported this, with very easy to insert any FC increasing from 9.7% to 37.2% by last visit. Conclusions: Results from this study show that regardless of FC type, ease of use increased across each follow-up visit. Although women expressed preferences for different FC features, acceptability of all FCs overall was high. A greater range of FCs will provide women with more choice of protection. Data for the full sample of 300 women completing the study will be presented

294 Posters Posters 3: Drug Transporters P3.0 Characterisation of Drug Transporter Gene Expression in Colorectal Tissue and Cell Lines: Induction with Anti-retrovirals for Microbicide Optimization Indrani Mukhopadhya, Susan Berry, Emad M. El-Omar, John Thomson, Georgina L. Hold, Karolin Hijazi University of Aberdeen, Aberdeen, United Kingdom P3.02 Expression of Drug Transporters in Cervicovaginal Cell Lines and Modulatory Effect of Candidate Anti-retroviral Microbicides Kieron A. Smith, Indrani Mukhopadhya, Susan Berry, Georgina L. Hold, Karolin Hijazi University of Aberdeen, School of Medicine and Dentistry, Aberdeen, United Kingdom Background: Drug transporter expression in the colorectal epithelium is likely to play a role in the mucosal disposition of anti-retrovirals (ARVs) in rectal microbicidal preparations and impact their efficacy in prevention of HIV- infection. However, there is limited information on expression levels available. This study assessed expression of 84 drug transporter genes in human colorectal tissue and representative cell lines pre and post ARV exposure. Methods: Drug transporter mrna expression was quantified from colorectal biopsies (n=2) and 6 colorectal cell lines using real time PCR. Relative mrna expression was quantified in CaCo-2 cells after induction with tenofovir (TFV; 000 µm) and dapivirine (DPV; 0 µm) for up to 3 days. Data was analysed using Pearson s correlation (r), hierarchical clustering and principal component analysis. Results: Fifty-eight of the 84 transporters were expressed in colorectal tissue. SLC28A2/CNT2 was the most expressed uptake transporter (>25 fold increase) followed by efflux transporters ABCB/P-gp and ABCC3/ MRP3 (4-5 fold increase). No difference was noted between individual patients, either sexes or biopsy sites (rectum or sigmoid) (r= ). Similarities between tissue and cell lines were low (r values ). Principal component analysis showed distinct clustering of colorectal tissue and cell lines. TFV stimulation of CaCo-2 cells resulted in >5 fold increase in expression of SLC28A3, SLC7A8 and TAP. DPV stimulation resulted in >6 fold increase in expression of SLC7A (26 fold), TAP (2 fold) SLC3A2, SLC38A2, SLC6A3,and ABCA3. Conclusions: This study has enumerated drug transporter mrna expression in colorectal tissue and cell lines. There was partial correlation between cell lines and tissue limiting cell line use as in vivo surrogate. TFV and DPV did not induce efflux transporters in Caco-2 cells, which could result in enhanced drug delivery to submucosal CD4 T cells. Further studies will elucidate whether combination of ARVs will be similarly effective. Background: Drug transporters expressed in the cervicovaginal (CV) epithelium are likely to influence delivery of antiretroviral (ARV)-vaginal microbicides to subepithelial target cells for HIV-. This study aims to characterise drug transporters in CV cell lines and investigate the impact of dapivirine (DPV) and darunavir (DRV) on gene expression for development of in vitro assays for testing transport of candidate ARVmicrobicides across the CV epithelium. Methods: Expression of 84 human drug transporter genes was investigated in HECA, EndE6E7, EctE6E7 and VK2E6E7 using RTqPCR. The impact of DPV and DRV on HECA and VK2E6E7 drug transporters expression was analysed over 72 hours. Results: EndE6E7, EctE6E7 and VK2E6E7 cell lines showed similar baseline expression profiles distinct from HEC-A. ARV-associated uptake transporters ENT, ENT2, OATPD, and OATPE were expressed in all cell lines. OATP8 was expressed in HECA only, and CNT3 expressed in all but HECA. Expression of ARV-efflux transporters P-gp and BCRP was low across all cell lines. Upon stimulation of HEC-A with DPV (0uM) upregulation of efflux transporters MRP2, MRP5 and downregulation of uptake transporters OATP8, OATPE was observed. VK2E6E7 stimulation with DPV showed downregulation of OATPE and CNT3. DRV (250uM) induced expression in HEC-A of MRP2, MRP5, MRP7 and P-gp. VK2E6E7 stimulation with DRV resulted in upregulation of OATPD, CNT3, MRP3 and MRP5 and downregulation of MRP4, MRP7 and OATPE. All reported expression changes following ARV stimulation were > 2 fold. Conclusions: In the absence of ARVs the cell lines investigated express uptake transporters reported in CV tissue, but minimal levels of ARVefflux transporters P-gp and BCRP. Cell lines transfected to overexpress P-gp and BCRP may be suitable for use in transport studies. The downregulation of uptake transporters and upregulation of efflux transporters induced by both ARVs, seen in HECA, may suggest reduced drug delivery to subepithelial target cells in the CV tract. 294 HIV Research for Prevention 204 HIV R4P

295 Posters 3: Drug Transporters Thursday, 30 October P3.03 Transport Characteristics of Antiretroviral Drugs - Single Agents, Double and Triple Combinations in Caco-2 Cells Magda Swedrowska, Abhinav Kumar, Charles Kelly 2, Ben Forbes King s College London, Institute of Pharmaceutical Science, London, United Kingdom, 2 King s College London, Oral Immunology, London, United Kingdom Background: Combinations of antiretroviral (ARV) drugs, Tenofovir (TFV), Dapivirine (DPV) and Darunavir (DRV), provide enhanced antiviral activity compared to single agents when tested in vitro. However, the impact of co-formulation on drug absorption after delivery to the colorectal mucosa is unclear, and methods are required to screen drug-drug interactions. The aim of this study is to investigate the permeability of TFV, DPV and DRV across the colo-rectal epithelium, any role of transporters in regulating absorption and assess the effect of coadministering the microbicidal agents on their transport. Methods: Permeability of TFV, DPV and DRV was investigated in vitro using the Caco-2 epithelial cell model grown on Transwell inserts for 2-28 days. The permeability of ARV drugs was measured, interactions with transporter and effect of co-administration of these ARV drugs, were evaluated. Results: The respective absorptive and secretory permeability of TFV across Caco-2 cell monolayer was 0.2± cm/s and 0.3± cm/s. DPV demonstrated high permeability coefficient, 36± cm/s and 27± cm/s in the absorptive and secretory directions, respectively. TFV and DPV flux was not influenced by the presence of transporter inhibitor verapamil or co-administration with other ARV. Transepithelial transport of DRV was 6-fold greater in the secretory direction compared to absorptive direction. Modulation of the transport of DRV at 0 µm by verapamil was shown (ER decreased from 6 to ). Co-administration of DRV with TFV, DPV and DPV/TFV in combinations resulted in equivalent transport as single agent. Conclusions: TFV and DPV were passively transported across Caco-2 cells (non-vectorial and concentration independent). Transport of DRV was vectorial, concentration dependent and affected by transporter inhibitors, suggesting that DRV is a substrate of P-glycoprotein. Tested ARV drugs were unaffected by the presence of double or triple combinations

296 Posters Posters 32: Ethics and the Law P32.0 Ethical Considerations in Implementing a Biometric Co-enrolment Prevention System in Clinical Trials in South Africa P32.02 Fair Subject Selection and HIV Vaccine Efficacy Trials: Canada s Global and Domestic Responsibilities Jayajothi Moodley, Vaneshree Govender, Sarita Naidoo, Dhevium Govender, Gita Ramjee,2, Patrick Charls 3 South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom, 3 South African Medical Research Council, National Information Technology Department, Durban, South Africa Background: Preventing co-enrolment of participants in clinical trials ensures participant safety and data integrity. To facilitate co-enrolment checks, a Biometric Co-enrolment Prevention System (BCEPS) was implemented in 200 by the SAMRC National Information technology (IT) Division, in collaboration with the HIV Prevention Research Unit (HPRU). This is a web based system capturing participant s identification details in real time. Here, we report on the ethical considerations in implementing BCEPS across clinical research sites (CRSs). Methods: Ethical approval for use of this system was obtained from MRC Ethics Committee. All participating research organisations signed a memorandum of agreement, prior to use of BCEPS. Participants who screened at the CRSs had their name, South African identity number and fingerprints captured onto the system. This information was verified and updated at all study visits. If a participant attempted to co-enrol at another site, the system flagged this as a potential co-enrolment. Results: Ethical considerations for implementation included ensuring confidentiality and data security, through password protection and fingerprint access by designated staff. Participant fingerprints are stored as encrypted codes which cannot be copied. A participant information sheet is used to ensure the participant s understanding and right to refuse, or to withdraw from BCEPS at any time. Investigator discretion is used to decide if it is safe for the participant to enrol into the study; or continue with study product use without co-enrolment checks. The period of storage of data within the database is up to 5 years as per South African good clinical practice. Conclusions: BCEPS is a novel approach to prevent co-enrolment in clinical trials. By addressing the ethical aspects and compliance with good clinical practice, we are able to ensure that the system protects participant rights and safety, and ensures data integrity. Rika Moorhouse Ottawa Hospital Research Institute (OHRI), Ottawa, ON, Canada Background: HIV vaccine research is a shared global enterprise, with the potential to yield great benefits for populations at risk. To this end, Canada s contributions to domestic and global efforts are yielding important new knowledge and scaling up capacity for biomedical HIV prevention trials in priority settings. However, there remains a gap in bioethics scholarship on Canada s global and domestic obligations to enroll late phase trials of preventive HIV candidate vaccines. Specifically, there is a dearth of literature on how to operationalize ethical standards for biomedical HIV prevention trials to ensure fair subject selection in Canadian settings. During the STEP Study, community gatekeepers in Canada expressed concerns about the moral defensibility of recruitment in communities of marginalized populations including sex workers, Aboriginal people, and people who use illicit drugs. This perception of moral transgression suggests the need for more detailed and documented moral analysis on fair subject selection specific to Canadian populations, and in anticipation of future preventive HIV vaccine trials (HVTs). Methods: I present key ethical considerations for stakeholders to weigh and balance in order to develop moral accounts of fair selection of HVT participants in Canada, with a focus on the principle of justice. Results: There are at least six key ethical considerations for Canadian stakeholders to consider in the fair selection of subjects for HVTs: duty to include; duty to exclude; respect for communities; risk-benefit profile and net risk; public health research; and global distributive justice. Conclusions: This exploration encourages an ethical analysis of fair subject selection that accounts for both domestic and global responsibilities, and that uses a reflective equilibrium approach. More debate, discussion and commentary among HVT stakeholders in Canada is necessary to ensure that key moral tensions have been described, addressed and, where possible, consensus reached. 296 HIV Research for Prevention 204 HIV R4P

297 Posters 32: Ethics and the Law Thursday, 30 October P32.03 HIV and the Law: The Impact of the Law on HIV Research and the Role of Researchers Michael Ulrich Henry M. Jackson Foundation - Division of AIDS, National Institutes of Health, National Institute of Allergies and Infectious Diseases, Division of AIDS, Rockville, MD, United States There is a growing need to fully explore the connection between the law and HIV, and the negative impact the law can have. Internationally, there is an alarming growth in the popularity of anti-gay laws, which are likely to have a significant impact on the spread of HIV. While the myth that HIV is a disease that plagues homosexuals alone has long been proven false, nevertheless, men having sex with men (MSMs) are a high risk population that suffers from stigma and discrimination that often results in them avoiding the health care system or not receiving the treatment they need. These laws only stand to exacerbate this problem. Furthermore, the stereotype that a man who has contracted HIV is gay has unfortunately not been completely eliminated. As such, these laws may cause heterosexual, as well as homosexual, HIV-positive men to avoid the health care system, fearing criminalization and prejudice. In addition to anti-gay laws, there are HIV criminalization statutes that still not been eliminated, with recent legal cases exhibiting the impact that the legal system can have on public health, and the HIV epidemic specifically. The purpose of this presentation is to analyze and educate how these laws and the enforcement and interpretation of them can impact the HIV community and, in fact, hinder the goal of slowing the spread of the disease. By increasing stigma and misconceptions, while reducing education and understanding, the effect of criminalization laws needs to be fully explored and understood to aid in their eventual elimination. Moreover, there is a growing need to explore the roles of researchers and potential obligations they may have in relation to participants and the legal threats they face

298 Posters Posters 33: Evaluation of Novel Compounds in Cell-Based Systems P33.0 P33.02 HIV- Shows Increased Sensitivity to Griffithsin Derivatives Kabamba Bankoledi Alexandre, Karen W. Buckheit 2, Lauren Haugh-Krumpe 3, Brian Constantine 3, Robert W. Buckheit 2, Barry R. O Keefe 3 Council for Scientific and Industrial Researches, Bioscience, Emerging Health Technology, Pretoria, South Africa, 2 ImQuest BioSciences, Frederick, MD, United States, 3 National Cancer Institute, Molecular Target Laboratory, Frederick, MD, United States Background: The lectin griffithsin (GRFT) is a homodimer isolated from the red alga griffithsia sp. GRFT has demonstrated potent and broad anti-hiv- activity across subtypes and is one of the leading HIV- microbicide candidates. The GRFT Derivatives 2MG, 2MG3, 3MG and 4MG are made of arrays of two, three and four monomeric GRFT units, respectively. Methods: We evaluated HIV- subtype A, B and C against 2MG, 2MG3, 3MG, 4MG and GRFT using the TZM-bl neutralization assay. GRFT derivatives were also tested for their inhibition of the cell-to-cell transmission of HIV-. The 234 and 295 glycans, shown to be important in GRFT binding to HIV-, were introduced in the virus by site directed mutagenesis, and their effects on 2MG, 2MG3, 3MG and 4MG binding studied. GRFT resistant viruses were generated by culturing HIV- under escalating concentrations of the lectin. These resistant viruses were then tested for sensitivity to 2MG, 2MG3, 3MG and 4MG. Results: In general 2MG and 2MG3 were as potent as GRFT against all the viruses tested while 3MG and 4MG were more potent against HIV- subtype A and C. GRFT was also less potent than these two derivatives in the inhibition of cell-to-cell transmission of HIV-. Similar to GRFT, the introduction of the 234 and 295 glycans affected HIV- sensitivity to 2MG and 2MG3; while it did not affect 3MG and 4MG neutralization of the virus. Lastly, GRFT resistant viruses showed sensitive to 3MG and 4MG. Conclusions: The 3MG and 4MG derivatives were more potent than GRFT in inhibiting HIV- infection. Also viruses that showed resistance to GRFT remained sensitive to these compounds. It is possible that 3MG and 4MG binding site on the viral envelope is different from that of GRFT given that the 234 and 295 glycans do affect their neutralization of the virus. The data generated from these studies suggests that linking GRFT into arrays of more than two monomeric units increases its potency against HIV-. Antiviral Activity and Mode of Action of Griffithsin against HSV-2 and HPV: Preliminary Studies of a Potential non-arv Combination Microbicide Keith Levendosky, Olga Mizenina, Kyle Kleinbeck, Larisa Kizima, Aixa Rodríguez, Ninochka Jean-Pierre, Melissa Robbiani, Barry R. O Keefe 2, Thomas Zydowsky, José A. Fernández-Romero Population Council, New York, NY, United States, 2 Molecular Targets Laboratory, Center for Cancer Research, NCI at Frederick, Frederick, MD, United States Background: Griffithsin (GRFT) is a promising HIV microbicide candidate. Nixon et. al. have shown that GRFT blocks herpes simplex 2 (HSV-2) infection in a mouse model, proposing inhibition of cell-to-cell spread as the mode of action (MOA). Using in vitro studies we further investigated the MOA of GRFT against HSV-2 and studied its antiviral activity against human papillomavirus (HPV). We also combined GRFT with zinc acetate (ZA) and/or carrageenan (CG) to render a more potent microbicide. Methods: We used XTT assay to define non-cytotoxic concentrations of GRFT, ZA, CG or their combinations. Assays for anti-hiv, anti-hpv and anti-hsv-2 activities were performed in TZM-bl cells or PBMCs using MAGI and p24 ELISA; in HeLa cells using a luciferase assay; and in Vero cells using plaque forming units (pfu) assay. We performed timeof-addition and temperature dependence experiments to differentiate inhibition of viral adsorption from entry. Surface plasmon resonance (SPR) was used to assess GRFT binding to viral glycoproteins and immunohistochemistry was used to determine the specific glycoprotein involved. Antiviral activities of prototype GRFT/CG (GC) and GRFT/ZA/CG (GZC) gels in a vaginal HSV-2 mouse model were evaluated. Results: GRFT shows modest in vitro antiviral activity against HSV-2 G (IC 50 =5.8µg/ml) and HPV 6, 6, 8, 45 PsVs (IC 50 = µg/ml), compared to potent anti-hiv activity (IC 50 =0.7-.4ng/ml). As with HIV, GRFT blocks the entry but not the adsorption of HSV-2 and HPV to target cells. The combined analyses of SPR and immunohistochemistry for HSV-2 gd, suggest that GRFT binds to HSV-2 gd. GC and GZ had synergistic in vitro antiviral activity against HIV and HPV (CI < ). GC and GZC gels significantly reduced (p< 0.05) HSV-2 vaginal infection in vivo when administered up to 2h before challenge with 0 6 pfu/mouse. Conclusions: GRFT blocks HSV-2 and HPV entry to target cells and combination with CG and/or ZA may result in a potent/broad-spectrum non-arv microbicide. 298 HIV Research for Prevention 204 HIV R4P

299 Posters 33: Evaluation of Novel Compounds in Cell-Based Systems Thursday, 30 October P33.03 P33.04 Humic Acids (HA) Strongly Potentiate Anti-HIV Effects of AZT, Griffithsin, and Cyanovirin Eduard Karamov,2, Galina Kornilaeva, Kabamba Alexandre 3, Barry O Keefe 3, Christiane Moog 4, Ian McGowan 5, Gustavo Doncel 6, Irina Zalenskaya 6, Ali Turgiev 2, Alexander Tatarintsev 2 Ivanovsky Institute of Virology, Moscow, Russian Federation, 2 Immunomica LLC, Moscow, Russian Federation, 3 Center for Cancer Research, National Cancer Institute, Frederick, MD, United States, 4 Strasbourg University, Strasbourg, France, 5 University of Pittsburgh Medical Center and Magee-Women Research Institute, Pittsburgh, PA, United States, 6 Eastern Virginia Medical School, CONRAD, Norfolk, VA, United States Background: The objective of this study was to assess the anti-hiv activity of HA and the synergistic potential of their combinations with NRTI or lectin proteins (LP). Methods: Anti-HIV efficacy of HA was evaluated in PBMC, MDM, DC, Caco-2, and HEC-A cells (using R5 HIV- BaL); syncytium formation (using CEM SS) and TZM-bl assays; and in cell-free model systems with recombinant HIV enzymes. Virus replication was detected by p24 HIV- antigen (intracellular and/or released) ELISA. Cytotoxicity was determined using the MTT assay. Results: HA suppressed HIV- Bal replication in MDM with IC 90 =.5 µg/ ml and IC 50 = 0.4 µg/ml. The activity in PBMC and DC was less pronounced (IC 90 values were 6.0 and 20 µg/ml, respectively). CD4-independent entry of HIV- into endometrial HEC-A cells was suppressed with IC 00 = 00 µg/ml and IC 50 = 0 µg/ml, regardless of the dose of the virus (at 0 3 and 0 4 TCID 50 /ml). HA cytotoxicity in this system was low (CC 50 not attainable; ATP and TEER levels remained unaffected up to 00 µg/ ml). Similar results were obtained with colorectal Caco-2 cells. Selectivity indices in the cell-virus systems studied were in excess of HA had no spermicidal activity up to 3 mg/ml (37 0 C, 30 min). HA augmented the antiviral effects of NRTI (AZT) and LP (griffithsin or cyanovirin). Synergistic effects were determined by concentration-matrix antiviral assays. Combination indices (CIs) showed synergy between HA and either AZT (CI = 0.4) or LP (CIs between ). HA affected at least two phases in the life cycle of HIV: () virus entry (inhibition of syncytium formation) and (2) reverse transcription (experiments with recombinant reverse transcriptase). Conclusions: HA hold significant promise as safe and efficacious drugs for the treatment of HIV infection. The observed synergistic effects may have a utility in HIV prevention strategies (HA may be used as a component of vaginal and/or rectal microbicides). Polyanionic Functionalized Carbosilane Dendrimers as Potential Microbicides to Prevent HIV- Sexual Transmission Enrique Vacas-Córdoba, Francisco J. De la Mata 2, Rafael Gómez 2, Marjorie Pion, Mª Ángeles Muñoz-Fernández Hospital General Universitario Gregorio Marañón, Laboratorio InmunoBiología Molecular, Madrid, Spain, 2 Universidad de Alcalá, Departamento de Química Inorgánica, Alcalá de Henares, Spain Background: Topical microbicides are researched as potential tools in order to stop the HIV sexual spreading in all risk groups. Since the majority of clinical trials in HIV- patients have failed, nanotechnology offers novel suitable approaches to develop new microbicidal compounds, such as dendrimers. Methods: In vitro and in vivo studies were performed to evaluate the safety, biocompatibility, anti-hiv ability and mechanism of two polyanionic carbosilane dendrimers. Moreover, the antiviral activity of carbosilane dendrimer/arv combinations against R5, X4 and dual tropic HIV- isolates was evaluated in human primary cells and TZM.bl cell line using Calcusyn software. Results: Sulphated and naphthylsulfonated functionalized carbosilane dendrimers G3-S6 and G2-NF6 are shown as safety and effective compounds against HIV. They impede laboratory and clinical primary HIV- isolates infection in activated PBMC and inhibit HSV-2 infection in vitro. Dendrimers are able to inhibit viral infection at fusion and thus at the entry step. They impede the binding of viral particles to target cells surface and membrane fusion, through the blockage of gp20/ CD4 interaction. In addition, dendrimers can inhibit cell-to-cell HIV transmission and difficult infectious synapse formation. G3-S6 or G2-NF6 did not produce changes in proinflammatory cytokines profile in treated epithelial cells, in PBMC proliferation, microbiota or sperm survival. Moreover, no irritation, inflammation or vaginal lesions were detected in female mice after dendrimers vaginal administration. As well, G3-S6 and G2-NF6 showed a synergistic activity profile with AZT, efavirenz, maraviroc and tenofovir in the majority of combinations tested against X4 and R5 tropic HIV- in cell lines as well as in primary human cells. Conclusions: Carbosilane dendrimers and their combinations with ARV can be effective antiviral agents, supporting further clinical research on these as potential microbicides in the context of blocking HIV- sexual transmission

300 Posters Posters 33: Evaluation of Novel Compounds in Cell-Based Systems P33.05 P33.06 Antiviral Action of Sulfonate Anionic Carbosilane Dendrimer as a Topical Microbicide against HIV Infection Daniel Sepúlveda-Crepo, Javier Sánchez-Rodriguez, María Jesús Serramia, Ana López, Esther Alonso, Rafael Gomez 2, Francisco Javier De La Mata 2, Jos Luis Jiménez, Mª Ángeles Muñoz- Fernández Hospital General Universitario Gregorio Marañón, IISGM, Networking Research Center of Bioengineering, Madrid, Spain, 2 Universidad de Alcalá, Campus Universitario, Alcalá de Henares, Networking Research Center of Bioengineering, Biomaterials and Nanomedicine (CIBER- BBN), Madrid, Spain Background: Microbicides include moderately specific macromolecular anionic polymers that block HIV and other STIs, and antiretrovirals (ARVs) that inhibit HIV entry and reverse transcription. Based on nanotechnology, we show a novel water-soluble anionic carbosilane dendrimer, 2G-S6, as an advantageous molecule against HIV-infection. Methods: 2G-S6 was synthesized containing sulfonate peripheral groups. Cellular in vitro or in vivo models were used to study the safety, biocompatibility and anti-hiv ability of 2G-S6. Study of dendrimer/ ARVs effects and the EC 50 were performed using Calcusyn software. Results: 2G-S6 shows as safety and effective compound against HIV- and HIV-2 with great potential as topical microbicides. 2G-S6 has a great capacity to block HIV entry inside epithelia cells derived from uterus and vagina due to 2G-S6 protect the epithelial monolayer from cell disruption. Also impede laboratory and clinical primary X4, R5 and X4/R5 HIV isolates infection in activated PBMCs. 2G-S6 does not change the proinflammatory cytokines profile in treated epithelial cells, in PBMC proliferation, on microbiota or sperm survival. We research the mechanism of action and shown that 2G-S6 inhibits HIV infection at fusion and thus and entry step. 2G-S6 impedes the binding of HIV particles to target cell surface and membrane fusion, through the blockage of gp20/cd4 interaction. Interestingly, 2G-S6 in combination with tenofovir or maraviroc obtains 00% HIV-inhibition and displayed a synergistic profile at low micromolar doses against a broad-spectrum of HIV strains in TZM.bl. Also, no irritation or vaginal lesions were detected in female rabbit genital tracts and in CD (ICR) mice after daily different concentrations of 2G-S6 for 7 days. The proof of concept is been performed by using 2% (W/V) hydroxylethyl cellulose gel (HEC) with 3% of 2G-S6 in humanize mice. Conclusions: 2G-S6 is effective to inhibit HIV infection and transmission within genital mucosa. Identification of a Novel Acylguanidine-based Inhibitor of HIV- Replication Philip M. Mwimanzi, Ian Tietjen 2, Aniqa Shahid, Scott C. Miller 2, David Fedida 2, Zabrin Brummer,3, Mark Brockman,3 Simon Fraser University, Faculty of Health Sciences, Burnaby, BC, Canada, 2 University of British Columbia, Vancouver, BC, Canada, 3 British Columbia Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada Background: Increased access to therapy has reduced HIV- morbidity and mortality, but new drug classes would further enhance treatment options and counter resistance. Acylguanidine-based molecules are active against diverse viruses. One member of this group, BIT225, is reported to inhibit HIV- by blocking the viroporin function of Vpu, however its clinical utility is limited by toxicity. We investigated the anti- HIV- activity of a novel acylguanidine compound, SM. Methods: We used a GFP-reporter T cell assay to test SM s ability to inhibit replication of NL4-3 and four recombinant strains encoding major NRTI- and/or NNRTI-resistance mutations in Pol (e.g. D67N and/or K03N, respectively). Viruses were cultured in the presence of SM (0-00µM), AZT (NRTI; 00nM) or EFV (NNRTI; 00nM) and infected GFP+ cells were monitored by flow cytometry. Drug activity was assessed on day 6 compared to media controls. Cytotoxicity was evaluated using ViaCount (Millipore). NL4-3 was also passaged in 00µM SM; three independent drug-resistant strains were isolated and sequenced. Results: SM inhibited NL4-3 in a dose-dependent manner between 0µM and 00µM. HIV-infected cells were reduced >98% at 00µM (44.3% [ ] in absence vs % [ ] in presence of drug). Similar activity was observed against NRTI and NNRTI resistance strains (>95% reduction in all cases). In contrast to BIT225, SM was not toxic at any dose tested. Notably, SM-resistant strains encoded mutations in the transmembrane of Vpu, including a 5AA deletion, a substitution or insertion of a stop codon at highly conserved W22. Conclusions: SM is a novel compound that can inhibit wild type as well as NRTI- and NNRTI-resistant HIV- strains, indicating that it has a different mechanism of action than current drugs. Resistance patterns suggest that SM s target is Vpu, but additional studies are necessary to explore the mechanism of this promising prototype. Funded by CIHR and the Michael Smith Foundation for Health Research 300 HIV Research for Prevention 204 HIV R4P

301 Posters 33: Evaluation of Novel Compounds in Cell-Based Systems Thursday, 30 October P33.07 Microbicide, SsALF-24 Prevents HIV Infection through the Blockade of gp20 Binding to CD4 Receptor Kvr Reddy, Ankit Shroff National Institute for Research in Reproductive Health, Molecular Immunology & Microbiology, Mumbai, India P33.08 Design and Discovery of Pyrazole and Pyrimidine as Novel Class of Potent Nonnucleoside Reverse Transcriptase Inhibitors (NNRTIs) Udaya Pratap Singh, Hans Raj Bhat Sam Higginbottom Institute of Agriculture, Technology & Sciences, Deemed University, Drug Design & Discovery Laboratory, Department of Pharmaceutical Sciences, Allahabad, India Background: Thirty years after its first case, HIV/AIDS has become a pandemic and is raging in many parts of the world. UNAIDS report of 20 states that there were.7 million deaths through AIDS and 2.5 million were newly infected by the virus. Inspiring responses by researchers and doctors have resulted in survival of marginal populations of patients on potent antiretroviral therapy. Thus, preventing entry of the virus into host is the best way to tackle this infection. A major route of infection is the sexual route. Hence, developing a safe vaginal product that can block HIV entry into the host is of utmost importance. Methods: Anti-HIV activity of SsALF-24 was analyzed by p24 ELISA, cell-cell fusion and Luciferase assays. Binding efficiency of SsALF-24 to gp20 was determined using Surface Plasmon Resonance (SPR) spectroscopy. Toxicity of SsALF-24 on vaginal epithelial cells was assayed by Trans-Epithelial Electrical Resistance (TEER). Cytokine profile of human vaginal epithelial cells (VK2/E6E7) on interaction with SsALF-24 was determined by estimating the levels of Interleukin-6 (IL- 6), Interleukin-8 (IL-8), Monocyte Chemotactic Protein - (MCP-) and Interkeukin-α (IL- α) using ELISA, Western blot, Flow cytometry, q-pcr. Results: SsALF-24 is derived from SsALF (Scyalla serrata Anti- Lipopolysaccharide factor). p24 ELISA, cell-cell fusion and Luciferase assays show that SsALF-24 binds to gp20 and thereby prevents the binding of the latter to CD4 receptor. This blocks the downstream events of infection. SsALF-24 is not toxic to vaginal epithelial cells as demonstrated by the results of TEER and microsphere experiments. Cytokine profile of vaginal epithelial cells on exposure to SsALF-24 indicates that it may have anti-inflammatory activity. Conclusions: SsALF-24 binds of gp20 and prevents the latter s binding to CD4 receptor and subsequent viral entry. Hence, SsALF-24 may be developed as a microbicide that prevents viral entry. Background: Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs) considered as structurally diverse group of compounds, act by inhibiting the reverse-transcriptase (RT) enzyme in an allosteric mode by binding to the polymerase active site causing a distortion of the catalytic aspartate triad in a non-competitive fashion. Consequently, a wide chemical opportunity exists, due to the flexibility of the NNRTI binding pocket (NNIBP) in the RT. In continuation of our ongoing efforts in the discovery of economic NNRTI agents, herein, we wish to report a novel class of pyrazoles and pyrimidines in order to obtain more effective candidates. Methods: A novel series of pyrazole and pyrimidine compounds has been developed via cyclocondensation reaction. These molecules have been subsequently tested for anti-hiv activity using TZM-bl cell lines along with Luciferase expression profile of the TZM-bl cells after infecting with NL4.3 virus and MTT assay for the cytotoxicity determination. Results: In Anti-HIV assay, molecules having pyrazole amine with distant position of electron withdrawing group showed 9-98 % inhibition. Further these compounds in Luciferase assay showed considerable inhibition of infection. While in cytotoxicity assay, it was observed that an increase in the concentration of most active compounds from 25 mg/ml to 25 mg/ml did not appreciably lower the percentage of cell viability. A close inspection of the best docked pose of most active compound clearly establish that it attained a horseshoe-like conformation and interaction with the Tyr8 and Tyr88 of the p66 subunit in the NNIBP. Conclusions: As a concluding remark, we have developed a novel series of compounds with potent anti-hiv activity. It was confirmed that the designed molecules have the possibility of introducing chemical diversity around the core skeleton to generate new, potent molecules. 30

302 Posters Posters 33: Evaluation of Novel Compounds in Cell-Based Systems P33.09 Developing an Effective Rectal Microbicide: Inhibiting HIV Transmission in Human Colorectal Tissue and Humanized Mice with CD4 Aptamer-siRNA Chimeras Lee Adam Wheeler, Judy Lieberman Harvard Medical School, Boston, MA, United States P33.0 Glycolysis Inhibitors as Potential Anti-HIV Compounds Elijah Songok, Benjamin Nzau, Mark Wainberg 2, Frank Plummer 3, Solomon Mpoke Kenya Medical Research Institute, Centre for Virus Research, Nairobi, Kenya, 2 McGill University, Montreal, QC, Canada, 3 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada Background: The continued spread of the HIV epidemic underscores the need to interrupt transmission. One attractive strategy is a topical microbicide. We previously showed that intravaginal application of sirnas targeted to CD4+ cells protected humanized mice from sexual transmission, but to date no microbicide candidates have proven effective for the prevention of the rectal transmission of HIV. Methods: Here, we use chimeric RNAs composed of a CD4-specific aptamer fused to sirnas targeting the HIV coreceptor CCR5, gag, and vif to knockdown target gene expression in CD4+ cells in human colorectal tissue and in vivo, by employing a small animal model of HIV transmission. Results: Here, we demonstrate that CD4 aptamer-sirna chimeras (CD4-AsiCs) maintain stability for over 24h in a colorectal lavage, and over 36h when suspended in an FDA-approved hydroexthylcellulose (HEC) gel formulation. We provide evidence that CD4-AsiCs effectively deliver sirnas to CD4+ cells in human colorectal tissue explants, and in the colorectal tissue of humanized mice without stimulation of an inflammatory response after administration. When applied intra-rectally to humanized mice, CD4-AsiCs in solution provide substantial but incomplete protection from HIV transmission. When administered in an HEC gel formulation however, all treated mice are protected against HIV transmission for up to 8 weeks. Conclusions: From these data, we conclude that topical CD4-AsiCs administered in a HEC gel formulation could be used as the active ingredient of a microbicide to prevent the rectal transmission of HIV. Background: In a bid to determine correlates of HIV protection, a whole blood microarray study revealed that HIV exposed seronegative (HESN ) sex workers have a significantly lowered glycolyis rate. This suggested that reduction of cell glycolytic activity may be protective or beneficial against HIV Infection. This phenomenon has been noted among cancer cells, where compounds that lower glycolysis rate have been developed as potential therapeutics against malignant carcinoma. Our observation among HESN suggests that similar to cancer cells, HIV maybe utilizing the glycolytic pathway for its energy needs. We have carried a preliminary in vitro analysis to determine if the glycolytic inhibitor,s lonidamine LND and 2 Deoxyglucose (2DG), has an anti-hiv effect. and a potential therapeutic against HIV. Methods: Viral Reverse Transcriptase (RT) was measured from culture supernatant using RT assay on pretreatment (4hr drug exposed followed by 2hr virus infection) and co treatment. Cell cytotoxity on mock infected C866 cells was done in parallel with RT assay using Trypan blue exclusion test and Vi-cell (Beckmancounter) cell viability analyser. Results obtained were normalized and presented as percentages. Results: Lonidamine co treatment at 00 µm resulted in more than 76% HIV inhibition while 2-DG concentrations at 5mM had HIV inhibition percentage above 90% inhibition. Cell toxicity assessment at similar concentrations resulted in 75% reduction in actual cell count with LND co treatment. The 2-DG component maintained a cell proliferation of more than 90%. Conclusions: Glycolytic inhibitors 2-DG and LND have potential anti-hiv activity. However it suggests a better safety profile of 2-DG at effective inhibitory concentrations. 302 HIV Research for Prevention 204 HIV R4P

303 Posters 33: Evaluation of Novel Compounds in Cell-Based Systems P33. LB Targeting the Glucocorticoid Receptor with Selective Modulators for Prevention against HIV- Infection Thursday, 30 October Chanel Avenant, Michele Tomasicchio, Roslyn Michelle Ray, Andrea Du Toit, Yashini Govender, Hazel Hunt 2, Janet Patricia Hapgood University of Cape Town, Cape Town, South Africa, 2 Corcept Therapeutics, Storrington, United Kingdom Background: Due to their anti-inflammatory properties, glucocorticoids (GCs) are widely used therapeutically. In the female genital tract, inflammation correlates with increased HIV infection, suggesting the use of GCs in multipurpose prevention therapy. However, the effects of different GCs and the role of the glucocorticoid receptor (GR) in HIV infection and the mechanisms involved are unknown, and are likely to be relevant to HIV- prevention strategies. Methods: PBMCs or End/E6E7 endocervical epithelial cells were incubated with different GCs. PBMCs were infected with virus prepared from HIV- BAL-LUC IMCs. Inflammatory markers were measured by qrt- PCR or FACs. GR function was assessed by western blotting, ChIP and sirna experiments. Results: Infection assays in PBMCs showed that while Dex and CORT376 increased infection, pre-treatment with the GR antagonist RU486 and a selective GR modulator CORT08297 did not increase, but potentially had a protective effect, consistent with a role for the GR in HIV- infection. Differential regulation of GILZ, IL6 and RANTES genes by different GCs indicated a mechanism whereby some GCs increase, while others protect against HIV infection. Furthermore, Vpr was found to activate the GR, resulting in repression of both basal and induced cytokine genes in the absence of GCs. Conclusions: HIV exploits the host GR to favor viral infection via several strategies. In the absence of GCs, the viral protein Vpr modulates GR activity to change expression of basal and induced cytokine genes, suggesting that GR levels play a critical role in HIV pathogenesis. GR agonists repress immune function, while at the same time increase HIV- infection, suggesting their use should be avoided in high risk areas. Selective GR modulators exhibit variable effects on HIV- replication and gene-specific effects on expression of cytokine genes. Some show potential application for combination therapy in the female genital tract by both repression of RANTES and repression of HIV- infection

304 Posters Posters 34: Glycans and Antibody Effector Functions P34.0 Identification of Specificities of Broadly Neutralizing Plasma Antibodies Obtained from HIV- Clade C Infected Indian Donors Jayanta Bhattacharya, Shilpa Patil, Sweety Samal, Saikat Boliar, Tripti Srivastava, Manish Bansal, C Ritcher King 2, K G. Murugavel 3, Suniti Solomon 3, Bimal K. Chakrabarti HIV Vaccine Translational Research Laboratory, THSTI-IAVI HIV Vaccine Design Program, Translational Health Science & Technology Institute, Gurgaon, India, 2 International AIDS Vaccine Initiative, New York, NY, United States, 3 YRG Care, Chennai, India P34.02 HIV- gp20 Impairs B Cell Proliferation by Inducing TGF-β Production and FcRL4 Expression Claudia Cicala, Katija Jelicic, Raffaello Cimbro, Fatima Nawaz, Xin Zheng 2, Jun Yang 2, Richard Lempicki 2, Donald Van Ryk, Jocelyn Ray, Joseph Hiatt, Catherine Schwing, Danlan Wei, Massimiliano Pascuccio, John Kehrl, James Arthos, Anthony S. Fauci NIH/NIAID, LIR, Bethesda, MD, United States, 2 NIH/NIAID, Laboratory of Immunopathogenesis and Bioinformatics, SAIC, Frederick, MD, United States Background: A number of broad and potent neutralizing human monoclonal antibodies (mabs) against diverse regions of HIV- Env have been reported. However, it is not known whether HIV- clade C infections in India mount a neutralizing antibody response against any of those known epitopes. In the present study, we examined () plasma samples obtained from anti-retroviral (ART) naïve chronically infected Indian donors for their capacity to neutralize a diverse virus panel representing different subtypes and (2) we mapped the specificity of the epitopes targeted. Methods: Two hundred plasma samples were assessed for their capacity to neutralize Env-pseudotyped viruses in a TZM-bl cell assay. CD4 binding site (CD4bs), N60, N332-glycan and MPER directed neutralizing activity of the analyzed plasma samples were determined by using TriMut core protein, N60A/N332A mutant and HIV2/HIV- chimeric viruses respectively. Results: 0/200 (5.5%) plasma antibodies were found to neutralize >60% of the 50 panel viruses of distinct geographical origin and belonging to clades A, B. C, B/C,A/E and A/G with a median ID50>200. Amongst these ten broadly neutralizing plasma (BNP) samples, two (%) were found to be highly potent (median ID50 > 400). Interestingly, none of the 200 plasma samples showed any specificity to CD4 binding site (CD4bs), indicating a possible inability of clade C strains circulating in India to elicit CD4bs directed antibodies. In addition, none of the neutralizing activity by the BNP samples was dependent on N60 and N332 glycans. Three of the BNP samples showed specificity to the gp4 membrane proximal external region (MPER). This MPER reactivity was different than that of the known MPER neutralizing mabs. Conclusions: Our data indicates that the BNP samples obtained from Indian donors likely target novel epitopes and would pave way towards identification of new vulnerable site/s on HIV- envelope. Background: During the early stages of HIV infection the immune system of the infected individual is impaired. Among the defects described is the impairment of normal B cell function, that includes a significant delay in the development of the anti-hiv humoral immune response. The mechanisms underlying this delay are not fully understood. In the present study, we asked whether gp20-induced signaling through α 4 β 7 on B cells could disrupt their function. We show that gp20 binds and signals through α 4 β 7 on naïve B cells, resulting in an abortive proliferative response. Methods: We performed microarray analysis, RT-PCR, CFSE proliferation assay, ELISA, FACS analysis, cell cycle analysis on freshly isolated primary human B cells. B cells were stimulated with CpG, anti-human IgM and anti-cd40 mab, in presence or not of recombinant HIV- gp20. Results: gp20 directly impaired the activation and proliferation of naive B cells by releasing TGF-β, which in turn up-regulated expression of the inhibitory IgA receptor FcRL4. Co-culture of B cells with HIV-infected autologous CD4 + T cells also increased B cell FcRL4 expression. Treatment of stimulated B cells with gp20 decreased the expression of the co-stimulatory receptor CD80 but not CD86. gp20 impaired proliferation of naïve B cells stimulated with both a TI and a TD stimulation. gp20 affected the responses of memory B cells to TI and TD stimulation in a relatively modest way compared with the suppressive effect we observed for naïve B cells. Conclusions: These findings indicate that, in addition to mediating chronic immune activation, viral proteins can contribute directly to HIV-associated B cell dysfunction. These studies have implications for understanding the immune-pathogenic mechanisms of HIV- infection, particularly the ability of high levels of viremia, observed during acute HIV infection, to blunt the early antibody response to HIV. 304 HIV Research for Prevention 204 HIV R4P

305 Posters 34: Glycans and Antibody Effector Functions Thursday, 30 October P34.03 Shielding of the HIV- Envelope Membrane Proximal External Region (MPER) from Antibody P34.04 The Dynamics of HIV- gp20 N-linked Glycans in the Context of Broadly Crossneutralizing Antibodies Rajesh Abraham Jacob,2, Thandeka Moyo,2, Fatima Abrahams, Berta Grau Pujol, Jeffrey R. Dorfman,2 International Centre for Genetic Engineering and Biotechnology, Cape Town, South Africa, 2 University of Cape Town, Division of Immunology, Cape Town, South Africa Background: The Membrane Proximal External Region (MPER) of HIV- gp4 envelope is an attractive vaccine target. We have previously identified 253- (CRF_02AG) as unusually neutralization-resistant and studied its sensitivity to anti-mper antibodies. Methods: Antibodies reactive against the 253- MPER were identified using a chimeric HIV-2 virus displaying the 253- MPER sequence in place of its own. The presence of dominant anti-mper neutralizing antibodies was tested by assessing the reduction in neutralization after depleting MPER-specific antibodies. We also studied MPER swap viruses between 253- and , a subtype-matched virus that is sensitive to anti-mper antibodies in cohort sera. Results: We found that 253- is rarely neutralized by anti-mper antibodies in sera from individuals HIV-infected >yr. However, recognition of that MPER in the HIV-2 chimera was common: 9/77 (9%) sera recognized 253- s MPER in the chimeric virus but not the original 253- virus. At least 3/9 of these sera neutralized other HIV- isolates via MPER, indicating that these anti-mper antibodies were not generally defective for neutralization of HIV- and that many viruses do not share this pattern of resistance to anti-mper antibodies. Importantly, sensitivity of /253- MPER swap viruses to these 9 sera and to MPER-specific mab Z3e is primarily controlled by sequences outside the MPER region. Several epitopes were recognized by the sera, suggesting that conformational differences of MPER between the native virus and the HIV-2 chimera are unlikely to explain 253- s neutralization resistance. Conclusions: We consider several explanations for our data and propose that the most parsimonious explanation is obstruction of access to MPER prevents anti-mper neutralization of In this case, only the limited proportion of anti-mper antibodies that can penetrate similar obstructions would be able to provide the protection against the large number of HIV- variants that would be desirable in a vaccine. Ereshia Gabier, Natasha Wood, Simon Travers University of the Western Cape, SANBI, Cape Town, South Africa Background: HIV- prevention through vaccination remains challenging due to the high mutation rate and variability amongst viral strains meaning that the identification of epitopes that elicit a truly broadly cross clade neutralizing virus is an immensely complex task. Identifying vaccine targets is further complicated by the presence of N-linked glycans on the surface of gp20 that protects the virion from recognition by the host immune system. Recently, however, broadly cross-neutralizing (BCN) antibodies have been identified that recognize specific glycans on the surface of gp20 and result in the neutralization of a broad panel of HIV viruses. Studies have identified that the glycans bound at either N332 or N334 in the C3 region at the base of the V3 loop are critical for susceptibility to, or evasion of neutralisation. The structural mechanism of this critical process is not yet fully understood. Methods: Using the full gp20 structure, containing both the V/ V2 loops and V3 loop, we performed a molecular dynamics study to model the effect of the movement of glycosylation from 332 to 334 on both the glycan shield and the underlying protein and, thus, on the susceptibility to neutralization. Results: We have identified features of the glycan-glycan, as well as glycan-protein, interaction that may appear to play a role in the recognition of these epitopes by BCN antibodies as well as in the evasion of neutralization. Conclusions: Our current research therefore contributes to the understanding of the role of N-linked glycosylation in the context of BCN antibodies and their associated epitopes

306 Posters Posters 34: Glycans and Antibody Effector Functions P34.05 Post-attachment Neutralization by Singlechain Variable Fragment (scfv) from an Anti-V3 Monoclonal Antibody with Crossreactivity Yasuhiro Maruta, Takeo Kuwata, Kazuki Tanaka, Yusuke Nakahara 2, Kristel Ramirez, Muntasir Alam, Yoshika Egami, Izumi Hirata, Yoshiaki Suwa 2, Hiroshi Morioka 2, Shuzo Matsushita Matsushita Project Laboratory, Center for AIDS Research, Kumamoto University, Kumamoto, Japan, 2 Department of Analytical and Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan P34.06 HIV- Neutralisation Breadth Is Positively Associated with Presence of Anti-MPER Antibodies and Not of Anti-PG9/6-site Antibodies Thandeka Moyo,2, Rajesh A. Jacob,2, Michael Schomaker 3, Berta Grau 4, Fatima Abrahams 4, Jeffrey R. Dorfman,4 Division of Immunology, University of Cape Town, Cape Town, South Africa, 2 International Centre for Genetic Engineering and Biotechnology, Cape Town, South Africa, 3 Centre for Infectious Disease Epidemiology & Research, Cape Town, South Africa, 4 International Centre for Genetic Enginnering and Biotechnology, Cape Town, South Africa Background: The neutralization antibody response against HIV- is crucial for controlling HIV- infection. Such is the case of the epitope in the V3 loop that is exposed after binding of gp20 to CD4. However, it is difficult for anti-v3 IgGs to bind their epitope following CD4-gp20 interaction because IgG is too large to access the close physical proximity of gp20 and the cellular membrane. Methods: We constructed scfv from C0, an anti-v3 monoclonal antibody with cross-reactivity. The C0 scfv was produced in E.coli, purified and refolded by on column refolding process. In addition to the usual single cycle neutralization assay, we employed temperature regulated neutralization assay (TRN assay) and neutralization assay using SOS JR-FL virus (SOS assay) to evaluate post-attachment neutralization using TZM-bl cells as a target. TRN assay and SOS assay allows determining whether scfv can access efficiently to the narrow space between HIV- and the V3 epitope of the envelope after attachment of the virus to CD4. Results: Neutralization activity of C0 scfv was greater than that of IgG counterpart against JR-FL, YU2 and 89.6 strains of HIV- (IC 50 values of scfv were about 5, 3 and 3 times better than values with IgG, respectively). In addition, C0 scfv neutralized viruses resistant for neutralization by C0 IgG, such as SVPB8 (IC 50 of IgG; >50 µg/ml, scfv; 5.23 µg/ml). TRN assay results showed that neutralization capacity of IgG was considerably reduced after the virus bound to CD4. On the other hand, neutralization activity of scfv at TRN assay was equivalent to that at usual single cycle neutralization assay. In SOS assay, C0 scfv neutralized SOS JR-FL, but C0 IgG did not. Conclusions: These results suggest that the anti-v3 scfv can neutralize HIV- even after attachment of the virus to the target cells. For this reason, the use of scfv may be one of the promising strategies to overcome neutralization resistance of HIV-. Background: The Membrane Proximal External Region (MPER) and the PG9/6-site are both potential targets for anti-hiv- antibody-based vaccines. The MPER of the gp4 subunit of the envelope glycoprotein is relatively conserved and plays a key role in viral fusion with target cell membranes. The PG9/6-site is a quaternary epitope which is expressed on trimeric envelope protein spanning conserved regions of variable loops of the gp20 subunit. The PGT/2G2 antibodies recognise an epitope made primarily from a glycosylation at sites 30 and/or 332. Methods: We explored associations between neutralisation breadth and presence of antibodies directed against these sites in sera from 77 antiretroviral-naïve HIV--infected (>yr) individuals. Anti-MPER antibodies were detected using chimeric 732A HIV-2 viruses engrafted with three different HIV- MPER sequences: a consensus subtype C sequence or the MPER sequence from Yu2 (B) or 253- (CRF02_ AG) viruses. Dominant anti-pg9/6-site antibodies were detected by determining what proportion of neutralisation of any of 3 viruses was abrogated by mutations at positions 60 or 69 within the PG9/6 site. Results: Anti-MPER activity and activity directed against epitopes overlapping the PG9/PG6 site were common. Neutralisation breadth of the MPER-recognising sera was significantly higher than that of the non- MPER recognising samples. We found no evidence for an equivalent association for sera containing anti-pg9/6 site antibodies. Variability within the MPER, measured by analysing 3829 envelope sequences, was substantially lower than that in the PG9/6-site and other broadly neutralising antibody targets. A similar comparison for the PGT/2G2 specificity will also be presented. Conclusions: Our data suggest that inducing anti-mper antibodies by vaccination is more likely to be productive than inducing broadly neutralising anti-pg9/6 site antibodies, even if PG9/6-site immunogen models can be engineered. 306 HIV Research for Prevention 204 HIV R4P

307 Posters 34: Glycans and Antibody Effector Functions Thursday, 30 October P34.07 P34.08 Identification of Key Determinants for the Unusual Neutralization Sensitivity of the MW Env Zakiya Qualls, James Theis, James Robinson 2, Abraham Pinter Rutgers Biomedical and Health Sciences, Public Health Research Institute, Newark, NJ, United States, 2 Tulane University, Health Sciences Center, New Orleans, LA, United States Background: The V3 domain of gp20 is the most immunogenic region of the functional Envelope (Env) spikes found on the surface of a HIV virion. However, in most cases high affinity antibodies (Abs) developed to this region are unable to neutralize the majority of circulating isolates as the V3 loop is usually effectively masked. ConC, the virus encoded by the clade C consensus sequence, is extremely resistant to neutralization by V3 Abs. On the other hand, the clade C tier a virus MW965, is the most neutralization sensitive viral isolate described to date and can be neutralized by a wide range of these Abs. Compared to ConC, MW965 has a longer more highly glycosylated V/V2 region, which usually correlates with greater masking. It is thus unclear why this isolate is so sensitive. Methods: To identify key neutralization determinants in the MW965 Env, chimeric Env constructs were produced between this Env and ConC using various cloning techniques. Neutralization sensitivity to several α-v3 and α-quaternary neutralizing epitope (QNE) Abs was determined using pseudovirus produced from Env expressing plasmids and pnl4-3. luc.r-e-. Reversal of neutralization phenotype indicated the identification of a determinant(s). Results: The construction of a chimeric Env using the SF62 (unmasked) backbone and MW965 V/V2 revealed that this V/V2 is in fact capable of masking the V3. This suggests that another Env region(s) is responsible for this neutralization sensitivity despite the masking properties of the V/V2 domain. Neutralization assays using additional ConC and MW965 chimeric Env constructs, ruled out the C-V3 and gp4 Env regions and narrowed several key determinants to the C3 and C5 Env regions. Conclusions: The identification of specific residues in these domains defines a novel mechanism for regulating neutralization sensitivity in a circulating isolate. Additionally, several of the chimeric Env constructs may provide additional insights for designing immunogens capable of eliciting effective antibody responses. Strain Specific Anti-HIV Antibody Evolution during Acute Infection and Viral Escape Cathrine Scheepers,2, Dshanta Naicker, Chaim Schramm 3, Zhizhang Sheng 3, Arshad Ismail, Salim S. Abdool Karim 4, Bronwen Lambson, Penny Moore,2,4, Lawrence Shapiro 3, Lynn Morris,2,4 Center for HIV and STIs, National Institute for Communicable Diseases (NICD), Sandringham, South Africa, 2 University of the Witwatersrand, Virology, Sandringham, South Africa, 3 Columbia University, Biochemistry and Molecular Biophysics and Department of Systems Biology, New York, NY, United States, 4 Center for the AIDS Programme of Research in South Africa (CAPRISA), Durban, South Africa Background: We previously identified an individual (CAP88), from the CAPRISA 002 cohort, with potent strain-specific neutralization. This response mapped to the C3 region of gp20 and was detected at weeks post-infection (wpi) and then later waned co-incident with viral escape. A monoclonal antibody (CAP88-CH06) was isolated at 34 wpi which utilized the IGHV4-39*0, D3-3*0 and J4*02 genes, had 5.9% divergence from germline, and CDRH3 length of 7 amino acids. This study examined the evolution of the CAP88-CH06 heavy chain immunoglobulin genes over 2 weeks, starting from weeks of infection. Methods: RNA and DNA were extracted from PBMCs from donor CAP88 at four time-points (, 7, 38 and 2 wpi). The heavy chain VDJ regions of IGHV4-39 gene were PCR amplified and sequenced by Illumina MiSeq. The resulting sequences were blasted against a database of germline IGHV and IGHJ sequences from IMGT and compared to the CAP88-CH06 sequence. Results: We detected ~3,500 sequences that were highly related to CAP88-CH06. The majority (75%) of these were from RNA at wpi (,05 sequences, 39%), 7 wpi (,625, 6%) and sequence each in 38 wpi and 2 wpi. This corresponded to the antibody response which first appeared at wpi, peaked at 26 wpi and by 54 weeks had declined. Most of the DNA sequences were from wpi (n=76) followed by 38 wpi (n=3) and fewer than 0 from 7 wpi and 2 wpi. The 38 wpi DNA sequences were closely related to the wpi sequences of both RNA and DNA. Conclusions: We have identified clonally related antibody sequences from 4 different time-points from CAP88 in both RNA and DNA. The frequency of sequences in RNA corresponded with plasma neutralizing antibody titres. DNA-derived sequences from later time-points clustered in phylogenetic trees with RNA-derived sequences from earlier timepoints, suggesting that they were from the memory B cell compartment

308 Posters Posters 34: Glycans and Antibody Effector Functions P34.09 P34.0 Engineering Antibodies to Enhance Activity and Increase Half-life Stuart A. Sievers, Sonal N. Patel, Kathleen Bennett, Florian Klein 2, Michel C. Nussenzweig 2, Pamela J. Bjorkman California Institute of Technology, Biology and Biological Engineering, Pasadena, CA, United States, 2 Rockefeller University, Laboratory of Molecular Immunology, New York, NY, United States Background: HIV/AIDS remains one of the most serious current threats to global public health. Although anti-hiv drugs have been effective among the wealthiest populations, a vaccine and/or new methods to prevent infections are needed to control HIV globally. Strategies to combat HIV- require structural knowledge of how antibodies recognize HIV envelope proteins and how the immune system eliminates viruses. Until recently, only a small number of broadly neutralizing antibodies against HIV- had been characterized, and the immunological basis for their breadth and potency remains poorly understood. However, it was recently demonstrated that antibodies could be engineered to greatly enhance their breadth and potency (Diskin et al., Science 20). Unfortunately, this and other engineering efforts have resulted in a decrease in antibody half-life in mouse and non-human primate models. This decrease in half-life correlates with an increase in reactivity to a variety of antigens, termed polyreactivity. Methods: In order to make better targets for passive delivery therapies, we are working to increase the half-life of antibodies while maintaining their breadth and potency using a variety of computational and structure-based techniques. One technique involves reducing the spatial aggregation propensity, in which an algorithm finds dynamically exposed hydrophobic patches on the surface of proteins (Chennamsetty et al., PNAS 2009). To this end, we have constructed several mutations in regions that have been predicted to have high aggregation propensities, and have tested them for polyreactivity and potency in neutralization assays. Results: Initial results show that these novel reagents have reduced polyreactivity, yet they still maintain their potency in in vitro neutralization assays. Conclusions: We are currently pursuing in vivo experiments in mice to further understand the relationship between antibody potency, polyreactivity, and half-life. Antibody-based PrEP and Cross-reactivity Kevin J. Whaley, Steve Hume 2, Larry Zeitlin 3 Mapp Biopharmaceutical, San Diego, CA, United States, 2 Kentucky Bioprocessing, Inc., Owensboro, KY, United States, 3 Mapp Biopharmaceutical, Inc., San Diego, CA, United States Background: Broadly neutralizing antibodies (bnabs) are being developed for topical and systemic pre-exposure prophylaxis. Since some bnabs (e.g. 4E0) have been reported to interact with non-viral epitopes, the cross-reactivity of bnabs is an important safety parameter to be documented in regulatory submissions. The objective of this study was to determine the cross reactivity of Nicotiana (-N) manufactured anti-hiv bnabs 4E0-N and VRC0-N, and anti-hsv glycoprotein D bnab HSV8-N with cryosections of human tissues. Methods: In order to detect binding, the antibodies were biotinylated and applied to cryosections of normal human tissues (3 donors per tissue) at two concentrations (2-20 µg/ml). Commercially available Synagis was biotinylated and used as a control. The study was GLP compliant. Results: 4E0-N variably stained a variety of tissue elements in the human tissue panel. VRC0-N also produced staining of tissue elements; however, the staining with VRC0-N was generally present in fewer tissues and with reduced intensity and frequency. No staining with HSV8-N or Synagis was observed in the human tissue panel examined. Conclusions: The majority of observed staining was cytoplasmic in nature, which is of little toxicologic concern since the cytoplasmic compartment is generally considered to be inaccessible to antibodies administered in vivo. The toxicologic concern for the observed staining of extracellular elements in selected tissues with 4E0-N and VRC0-N is unknown. Since no staining was observed with HSV8-N the binding of VRC0-N cannot be attributed to the Nictotiana-based manufacturing system. 308 HIV Research for Prevention 204 HIV R4P

309 Posters 34: Glycans and Antibody Effector Functions Thursday, 30 October P34. LB Superinfected Patient Pseudovirus Exhibits Resistance to Broadly Neutralizing Antibodies, but Sensitivity to Autologous Plasma Postsuperinfection Ralf Duerr, Sonal Soni, Colleen Courtney, Josephine Meli 2, Johnson Ngai 2, Luzia Mayr, Phillipe Nyambi,3 New York University - School of Medicine, Department of Pathology, New York, NY, United States, 2 Medical Diagnostic Center, Yaoundé, Cameroon, 3 Manhattan Veterans Affairs Harbor Healthcare Systems, New York, NY, United States P34.2 LB Acute HIV- Subtype C Infection Is Associated with Rapid Increase of Tissue-like Memory and Decrease in Resting Memory B-cells Jennifer K. Mabuka,2, Anne-Sophie Dugast 3, Zelda Euler 3, Yathisha Ramlakhan 2, Krista Dong 3, Bruce D Walker 2,3,4, Thumbi Ndung u,2,3, Galit Alter 3 KwaZulu Natal Research Institute for TB & HIV, University of KwaZulu Natal, Durban, South Africa, 2 HIV Pathogenesis Programme, Durban, South Africa, 3 Ragon Institute of MGH, MIT and Harvard, Massachusetts General Hospital and Harvard Medical School, Boston, MA, United States, 4 Massachusetts General Hospital, Infectious Diseases, Boston, MA, United States Background: Superinfected HIV patients provide the unique opportunity to investigate the immune response after challenge with diverse HIV antigens and remain the major source for recombinant strain production. The competition and/or coexistence of two or more viral strains drive viral evolution and diversity, eventually triggering the generation of more broad and potent neutralizing antibodies (Abs) as compared to singly infected patients. The objective of this study was to analyze the genetic evolution of the viruses found within a superinfected Cameroonian patient, and to determine the neutralization sensitivity of the recombinant viruses to both autologous and heterologous neutralizing Abs. Methods: Longitudinal plasma samples from an HIV- superinfected Cameroonian patient were analyzed for their Env diversity, evolution, and recombination events. Pseudoviruses were generated from timepoints before, during, and after superinfection, and were then tested for sensitivity to homologous plasma and to heterologous neutralizing mabs. Results: Env sequence analysis indicated that the CRF02_AG infected subject became superinfected with an F2 strain resulting in a massive increase in viral diversity. Later, the patient s viral repertoire was narrowed to quasispecies closely clustering with the initial strain. The highest neutralization was observed with autologous plasma from timepoints after superinfection but before the intiation of ART. Of interest, the superinfecting F2 strain was resistant to neutralization with most known broadly neutralizing antibodies (bnabs) including PG09 and VRC0 while the original infecting CRF02_AG strain was sensitive to these bnabs. Conclusions: The resistance of the F2 strain to known bnabs urges the need to generate nabs from such patients and to monitor the emerging recombinant strains. HIV- superinfected patients may serve for the generation of new bnabs covering multiple subtypes and deliver important knowledge for future vaccine design. Background: HIV chronic infection (CI) is characterized by perturbations in B cell homeostasis, phenotype and function. There are limited data describing B cell dynamics during HIV acute infection (AI). Characterizing B cell subsets during AI might help define signatures that shape the humoral response in HIV infection. Methods: Eleven women who became HIV- infected during a longitudinal follow-up study in Durban, South Africa were analyzed. Samples were analyzed at baseline (pre-infection), at week, 2 week, month and 3 months post detection of plasma viremia. Multicolor flow cytometry was used to identify B cell subsets based on expression of CD2CD27 on live CD9+ lymphocytes and defined as follows; activated memory (AM) CD27+CD2-, resting memory (RM) CD27+ CD2+, naïve cells (N) CD27-CD2+, tissue-like memory (TLM) CD27-CD2- and plasmablasts (PBs) CD27+CD38+ cells. HIV-specific antibodies against subtype C gp20, gp4 and p24 antigens were determined by ELISA. Results: Compared to a baseline negative sample, we observed rapid and significant expansion of TLM cells post HIV infection (PI); week (p = ), 2 weeks (p = 0.008), month (p = 0.046) and 3 months (p= 0.043). In contrast, RM cells were significantly lower throughout AI compared to baseline; week (p = ), 2 weeks (p = 0.06), month (p = 0.09) and 3 months (p= 0.08). We observed significant expansion of AM cells at 2 weeks and month (p = 0.008, p= respectively) followed by contraction by 3 months (p = 0.260) PI. Peripheral blood PBs peaked by a median of 8 days (range 8-56 days) PI. There was a temporal relationship between peak viral load, PB peak and detection of HIV specific antibodies. Conclusions: Perturbations in B cell subsets occurs immediately following HIV- infection and this may therefore determine the subsequent development of anti-hiv antibodies

310 Posters Posters Posters 34: Glycans and Antibody Effector Functions P34.3 LB Potent SIV-specific Antibodies Targeting the Cyanovirin Binding Site Rosemarie Mason, Cameron Adams, Carole Bewley 2, John Mascola, Mario Roederer Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 2 Laboratory of Bioorganic Chemistry and Laboratory of Molecular Biology, The National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, United States Background: The SIV model of HIV infection is useful for studying vaccine mediated and immune correlates of protection but knowledge of the specificity, function and efficacy of protective SIV antibody responses is limited. Binding of cyanovirin-n (CVN), a potent inhibitor of HIV and SIV, occludes the CD4bs and 2G2 epitope - both major sites of HIV Env vulnerability. We used CVN with SIVgp40 to isolate CVNbsspecific mabs. Methods: Trimeric SIVgp40 and CVN proteins were used to generate 2 distinct probes with differential binding for CVNbs-specific B cells. Sequential staining with these probes was used to isolate CVNbs-specific B cells by indexed single cell sorting. Individual mabs were cloned and expressed in vitro and tested for SIV-specific binding and neutralization. Results: We sorted CVNbs-specific B cells from SIV-infected rhesus macaques. Of 5 mabs screened, bound to monomeric SIVgp20 and trimeric SIVgp40 but not SIV Env 5mer peptides. Pre-adsorption of SIVgp40 with CVN blocked binding of 4 out of SIV-specific mabs confirming their specificity for the CVNbs and validating our probe strategy. Interestingly, only the CVNbs-specific mabs (ITS50, ITS5, ITS52 & ITS53) neutralized SIVmac25.30 (Tier 2) with ITS50 and ITS5 also cross-neutralizing primary isolate HIV-2 (732A). Of note, ITS5 showed detectable albeit low level neutralization of SIVmac239. The remaining 7 mabs were binding but non-neutralizing. SIV CD4bs- and VV2- specific mabs did not compete with ITS50, ITS5 or ITS52 for binding to SIVgp40 trimer suggesting distinct and non-overlapping binding to SIV Env. Additional mapping with glycan-deficient SIVgp40 protein and individual glycan-deletion mutant viruses will determine binding specificity and glycan-dependence of these CVNbs-specific mabs. Conclusions: We isolated and characterized SIV CVNbs-specific mabs by a novel competitive binding probe strategy that may be adapted for isolating additional SIV- and HIV-specific mabs in order to optimize HIV vaccine development. 30 HIV Research for Prevention 204 HIV R4P

311 Posters 35: HIV Drug Resistance in vitro P35.0 Resistance Profile of the Diaryltriazine Nonnucleoside Reverse Transcriptase Inhibitor and Candidate Microbicide UAMC0398 Thursday, 30 October P35.02 Investigating HIV- Resistance to CCR5 Antagonist Maraviroc for the Design of New Prevention Strategies Kevin K. Ariën, Muthusamy Venkatraj 2, Johan Michiels, Katleen Vereecken, Jurgen Joossens 2, Pieter Van der Veken 2, Leo Heyndrickx, Jan Heeres 2, Koen Augustyns 2, Guido Vanham Institute of Tropical Medicine, Antwerpen, Belgium, 2 University of Antwerp, Antwerpen, Belgium Background: We previously selected UAMC0398 as a lead compound from a screen with 60 diaryltriazine analogues, in the context of a multipartner program on microbicide development (FP7-CHAARM). This new NNRTI has a superior toxicity profile compared to Dapivirine (DPV) and it retains nm activity against DPV-resistant viruses. We now report on the resistance profile of this new candidate microbicide. Methods: Resistance was induced in dose-escalation studies and in single high dose experiments. Mutations were identified by sequencing and subsequently confirmed in IC50 experiments with site-directed mutagenesis in a pnl4.3 molecular clone. Cross-resistance to other clinical and experimental NNRTIs was studied. Finally, the replication capacity of the UAMC0398-resistant viruses was assessed. Results: Dose-escalation studies revealed the following mutations in the RT gene: V90I, V06A, E38K, V79M, H22Y, F227C and M230I. Full blown resistance was selected only after 50 days. At least 4 of these mutations are required in concert for resistance against UAMC0398. Cross-resistance was assessed against DPV, Etravirine, Rilpivirine (RPV), Lersivirine, MIV70, Efavirenz and Nevirapine. Only Etravirine and RPV retained partial activity (sub µm). Single high dose exposure to UAMC0398 did not select for resistant HIV. Finally, we clearly show that UAMC0398-resistant viruses are significantly less fit than wild type virus. Conclusions: UAMC0398 is a strong new candidate microbicide with superior toxicity, activity against DPV-resistant HIV, and a complex resistance profile that is not easily selected but similar to RPV. Jacqueline K. Flynn,2, Michael Roche,2, Geza Paukovics, Hamid Salimi,2, Renee C. Duncan, Miranda S. Moore, Anne Ellet, Lachlan R. Gray,2, Becky Jubb 3, Mike Westby 3, Damian F. J. Purcell 4, Sharon R. Lewin,2, Benhur Lee 5, Richard J Payne 6, Melissa J. Churchill,2, Paul R. Gorry,2 Burnet Institute, Melbourne, Australia, 2 Monash University, Melbourne, Australia, 3 Pfizer Global Research and Development, Kent, United Kingdom, 4 University of Melbourne, Melbourne, Australia, 5 UCLA, Los Angeles, CA, United States, 6 University of Sydney, Sydney, Australia Background: Maraviroc (MVC) is a CCR5 antagonist currently used for the treatment of HIV- and is being tested as a Pre-exposure prophylaxis prevention strategy. MVC resistance can occur, however the mechanisms behind its development are unclear. Elucidating these mechanisms will assist in the design of improved CCR5 antagonists for use in prevention and therapy. Methods: Envs were cloned from plasma from two phase III MOTIVATE clinical trial participants, who developed phenotypically verified MVC resistance in vivo. Both participants were male with a similar duration of infection (4yrs). MVC resistance was characterised by measuring the maximal percent inhibition (MPI) and the ability of the Envs to recognise the MVC-bound confirmation of CCR5 (Affinofile assay). Tropism alterations for the infectivity of CD4 + T cell subsets and macrophages were characterised and CCR5 engagement of resistant Envs was tested for neutralization by sulfated peptide fragments of the CCR5 N-terminus. Results: The MPI values and affinity profiling showed that these Envs displayed a divergent ability to recognise MVC-bound CCR5, characterised by either a relatively efficient (MPI ~ 0%, less CCR5-dependant) or inefficient (MPI ~ 90%, more CCR5-dependant) recognition. Only the MVC-resistant Env with efficient recognition of MVC-bound CCR5 (MPI ~ 0%) displayed a tropism shift for CD4 + T cells characterized by a significant expansion of infected CM T cells, potentially affecting the HIV reservoir size and no change in macrophage infectivity. Interestingly, both resistant Envs were susceptible to neutralization by a sulphated peptide fragment of the CCR5 N-terminus. Conclusions: Our results suggest that the pattern of HIV tropism alterations for susceptible cells can vary depending on the magnitude of MVC resistance. Despite divergent phenotypes of resistance, both Envs showed an increase reliance on sulfated CCR5 N-terminus residues, which could provide a new avenue to block HIV- entry through CCR5 N-terminus sulfopeptidomimetic drugs. 3

312 Posters Posters 35: HIV Drug Resistance in vitro P35.03 Resistance Profile of CD4 Mimic Small Compounds (CD4MCs) and the Structure Analysis by Molecular Dynamic (MD) Simulation Shigeyoshi Harada, Masaru Yokoyama 2, Samatchaya Boonchawalit,3, Hironori Sato 2, Shuzo Matsushita 3, Kazuhisa Yoshimura,3 National Institute of Infectious Diseases, AIDS Research Center, Shinjuku-ku, Japan, 2 National Institute of Infectious Diseases, Pathogen Genomics Center, Musashimurayama, Japan, 3 Kumamoto University, Center for AIDS Research, Kumamoto, Japan P35.04 Combinations of Entry and Reverse Transcriptase Inhibitors as Candidate Microbicides Carolina Herrera, Natalia Olejniczak, Javier García Pérez 2, José Alcamí 2, Loïc Martin 3, Oliver Hartley 4, Charles Kelly 5, Robin Shattock Imperial College, Infectious Diseases, London, United Kingdom, 2 Instituto de Salud Carlos III, Madrid, Spain, 3 Comissariat à l Energie Atomique, Giff-sur-Yvette, France, 4 University of Geneva, Geneva, Switzerland, 5 King s College London, London, United Kingdom Background: CD4MCs inhibit the gp20-cd4 interaction and can also expose masked epitopes of neutralizing antibodies on the gp20 protein. In this study, we investigated the phenotypic change in the CD4MCs resistant isolates against CD4MCs, other entry inhibitors and anti-env neutralizing monoclonal antibodies (nmabs). Methods: Resistant variants were induced by five CD4MCs using the primary KP-5P virus (subtype B, R5) in PM cells. We constructed infectious clones with CD4MC-resistant mutation following in vitro selection. The susceptibility of the infectious clones to the inhibitors was evaluated using TZM-bl cells. We also simulated the gp20 3D structures by MD simulation model. Results: Resistance against CD4MCs was associated with V255M, T375N/I, or M426I substitutions. We examined susceptibilities of these mutated clones to the CD4MCs, maraviroc (MVC), an entry inhibitor IC9564, CD4bs nmab 3D6, and CD4i nmab 4E9C. V255M, T375I, and M426I were associated with high level of resistance to all CD4MCs tested, while there was no substantial difference between the wild type and the mutated clones in sensitivity of MVC and IC9564. The V255M and M426I clones became resistant to 4E9C, whereas the clone with T375I showed low sensitivity to both 3D6 and 4E9C. MD simulations of KP-5P gp20 in complex with NBD-556 showed that (i) V255M mutation abolished the interaction of NBD-556 and gp20, and (ii) M426I mutation disconnected a hydrogen bond between Lys30 and Glu429, thus the NBD-556 binding site shifted different from the usual. Conclusions: These data may give important knowledge for combination of NBD and other entry inhibitors or nmabs. Background: Multiple drug combinations as microbicides have been shown to be highly effective in preclinical studies against wild type HIV- isolates. This study aims to assess the activity of entry inhibitors (EIs) combinations with a nucleotide reverse transcriptase (RT) inhibitor (NRTI), against resistant HIV- and SIV isolates Methods: Antiviral efficacy of dual combinations of an NRTI, tenofovir (PMPA), and EIs, a CD4 mimetic miniprotein, M48-U, or CCR5 inhibitors, 5P2-RANTES or maraviroc; was evaluated. The combinations were assessed in cellular (TZM-bl cells and activated PBMCs) and colorectal explant models. Preincubation of cells or tissue with the drugs individually or in combination, for one hour was followed by addition of virus. NRTI-escape mutants with point mutations K65R +/- M84V in HIV-YU.2 and SIVmac32H RT were used. Infection was determined by measurement of luciferase expression (in TZM-bl cells) or p24/p27 viral antigen in culture supernatants Results: All PMPA-EI dual combinations inhibited the NRTI-resistant clones in all cellular and explant models tested. The dose-response curves of combinations including M48-U or 5P2-RANTES reflected the activity of the EI with no increase of potency of these drugs when combined with PMPA. The same result was observed with the gelformulated version of M48-U. Interestingly, an increase of activity was observed for maraviroc and PMPA when used in combination against all resistant isolates tested Conclusions: The positive results obtained against clade B NRTI-resistant HIV- isolates in this pre-clinical evaluation indicate that combinations of EIs with PMPA are good candidate microbicides able to block wildtype viruses and, importantly, NRTI-resistant isolates, which have been shown to be in increasing prevalence 32 HIV Research for Prevention 204 HIV R4P

313 Posters 36: HIV Incidence and Prevalence P36.0 Participation in Clinical Research Could Modify Background Risk for Trial Outcome Measures Andrew M. Abaasa, Gershim Asiki, Jonathan Levin, Ubaldo Bahemuka, Eugene Ruzagira, Freddie M. Kibengo, Jerry Mulondo, Juliet Ndibazza, Matthew A. Price 2, Pat Fast 2, Anatoli Kamali MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 2 International AIDS Vaccine Initiative, New York, NY, United States Thursday, 30 October P36.02 Trends of Reported HIV Sexual Risk Behaviour and HIV Incidence among Fisherfolk in Uganda Receiving Clinic-based Routine HIV Counseling and Testing Ubaldo Mushabe Bahemuka, Andrew Abaasa, Eugene Ruzagira, Freddie Mukasa Kibengo, Juliet Ndibazza, Gershim Asiki, Jerry Mulondo, Matthew Andrew Price 2, Patricia Fast 2, Anatoli Kamali Medical Research Council/Uganda Virus Research Institute Unit on AIDS, Entebbe, Uganda, 2 International AIDS Vaccine Initiative (IAVI), New York, NY, United States Background: Data on HIV incidence and retention are needed to inform study design of efficacy trials. However, the selection criteria and interventions during an actual clinical trial could reduce HIV incidence and thus affect the statistical power. We investigated the effect of inclusion and participation in a simulated vaccine efficacy trial (SiVET) on HIV and pregnancy incidence in a fisherfolk cohort in SW Uganda. Methods: High-risk vounteers aged 8-49 years from fishing communities 30-40km from the MRC/UVRI research centre were recruited in HIV open cohort. High risk was defined as history of multiple sex partners, unprotected sex, STI presence and absence from home for 2 days in the preceding 3 months. Consenting volunteers with at least 3 months of follow-up, no contraindications for hepatitis B vaccine and willing to use contraception were administered a licensed Hepatitis B vaccine at 0, and 6 months to mimic a candidate vaccine. The cohort was followed quarterly for a year. HIV incidence, pregnancy and retention rates were compared. Results: Of 853 (55% men) individuals screened from Jan 202-Feb 204, 575 (60% men, mean age 28) were enrolled into the open cohort, 282 (73% men) of whom enrolled into the SiVET between Jul 202-Feb 203. In both groups there was reduction of risky behaviours, (p< 0.05). A total of 3 HIV incident cases occurred in 93.0 PYO [brackets 95% CI]; incidence 3.9/00 PYO [8.-24.] and 0 cases in 3.6 PYO; incidence 3.2 [.7-6.0] in the open cohort and SiVET respectively. A total of 26 pregnancies were observed in 42.7 Women Years of Observation (WYO); incidence 60.9 [ ], and 4 pregnancies (7.4WYO); incidence 5.6 [2.-4.8] in the open cohort and SiVET respectively. Conclusions: Although reduction in risky sexual behaviours was observed in the open cohort and SiVET, lower HIV and pregnancy incidence rates were observed in the SiVET. The low HIV incidence could impact on sample size estimates for a prevention trial. Background: HIV counseling and testing (HCT) has been shown to reduce HIV risk behaviour and is central to HIV prevention programs. We investigated risk behaviour and HIV incidence trends in a fisherfolk cohort on Lake Victoria, Uganda. Methods: HIV negative volunteers aged 8-49 years, at high risk of HIV infection and willing to undergo HCT were enrolled. At every quarterly visit, they received HCT. Condoms and STI treatment were also provided. Risk behaviour data on alcohol consumption before sex, multiple or new sex partners, condom use and exchange of gifts for sex in the past 3 months were collected at baseline and every 6 months for 2 years. We fitted multilevel logistic regression models to investigate the trends. Results: A total of 428 (63% men) volunteers, mean age 28 years were enrolled. There were significant reductions in reported risk behaviours over the 2-year follow-up. The proportion reporting 2 partners decreased from 80% at baseline to 45% at month 6 and to 43% at month 24 for males; for females the decrease was from 42% at baseline to 3% at month 6 and to 6% at month 24; P< 0.0). Similarly there were significant reductions among men (P=0.0) reporting new partners but of borderline statistical significance among females (P=0.09). In both sexes there were significant decreases in reported non-condom use, transactional sex and in having sex when drunk. HIV incidence (in brackets 95% CI) reduced from 8.2/00 person years (5.-3.5), to 7.3 ( ), 6.5 (4.6-9.) and 6.0 ( ) at 6, 2, 8 and 24 months respectively (p=0.2). Conclusions: In this study there was a substantial reduction in selfreported risk behaviour in the first 6 months and marginal reduction in the later period. However, a modest HIV incidence reduction was observed. This calls for an urgent need for combination prevention strategies in this population. 33

314 Posters Posters 36: HIV Incidence and Prevalence P36.03 Development of a Risk Scoring Tool to Predict HIV- Acquisition in African Women Jennifer E. Balkus,2, Jingyang Zhang, Gonasagrie Nair 3, Thesla Palanee 4, Gita Ramjee 5, Clemensia Nakabiito 6, Marthinette Taljaard 7, Baningi Mkhize 8, Zvavahera Mike Chirenje 9, Jeanne M. Marrazzo 2, Elizabeth R. Brown,2, Barbra A. Richardson,2 Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 University of Washington, Seattle, WA, United States, 3 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa, 4 Wits Reproductive Health and HIV Institute, University of the Witwatersrand, Johannesburg, South Africa, 5 South African Medical Research Council, Durban, South Africa, 6 Makerere University - Johns Hopkins University Research Collaboration, Kampala, Uganda, 7 The Aurem Institute, Klerksdorp, South Africa, 8 Chris Hani Baragwanath Hospital, Johannesburg, South Africa, 9 UZ - UCSF, Harare, Zimbabwe P36.04 Age-disparate Partnerships and Risk of HIV- Acquisition among South African Women Participating in the VOICE Trial Jennifer E. Balkus,2, Gonasagrie Nair 3, Elizabeth Montgomery 4, Anu Mishra 2, Thesla Palanee 5, Gita Ramjee 6, Ravindre Panchia 7, Pearl Selepe 8, Barbra A. Richardson,2, Zvavahera Mike Chirenje 9, Jeanne M. Marrazzo 2 Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States, 2 University of Washington, Seattle, WA, United States, 3 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa, 4 RTI International, San Francisco, CA, United States, 5 Wits Reproductive Health and HIV Institute, University of the Witwatersrand, Johannesburg, South Africa, 6 South African Medical Research Council, Durban, South Africa, 7 Chris Hani Baragwanath Hospital, Johannesburg, South Africa, 8 The Aurem Institute, Klerksdorp, South Africa, 9 UZ - UCSF, Harare, Zimbabwe Background: In many African countries, women account for more than half of all new HIV- infections; however, not all women are at equal risk of acquiring HIV-. A risk prediction tool that can identify women at highest risk for HIV- acquisition could improve prevention research efficiency and inform HIV- prevention activities in policy and clinical settings. Methods: Using baseline data from VOICE (MTN-003), a randomized, double-blinded, placebo-controlled trial conducted in South Africa, Uganda and ZImbabwe that assessed safety and effectiveness of daily oral and vaginal chemoprophylaxis for HIV- prevention, we used standard methods for the development of clinical prediction rules to generate a risk scoring tool to predict HIV- acquisition over the course of one year. The predictive ability of the score was assessed by calculating area under the curve (AUC) and the score was internally validated using 0-fold cross-validation. Results: Among 5,029 women enrolled in VOICE, 4,834 women had complete data for factors of interest and were included in the analysis; of these, 248 acquired HIV- within one year after enrollment (HIV incidence=6.05% [248/4,093 person-years]). The final risk score resulting from multivariable modeling included the following baseline factors: participant age, married/living with a partner, financial or material support from partner, partner has other partners, curable STI, HSV-2 status and alcohol use. The maximum possible score was 2; 36% of participants had a score > 6 and accounted for 66% of HIV- infections. The AUC for the score was 0.72 and mean AUC from 0-fold cross validation was 0.70, indicating good predictive ability. Conclusions: A discrete set of characteristics which can be easily assessed were highly predictive of HIV- acquisition over one year. External validation of the risk score is required to evaluate the tool s performance when applied to different populations of women at risk for HIV- infection in Africa. Background: Age-disparate relationships where the male partner is older than the female partner have been associated with increased HIV acquisition risk in women. A recent analysis of data from South Africa failed to observe an association between age-disparate partnerships and HIV acquisition. We assessed the association between partner age and HIV acquisition among South African women in VOICE. Methods: VOICE was a randomized, double-blinded, placebo-controlled trial conducted at sites in South Africa that assessed the safety and effectiveness of daily oral and vaginal chemoprophylaxis for HIV prevention in women. Cox proportional hazards models stratified by site were used to assess participant-reported male partner age at enrollment and HIV acquisition risk in the first year of follow-up. Results: Among 4077 South African women enrolled, 3789 had complete data for this analysis. Of these, 26% and 5% reported having a male partner >5 and >0 years older at enrollment, respectively. There were 230 HIV infections within year of follow-up (38 personyears). Reporting a male partner >5 years older was not associated with HIV acquisition (HR.00; 95% CI 0.74,.35). Findings were similar for reporting a male partner >0 years older (HR 0.92; 95% CI 0.49,.74). Results for both male partner age categories were similar among younger and older women (women < 25 years: male >5 years, HR.0 [95% CI 0.7,.44]; male >0 years, HR.24; [95% CI 0.58, 2.66] versus women 25 years: male >5 years, HR 0.95 [95% CI 0.55,.65]; male >0 years, HR 0.68 [95% CI 0.2, 2.7]). Results were consistent after adjusting for known baseline risk factors for HIV acquisition in VOICE. Conclusions: These data corroborate recent reports and may suggest a shift in local epidemiology of heterosexual HIV transmission. Given the limitations of these analyses (use of enrollment data and inclusion of only one partner in analysis), regular assessment of characteristics to identify women at greatest risk of HIV acquisition is needed. 34 HIV Research for Prevention 204 HIV R4P

315 Posters 36: HIV Incidence and Prevalence P36.05 Estimating HIV Incidence for Identification of Microbicide Trial Sites in India: A Crosssectional Study Thursday, 30 October P36.06 Zambia FSW-risks Descriptors: HIV, Retention, Condom Use and Trichomonas Vaginalis/ Sperm Trends over Time Nomita Chandhiok, Ramesh S. Paranjape 2, Sanjay M. Mehendale 3, Archana Beri 2, Sanjay Chauhan 4, R Hari Kumar 5, Seema Sahay 2, Reynold Washington 6, Marianne Callahan 7 Indian Council of Medical Research, Division of Reproductive and Child Health, New Delhi, India, 2 National AIDS Research Institute, Pune, India, 3 National Institute of Epidemiology, Chennai, India, 4 National Institute for Research in Reproductive Health, Mumbai, India, 5 National Institute for Nutrition, Hyderabad, India, 6 Karnataka Health Promotion Trust, Bengaluru, India, 7 CONRAD, Eastern Virginia Medical School, Arlington, VA, United States Background: India has the second largest burden of HIV in the world with estimated 2. million infections. Of these, 39.3% are estimated to be women. The present study was designed to characterize HIV- incidence in select clinical site(s) suitable for conducting HIV prevention trials in India. Methods: This cross-sectional study was conducted in Female Sex Workers (FSWs) between Jan 202 and June 203 in six districts with historical evidence of concentrated HIV epidemic, from three high HIV prevalence states of India, namely Maharashtra, Andhra Pradesh and Karnataka. A total of 938 FSWs were enrolled. HIV incidence was estimated on sero-positive samples using serological assays such as BED-Capture Enzyme Immuno-assay (CEIA) and two avidity assays: modified GS HIV ½ (Biorad) and LAg HIV- (Sedia). In addition, molecular estimation was carried out on sero-negative samples using pooled PCR. Furthermore, HIV Incidence was calculated using Recent Infection Testing Algorithm (RITA) which was developed by including BED-CEIA, an avidity assay, and CD4 count in series. Results: While the HIV prevalence in this population was estimated at 9.26%, the incidence by serological assays ranged from 0.44 to 2.4. The False Recent Rates (FRR) for the two avidity assays were different; however, the incidence estimates were comparable. Similar HIV incidence was also obtained using RITA. HIV incidence by Pooled PCR was significantly different from serological assays. Conclusions: The incidence estimates varied with the test applied. Although there is no gold standard test available, two avidity based assays gave comparable results. Adding CD4 counts to RITA did not improve the outcome. The low HIV incidence estimates, obtained using serological assays in a cross-sectional study of FSWs in high prevalence states, suggest that efficacy trials for vaginal microbicides would require large sample sizes in India. Linda J. Kimaru, Tyronza Sharkey 2, Marydale Oppert 3, Kathleen Wu 2, Rachel Parker 3, William Kilembe 2, Mubiana Inambao, Amanda Tichacek 3, Susan Allen 3 Rwanda Zambia HIV Research Group, Ndola, Zambia, 2 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 3 Rwanda Zambia HIV Research Group, Atlanta, GA, United States Background: Female Sex Workers (FSWs) in Zambia are at high risk for HIV and may be eligible for HIV prevention clinical trials. HIV- FSWs were invited to enroll into a prospective cohort to determine the incidence and risk factors for HIV. Methods: From 202 to 204 FSWs in Lusaka and Ndola were invited for VCT/STI services from known FSWs hotspots through direct and peer FSW outreach activities. FSWs received HIV/STI testing and counseling and were offered Long Acting Reversible Contraceptives (LARC, IUD and implant). HIV- FSWs were invited for enrollment and follow-up visits quarterly for HIV/STI testing and counseling. Demographic and risk assessment questionnaires were done at each visit. Results: Among 733 FSWs screened, 39 (53%) were HIV-. Of the HIV-, 9% were positive for T. Vaginalis, 3% were positive for syphilis, 3% had presence of semen on their vaginal swabs and 9% were on LARC. The 297 (75%) HIV- FSW who returned for enrollment reported an average of 2 clients a month including and 2 repeat/regular clients. 7% reported condom use always/most of the time and 83% sometimes/ never. 48% reported non-condom use due to client request/refusal. 87 (64% of those enrolled) FSWs came for at least follow up visit, with a total of 803 months of follow up. initiated LARC. 32% reported disclosing their HIV status by client request and 60% had voluntarily disclosed. 64% had requested the HIV status of their clients. At followup 7% were positive for T. Vaginalis, 3% had presence of semen, and 70% reported not using a condom within the last month. Annual HIV incidence was 6%. Conclusions: HIV- FSW are at high risk for HIV/ STI infection and should be targeted for prevention trials. Though HIV- FSW received counseling, STI and LARC services, they maintained high-risk behaviors including unprotected sex. Disclosure of HIV- status may be dangerous for FSW if clients use this as an excuse to refuse condom use. Asking clients for their HIV status shows awareness but may not yield truthful information. 35

316 Posters Posters 36: HIV Incidence and Prevalence P36.07 Hematological Profiles of HIV-infected Adults Initiating Highly Active Antiretroviral Therapy (HAART) in Uganda Rachel Kyeyune, Elmar Saathoff 2, Amara Ezeamama 3, Wafaie Fawzi 4, Thomas Loescher 2, David Guwatudde 5 Infectious Diseases Institute, Research, Kampala, Uganda, 2 Medical Center of the University of Munich, Division of Infectious Diseases and Tropical Medicine, Munich, Germany, 3 University of Georgia, Epidemiology and Biostatistics, Athens, GA, United States, 4 Harvard School of Public Health, Nutrition, Boston, MA, United States, 5 Makerere University College of Health Sciences, School of Public Health, Epidemiology and Biostatistics, Kampala, Uganda P36.08 Characteristics of Clients Undergoing Repeat HIV Counseling and Testing Compared to Clients Newly Tested for HIV in Nyanza Province, Kenya Patrick O. Owiti, Kevin Owuor 2, Hillary Ng eno, Nicollate Awuor, Patricia Ong wen, Starley B Shade 3, Jayne Lewis-Kulzer 3, Elizabeth A Bukusi, Craig R. Cohen 3 Kenya Medical Research Institute, Family AIDS Care and Education Services, Kisumu, Kenya, 2 University of Reading, Statistical Services Center, Reading, United Kingdom, 3 University of California San Francisco, Obstetrics, Gynecology and Reproductive Sciences, Pediatrics, Medicine, San Francisco, CA, United States Background: Cytopenias are the most common HIV-associated hematological abnormality. Cytopenias become more prevalent as HIV progresses and are often fatal. Sex, race, geographical location and comorbidities such as tuberculosis have been associated with cytopenias. Data from resource-limited settings about the prevalence, correlates and trends in cytopenia are limited. This analysis assessed the prevalence and correlates of cytopenia at initiation of HAART and the trend in cytopenias among HAART-treated AIDS patients in Uganda. Methods: This is a secondary analysis of hematological data of 400 adults enrolled into the Multivitamins, HAART and HIV/AIDS Trial (NCT ). Anemia was defined according to WHO guidelines and leucopenia and thrombocytopenia were defined using study site laboratory reference ranges for lack of generally accepted standardized definitions for these 2 cell lines. Univariate and bivariate analyses were done to describe the patient population and log-binomial regression was used to quantify the correlates of cytopenia. Multilevel Mixed-effects linear regression was used to examine the change in the 3 cell lines over 8 months of HAART Results: Sixty five percent had at least one form of cytopenia and the prevalence was higher in females(pr.2 CI.0-.43) and higher with decreasing CD4 count and decreasing body mass index. Anemia was the most common occurring in 47.8%. Adjusted models showed that hemoglobin values were 0.03g/dl higher with each month of HAART (p< 0.00) while white blood cell counts and platelets were lower by 0.0(p=0.009) and 0.5 (p=0.522) units respectively with each month of HAART. Conclusions: Cytopenias are a frequent complication in HIV-infected adults at initiation of HAART in Uganda. Females, a decreasing CD4 count and decreasing body mass index were associated with having a cytopenia. This data shows that HAART improves hemoglobin status and alters the white blood cell and platelet counts independent of sex, immunological and nutritional parameters. Background: According to Kenya AIDS Indicator Survey 2007, only 35.6% of Kenyan adults had ever tested for HIV. Since then, routine HIV counseling and testing (HTC) has increased in health facilities. We compared characteristics of new and repeat clients tested for HIV to inform efforts to improve testing uptake. Methods: This retrospective study included a proportional stratified random sample of adult clients ( 8 years) tested at the outpatient department from October-December, 20 at 9 high patient volume facilities in Nyanza Province (collectively testing >2,000 clients in one quarter). Routine data were abstracted from health facility registers through systematic selection of every 5st adult patient. Variables included age, gender, HIV status, individual vs. couples testing, test type (new/repeat) and interval to repeat test. Descriptive statistics were presented as medians or proportions. Logistic regression was used to assess differences between new and repeat HIV testers. Results: Among the 555clients sampled, the median age was 27 years (IQR22-35), 365(66%) were female, 397(7%)were repeat testers and 52 (94%) tested as individuals. Median time to repeat test was 4 months (IQR3-7).New testers were older (aor=.36 per 0 year age increase; 95%CI.7-.58). HIV prevalence among new testers was higher than among repeat testers (27% vs. 3%, respectively) (aor2.64; 95% CI ).No significant gender differences were found between new and repeat testers (OR.3; 95 % CI ) or individual vs. couple visit (OR.2; 95% CI ). Conclusions: The majority of adult patients seeking HIV testing have been tested previously. Those newly tested for HIV have a higher HIV prevalence than repeat testers. This may indicate that initial testing is reaching higher risk individuals and that HTC along with other preventive interventions may have led to lower HIV incidence amongst repeat testers. Further attention to behavioural differences between new and repeat testers should be examined. 36 HIV Research for Prevention 204 HIV R4P

317 Posters 36: HIV Incidence and Prevalence P36.09 ABSTRACT WITHDRAWN Thursday, 30 October P36.0 HIV and STI Incidence and the Association with Number of Lifetime Sexual Partners - Case for Combination HIV/STI Prevention Strategies Renee A. Street, Neetha Morar, Handan Wand 2, Gita Ramjee South African Medical Research Council, Durban, South Africa, 2 Kirby Institute, University of New South Wales, Sydney, Australia Background: South Africa has a generalised HIV epidemic driven largely by heterosexual transmission. Multiple sexual partnerships is believed to be an important driver of the HIV epidemic. The aim of this study is to describe socio-demographic characteristics of women by the number of lifetime sexual partners and its association with incident HIV and sexually transmitted infections (STI). Methods: The Methods for Improving Reproductive Health in Africa (MIRA) clinical trial was conducted between 2003 and In Durban (KwaZulu-Natal Province), a total of 485 women were enrolled from two sights (a peri-urban clinic in Umkomaas and a rural clinic in Botha s Hill) and were followed up for a total of 24 months. The chi squared test was used to compare categorical parameters. Kaplan-Meier survival analyses were carried out to estimate the crude HIV and STI incidence rates over time. All analyses were performed using Stata V.0.0 (College Station) and SAS V. 9.2 Results: Women with a greater number of lifetime sexual partners (5+) were older (35+ years of age), had early sexual debut (< 6 years of age) and were unmarried (p< 0.00). Cohabitation status and level of education were not significantly associated with lifetime number of sexual partners. Women with 5+ lifetime sexual partners had a crude HIV incidence reported at 0.5 per 00 person years (PY). Women with 3 lifetime sexual partners were at higher risk of STI acquisition (25 per 00 PY) when compared with women with lifetime sexual partner (4 per 00 PY). Conclusions: This study supports existing evidence that reducing partner turnover is key in HIV prevention. However combination HIV/ STI prevention strategies are essential to achieve maximum impact on HIV prevention. Women remain critical participants in investigative biomedical and translational HIV prevention efforts. 37

318 Posters Posters 36: HIV Incidence and Prevalence P36. High HIV Incidence in Young Men who Have Sex with Men Engaged in Sex Parties, Factors Associated with Sex Party, Bangkok MSM Cohort Study, Warunee Thienkrua, Sarika Pattanasin, Tareerat Chemnasiri, Anchalee Varangrat, Wichuda Sukwicha, Supaporn Chaikummao, Sumetha Hengprasert, Anupong Chitwarakorn 2, Timothy H. Holtz,3 Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Department of Diseases Control, Ministry of Public Health, Nonthaburi, Thailand, 3 Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and Prevention, Atlanta, GA, United States Background: Evidence concerning sex party participation and recreational drug use along with continuing high rates of HIV and STIs among MSM in Thailand has been described in several studies. We investigated HIV incidence among young MSM (YMSM) and characteristics associated with sex party participation. Methods: Thai men, 8 years from the Bangkok metropolitan area who reported sex with another man in the past 6 months were enrolled and followed-up every four months for HIV testing and audio computerassisted self-interview behavioral questionnaire completion. We defined engaging in sex parties as having group sex and using recreational or erectile dysfunction drugs in the past four months preceding enrollment. We calculated HIV incidence among YMSM 8-24 years of age from using survival analysis. Factors associated with sex party participation were analyzed using logistic regression. Results: Of 744 men enrolled, 72 (40.8%) were YMSM, 77/72 (%) reported engaging in sex parties, 428/72 (60%) reported unprotected anal intercourse in the past 4 months. HIV incidence among YMSM engaged in sex parties was 8 per 00 Person-Years. Factors associated with sex parties were: sex with casual partners at pub/disco [Adjusted Odd Ratio (AOR) 5.8, 95% CI ], paid for sex (AOR 4., 95% CI.9-8.8), sex with foreigner (AOR 3.3, 95% CI.9-5.8), self-report of STIs (AOR 2.8, 95% CI.5-5.), had HIV infection (AOR 2.6, 95% CI.5-4.7), reporting Internet use to find casual partners (AOR 2.0, 95% CI.-3.5), ever had suicidal idea (AOR 2., 95% CI.2-3.7), and experience of coercive sex (AOR.9, 95% CI ). Conclusions: Engaging in sex parties was common among Bangkok YMSM in our cohort. Sex at entertainment venues and commercial sex are strongly associated with sex parties, both high-risk behaviors for incident HIV. Innovative and creative HIV interventions directed at MSM engaging in high-risk behaviors is needed to promote safer sex behaviors. 38 HIV Research for Prevention 204 HIV R4P

319 Posters 37: Immunogenetics P37.0 Variants in Vitamin D Pathway and Antiviral Response Genes Interact to Modulate the Natural Resistance to HIV- Infection Thursday, 30 October P37.02 The Expression Analysis of Hexokinase Gene in the Pumwani Commercial Sex Worker Cohort, Nairobi, Kenya Wbeimar Aguilar-Jiménez, Wildeman Zapata,2, Antonio Caruz 3, Joan Fibla 4, Marina Laplana 4, Antonio Rivero 5, Juan A. Pineda 6, Maria T. Rugeles Grupo Inmunovirología, Universidad de Antioquia UdeA, Medellin, Colombia, 2 Grupo Infettare. Universidad Cooperativa de Colombia, Medellin, Colombia, 3 Unidad de Inmunogenética, Facultad de Ciencias Experimentales, Universidad de Jaén, Jaen, Spain, 4 Unitat de Genètica Humana, Departament de Ciències Mèdiques Bàsiques, IRBLleida, Universitat de Lleida, Lleida, Spain, 5 Maimonides Institute for Research in Biomedicine of Cordoba (IMIBIC)/Reina Sofia University Hospital, Cordoba, Spain, 6 Unidad Clínica de Enfermedades Infecciosas y Microbiología. Hospital Universitario de Valme, Sevilla, Spain Background: The immunomodulatory functions of vitamin D (VitD) may orchestrate anti-hiv- responses and influence resistance to HIV-- infection exhibited by HIV--exposed seronegative (HESN) individuals. Methods: We performed a nested case-control study, involving HIV- -exposed seropositives (cases) and seronegatives (controls), from two cohorts: sexually-exposed from Colombia, and parenterally-exposed from Spain. The association of 40 variants in 9 genes of the VitD pathway, and in 3 genes of the antiviral response with resistance/ susceptibility (R/S) to HIV- was evaluated. Results: Twenty-four variants were associated with R/S to HIV- infection at p< 0.05 (including 6 variants at false discovery rate [FDR] 20%). Similarly, 2 haplotypes in both pathways were also associated with R/S to HIV- after Bonferroni correction. Some variants in VitD pathway genes displayed epistatic interactions with those in antiviral genes and such interactions were associated with R/S to HIV- infection at p< 0.0, but not after Bonferroni correction. Furthermore, most of the HESNs exhibited the same combinations of the variants in VitD pathway and antiviral genes (p< after Bonferroni correction). Remarkably, we observed that HESN individuals carrying resistance-associated variants in genes coding for the vitamin D receptor (VDR), 27-OH hydroxylase (CYP27A), and Toll-like receptor 2 (TLR2), had higher levels of VitD in plasma, of VDR mrna in blood leucocytes and mrna of beta-defensins in mucosa, suggesting that the expression of these molecules could be genetically determined. Interestingly, variants in antimicrobial peptides that exert their function on mucosal surfaces were associated with R/S in sexually- but not in parenterally-exposed individuals, highlighting differences in mechanisms underlying resistance to HIV-, depending on the route of exposure. Conclusions: These results suggest that the VitD pathway may act in concert with antiviral genes modulating the resistance phenotype observed in HESN individuals. Winnie Apidi, Ruey Chi Su, Joshua Kimani,2, Frank Plummer,2,3, Blake Ball,2,3 University of Manitoba, Medical Microbiology & Infectious Diseases, Winnipeg, MB, Canada, 2 University of Nairobi, Nairobi, Kenya, 3 Public Health Agency of Canada, National Microbiology Laboratory, Winnipeg, MB, Canada Background: Altered susceptibility to HIV- infection has been observed in a commercial sex worker (CSW) cohort in Nairobi, Kenya, where a subset of women are classified as HIV- exposed yet seronegative (HESN). A gene expression analysis conducted showed differential regulation of the glycolysis/gluconeogenesis pathway in HESN CSWs. The first and potentially critical regulatory step in glucose metabolism is its entry into lymphocytes, where glucose binds to Hexokinase- that prevents it from leaking out. Together with the Glucose Transporter, Hexokinase- regulates the first rate-limiting steps of the entire glucose metabolism by phosphorylating glucose into glucose-6-phosphate, which is the starting material for glycolysis. Methods: The study population was randomly selected from the Pumwani Sex Worker Cohort, Nairobi including: HIV highly exposed yet seronegative (HESNs) CSWs (>7 years); newly enrolled HIV-uninfected (< 3 years) CSWs, 85% of whom would likely seroconvert to HIV- positive; and lowly-exposed HIV negative antenatal clinic attendees with low exposure to HIV. Total RNA was extracted from PBMCs using Trizol; cellular Hexokinase- mrna levels were quantified by quantitative real time PCR using SYBR Green. Statistical analysis was performed using Mann-Whitney U Test. Differences were considered to be significant if P< Each assay was normalized using 8s rrna gene. Results: We observed significantly lower level of Hexokinase- mrna expression in HESNs when compared to that in newly enrolled HIV uninfected CSWs. (Hexokinase- p=0.0323) Furthermore, the levels of Hexokinase- mrna in HESN and the HIV negative antenatal clinic attendees were quite similar. (Hexokinase- p=0.6448) Conclusions: Lower expression of Hexokinase- in HESN might suggest lower regulation of glucose uptake. Hexokinase is a rate-limiting enzyme in the glycolysis pathway. Following studies of expression and uptake are underway to understand its role in glucose metabolism in HIV resistance. 39

320 Posters Posters 37: Immunogenetics P37.03 P37.04 Subtype-Specific HIV- Adaptation to Host HLA Guinevere Q. Lee, Jonathan Carlson 2, Chanson J. Brumme, Helen Byakwaga 3,4, Conrad Muzoora 3, Daniel MacMillan 5, Natalie Kinloch 5, Kyle Cobarrubias 5, Mark A. Brockman,5, Peter W. Hunt 4, Jeff N. Martin 4, Mary Carrington 6, David R. Bangsberg 7, P. Richard Harrigan,8, Zabrina L. Brumme,5 BC Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada, 2 Microsoft Research, Seattle, WA, United States, 3 Mbarara University of Science and Technology, Mbarara, Uganda, 4 University of California San Francisco, San Francisco, CA, United States, 5 Simon Fraser University, Faculty of Health Sciences, Burnaby, BC, Canada, 6 Frederick National Laboratory for Cancer Research, Cancer and Inflammation Program, Laboratory of Experimental Immunology, SAIC Frederick, Inc., Frederick, MD, United States, 7 Massachusetts General Hospital and Harvard University, Boston, MA, United States, 8 University of British Columbia, Department of Medicine, Vancouver, BC, Canada Background: HIV- adapts to HLA alleles expressed by infected hosts and host populations. However, the extent to which HIV s genetic context influences population-level adaptation to HLA remains incompletely understood. The Ugandan HIV epidemic, where subtypes A and D co-circulate, provides a unique opportunity to distinguish universal pathways of HIV adaptation from those that are subtype-specific. Methods: High-resolution HLA class I and HIV RNA gag genotyping were performed for 53 antiretroviral-naïve patients from Kampala and Mbarara, Uganda. Recombinant and non-a/d sequences were excluded, leaving 200 subtype A and 35 subtype D gag sequences for analysis. HLA-associated polymorphisms were identified in each subtype via statistical association with phylogenetic correction, after which a logistic regression approach was used to determine cases where the same HLA allele drove significantly different escape pathways between subtypes. Results: Of 03 unique HLA alleles observed in the study cohort, only 3 (B*58:0, C*6:02, A*26:2) differed in frequency between subtypes A and D (p< 0.05), consistent with a single host population where HIV subtypes co-circulate. A total of 55 HLA-associated polymorphisms at 25 Gag codons were identified at the population level in subtype A; 36 HLA-associated polymorphisms at 25 Gag codons were identified in subtype D (p< 3x0-4, q< 0.2). Comparative analysis revealed that >35% of these adaptations differed significantly between subtypes. For example, B*57:03 drove the selection of T242N in subtype D (Odds Ratio~250, p=2x0-0 ) but not in subtype A (inter-subtype comparison p=8x0-6 ), whereas Y79F selection by A*0:0 was six-fold stronger in subtype A (OR~20) versus D (p=2x0-3 ). Conclusions: HIV s genetic context exerts a substantial influence on its ability to adapt to HLA. Establishing whether this is attributable to differential epitope presentation, mutational constraints or other factors is relevant to vaccine design. Characterization of the 3 Untranslated Region of HLA-G in HIV- Infected Black South African Mothers and their Infants Heather A. Hong,2, Maria Paximadis,2, Glenda E. Gray 3, Louise Kuhn 4, Caroline T. Tiemessen,2 Centre for HIV & STIs, National Institute for Communicable Diseases, NHLS, Johannesburg, South Africa, 2 Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 3 Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital, Soweto, South Africa, 4 Gertrude H. Sergievsky Centre, College of Physicians and Surgeons, Mailman School of Public Health, Columbia University, Department of Epidemiology, New York, NY, United States Background: HLA-G is a tolerogenic molecule capable of inhibiting the cytolytic activity of natural killer cells and cytotoxic CD8 T-cells. A 4bp insertion-deletion (indel) within the 3 untranslated region (3 UTR) has been found to alter HLA-G expression, wherein high expression was associated with the Del/Del genotype. Other single-nucleotide polymorphisms (SNPs) within the 3 UTR have also been reported to influence HLA-G expression by binding several micrornas. Recently, the 4bp indel has been associated with susceptibility to vertical transmission of HIV-; however the roles of the other 3 UTR SNPs have not been extensively investigated. Methods: We sequenced the 3 UTR of HLA-G in a total of 26 Black South African HIV- infected mother-infant pairs as well as 7 HIVnegative controls. Mother-infant pairs were classified as HIV- nontransmitting (NT, n=44) or HIV- transmitting (TR, n=72). Results: We identified 8 previously reported polymorphisms arranged in 8 distinct haplotypes. Heterozygosity at three of the polymorphisms (the 4bp indel, +300C/G and +342G/C SNP) were significantly underrepresented in HIV-positive mothers compared to HIV-negative controls (P=0.09, P=0.00 and P=0.00, respectively). SNPs +300C/G and +342G/C were in complete linkage disequilibrium, and the +342G/C SNP was predicted to bind to three mirnas (mir-48a, mir-48b, and mir-52). There were no significant differences in variants of the 3 UTR between NT and TR mothers, or their exposed-uninfected and HIVinfected infants. Conclusions: This is the first description of HLA-G 3 UTR variability in a Black South African population. The data suggest that, in our cohort, variations within the 3 UTR of HLA-G did not influence vertical transmission. However, significantly higher heterozygous representation for the 4bp indel and +342G/C SNP in HIV-negative controls as compared to HIV-infected individuals, suggests a fine balance in HLA-G expression may be necessary for protection against sexual acquisition of HIV HIV Research for Prevention 204 HIV R4P

321 Posters 37: Immunogenetics P37.05 Fcγ Receptor Variability in the South African Population - Will this Impact on HVTN097 Vaccine Efficacy? Ria Lassauniere,2, Caroline T. Tiemessen,2 National Institute for Communicable Diseases, Centre for HIV and STI s, Johannesburg, South Africa, 2 University of the Witwatersrand, Faculty of Health Sciences, Johannesburg, South Africa Thursday, 30 October P37.06 A Potential Role for CXCR6 in Long-term Nonprogression of HIV- Infected Black South African Individuals Anabela C. P. Picton, Maria Paximadis, Neil Martinson 2, Caroline T. Tiemessen Centre for HIV and STIs, National Institute for Communicable Diseases, NHLS and Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 2 Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital, Soweto, South Africa Background: Vaccine-induced IgG binding antibodies interacting with Fcγ receptors (FcγR) may reduce infection risk through antibodydependent cellular cytotoxicity, triggering of soluble antiviral factors, or phagocytosis. The potential role of FcγR genetic variation in vaccine efficacy (VE) was previously demonstrated by the significant association of the FcγRIIc-T8I variant with VE in RV44 vaccinees. With the RV44 follow up trial (HVTN097) commencing in Southern Africa, we considered it important to establish the FcγR genetic variability in South Africans, given that this may influence VE. Methods: We genotyped FcγR functional variants and gene copy number in 3 South African Black individuals. The following variants were assessed: FcγRIIa-p.H3R, FcγRIIb-p.I232T, FcγRIIc-p.T8I, FcγRIIIa-p.F58V, FcγRIIIb-HNAa/HNAb/HNAc, FCGR2B/C promoter variants, and genetic markers predicting FcγRIIc expression. The FcγR data from South African Black individuals were compared with that of the 000 Genomes Project for Asians, European Caucasians and other African populations. Results: The FcγRIIc-8I allele previously associated with VE was not detected in the 3 South African Black individuals. Furthermore, none were predicted to express the FcγRIIc protein. Genotype frequencies were significantly different when comparing South African Black individuals with Asians (FcγRIIa-p.H3R, P < 0.000; and FcγRIIIa-p. F58V, P < 0.0), or with European Caucasians (FcγRIIb-p.I232T, P < 0.000; and FcγRIIIa-p.F58V, P 0.00). Of note were the significantly different genotype frequencies between African populations (South African, Kenyan, Nigerian) for FcγRIIa-p.H3R, FcγRIIb-p.I232T, and FcγRIIIa-p.F58V, suggesting differing FcγR-mediated immune function capabilities. Conclusions: The HVTN097 trial will commence in different African populations, who are characterized by substantial inter-population genetic diversity, which may differentially influence VE in the populations most in need of an effective HIV vaccine. Background: A role for the HIV secondary coreceptor, CXCR6, in long-term nonprogression (LTNP) to AIDS has been reported. A single nucleotide polymorphism (SNP), rs , located 42bp downstream from the CXCR6 termination codon, significantly associates with LTNP but not elite control (EC) in cohorts of European descent. In this study we investigated the role of rs in HIV- disease progression in populations of sub-saharan descent. Methods: Study participants included HIV--infected Black South African individuals with differing disease phenotypes (progressors, n=09; LTNPs, n=47; ECs, n=) and healthy HIV-uninfected Caucasian (n=28) and Black (n=36) individuals. A continuous region encompassing the CXCR6 open reading frame (ORF) and the untranslated regions (7.0kb) was amplified in 4 overlapping sections. Amplified fragments were sequenced and analysed for the presence of SNPs, indels and intragenic haplotypes to identify potential rs associated haplotypes. Realtime PCR assays were developed to detect the rs SNP and another SNP, CXCR6-E3K, also shown to associate with clinical outcome of HIV- infected individuals. Results: The rs SNP frequency did not differ between LTNPs (5.%) and progressing (56.9%) Black individuals (P=0.386). CXCR6 gene sequencing failed to detect linkage disequilibrium between rs and other SNPs within CXCR6. CXCR6-E3K, located in the ORF, was found to be present at high allelic frequencies within control Black individuals (44.4%) and absent in the Caucasian individuals genotyped. In LTNPs heterozygosity for the E3K SNP was overrepresented (72.3%; 85.3% of whom also had rs ) compared to progressing (34.9%) individuals (P< 0.00, OR=0.7). Conclusions: The E3K SNP is likely to affect both CXCR6 cell surface expression and/or binding of its ligand. Thus, we hypothesize that the high prevalence and interplay of both these mutations in Black South African individuals may be masking the effect of the rs SNP on LTNP within this population. 32

322 Posters Posters 37: Immunogenetics P37.07 P37.08 Cytokines Genes Polymorphisms in Ukrainian HIV- Infected Individuals Anna Ivanivna Piddubna Sumy State University, Infectious Diseases and Epidemiology Department, Sumy, Ukraine Background: The objective of the research was to study distribution character of the allelic variants of cytokines genes in HIV- infected Ukrainians. Methods: Data for the study were DNA samples, received from 200 inhabitants of Ukraine: 78 HIV-infected, 22 - HIV-negative individuals from the group of high risk of contamination, 00 healthy blood donors. IL-4 (-590C/T), IL-0 (-592C/A) and TNF-α (-308G/A) genes polymorphisms detection was made with PCR-RLFP method. Results: By analysis of frequency of IL-4 gene allelic variants it has been discovered that homozygotes by the main allel were the dominant variant. Among people with HIV T/T minor gene carriers were 4.5 more often met in comparison with control group (p< 0.05) that can prove the tendency to association of the mentioned genotype with infection. Distribution of allelic variants of IL-0 gene promoter region in position -592 is characterized by homozygote dominance by the main gene. Among the individuals with HIV A/A minor allel carriers were 3.4 more often met in comparison with control group (p< 0.05). Individuals with A/A genotype were not identified in group of high risk of virus infection. The abovementioned proves the tendency to association of minor allel carrier state with HIV infection. The occurrence of the homozygous combination of the allelic variant G/G of the promoter of TNF-α has been shown to prevail almost twofold over the occurrence of the variant G/A among all groups. High frequency of heterozygote by the main allel has been recorded among the individuals with HIV. Thus, G/A genotype frequency in group of HIV-infected people 2 and.5 exceeded the appropriate indices of group of high risk of infection and comparison group correspondingly (p< 0.05) that points to the tendency to association of the mentioned variant with infection. Conclusions: Cytokines genes variations may contribute to the acquisition of HIV infection and encourages carrying out of further populations studies in this sphere of HIV-infection immunogenetics. Multiple T-cell Epitopes of HIV- Nef Containing Positively Selected Mutations Associated with Different Disease Outcome Elnaz Shadabi, Raghavan Sampathkumar, John Ho 2, David La 2, Rupert Capina 2, Binhu Liang 2, Jeff Tuff 2, Joshua Kimani 3, T. Blake Ball,2, Francis Plummer,2, Ma Luo,2 University of Manitoba, Department of Medical Microbiology, Winnipeg, MB, Canada, 2 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada, 3 University of Nairobi, Nairobi, Kenya Background: HIV- Nef plays a major role in enhancing the pathogenicity of the virus through various mechanisms such as downregulation of CD4 and HLA class I surface expression and interfering with cell signaling pathways. Identifying and characterizing CD8+ T cell epitopes in Nef that are under host immune selection can help in selecting targets for an effective vaccine. Methods: 326 subtype A Nef sequences from treatment naïve patients of a Kenyan sex-worker cohort were generated using 454 pyrosequencing. Positively selected (PS) mutations were determined using a bioinformatics approach, quasi analysis. Peptides were designed with mutation placed in anchor position 2, 5, 8, 9 of epitopes of HLA class I alleles for validation with ELISpot assay using patient PBMCs. Results: E70D, I09V and I76M were associated with rapid CD4 decline (p=0.00, 0.05, respectively). H24N and K90M were associated with slow CD4 decline (p=0.00 and 0.029). The five PS mutations were significantly associated with HLA class I alleles including A*23:0 (E70D, p=0.002; I76M, p=0.003), A*02:0 (I09V, p=0.028; H24N, p=0.02), B*58:0 (I09V, p=0.048), A*3002, B*57:03 and C*02:0 (H24N, p=0.026, , and 0.0 respectively) and C*06:02 (K90M, p=0.037). ELISpot analysis identified 27 novel epitopes containing either the consensus or the PS mutations. Six new epitopes contained E70D, five epitopes contained K90M, and I09V and H24N were each contained by eight new epitopes. No epitopes containing I76M was confirmed by ELISpot. It is possible that I76M represents compensatory mutations due to functional requirements under host immune selective pressure. Conclusions: Identification and characterization of epitopes containing beneficial and detrimental PS mutations can provide important insight for selecting immunogens for an effective HIV vaccine. More detailed investigation of T-cell responses, such as poly-functionality and proliferation to these mutations will be conducted to further characterize these Nef epitopes. 322 HIV Research for Prevention 204 HIV R4P

323 Thursday, 30 October Posters 38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring P38.0 P38.02 Operational Challenges for the Set-up of Gram Stain Analysis for Diagnosing Bacterial Vaginosis in a Local Laboratory in Durban, South Africa Natasha Gounden, Nathlee Abbai, Rashika Maharaj Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: Studies have demonstrated the association between BV and HIV acquisition in woman. The prevalence of BV reported in clinical trials conducted by the Medical Research Council HIV Prevention Research Unit (MRC HPRU) is between 5-9%. In an attempt to reduce turnaround time for results, enable direct contact with research clinics, facilitate staff capacitation and reduce costs, the HPRU was selected by the Protocol Reference Laboratory to perform in-house testing for diagnosing BV. We report here on the operational challenges associated with the set-up of the laboratory to perform the analysis. Methods: To perform the testing, the laboratory underwent infrastructural changes in order to implement a staining workbench. Correct waste management disposal was also part of the set-up process as well as procurement for staining reagents, consumables and internal quality control (IQC) slides. Staff that were involved in the testing had to undergo rigorous training. Results: Operational challenges included: training and evaluating only staff who specialize in clinical pathology and microbiology, quality of gram stain reagents, QC processes, waste management and manual IQC slides which were compared to commercially available ones. Commercial gram stain reagents and IQC slides were preferred as less labour intensive with enhanced staining results. Waste receptacles proved futile and a local municipal waste discharge permit was obtained instead. To date, 605 slides have been analyzed. The following errors were noted during the QC process: incorrect slide preparation (47%) and inadequate sample material for diagnosis (0.3%), however, retraining of clinical staff resulted in a significant reduction (50%) of errors. Conclusions: Despite operational challenges HPRU has been successful in the set-up of gram stain analysis for BV diagnosis as a Protocol Reference Laboratory. This enables BV to be diagnosed in real-time rather than shipping to an international laboratory where slides are analyzed at study end. Impact of Tenofovir % Gel on Hepatitis B Virus Resistance in CAPRISA 004 Cheryl Baxter, Sinaye Ngcapu, Jason T. Blackard 2, Eleanor A. Powell 2, Patricia K. Penton 2, Quarraisha Abdool Karim,3, Salim Abdool Karim,3 Centre for the AIDS Programme of Research in South Africa, Durban, South Africa, 2 College of Medicine, University of Cincinnati, Division of Digestive Diseases, Department of Internal Medicine, Cincinnati, OH, United States, 3 Columbia University Mailman School of Public Health, Department of Epidemiology, New York, NY, United States Background: The intermittent, before-and-after sex, dosing regimen used in the CAPRISA 004 trial, where women were only required to use the gel on days that they had sex, meant that women in the trial who were infected with Hepatitis B Virus (HBV) could be exposed to variable levels of a single antiretroviral agent in the presence of HBV, potentially increasing the risk for the development of antiviral resistance. Here we assessed the impact of intermittent tenofovir gel use on HBV resistance. Methods: Hepatitis B virus DNA was extracted from stored samples of women identified as being HBV surface antigen positive during the trial. Extracted DNA was amplified by polymerase chain reaction (PCR) and positive samples were gel purified and sequenced using the Applied Biosystems 330 x; automated sequencer. Five of the samples were cloned into the pgem-t Easy vector. Sequences were submitted to the Stanford University HBV drug resistance database for analysis of drug resistance, resistance loci and resistance-associated mutations. To confirm conventional PCR results, the same HBV DNA was then amplified by nested PCR and tested for resistance using the commercial INNO-LiPA HBV DR V3 kit. Results: A 3.2 kb PCR product corresponding to the pol region of HBV was successfully amplified in 9/37 (5%) of the β-globin positive samples, 3 from women assigned to tenofovir and 6 from women assigned to placebo. All samples clustered phylogenetically with HBV subtype A. There was no difference in the frequency of amino acid variation between the tenofovir and placebo arms. None of the known tenofovir resistance mutations (M240V/I, L80M, A94T, V24A, N238T) were identified using both the commercial kit and in-house PCR assay. Conclusions: No resistance mutations associated with tenofovir were detected in the 9 sequences examined. These data provide further support for the safety of intermittent % tenofovir gel use as HIV preexposure prophylaxis for HBV-infected individuals

324 Posters Posters 38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring P38.03 P38.04 Setting up and Oversight of Local Clinical Trial Site Laboratories with Limited Resources: A Case Study of South Africa s FACTS 00 Study Sarah S. Cohen, Ishana Naidoo, Romeo Martin 2, Nomfundo Maduna 2, Maletsatsi Moloelang 3, Nwabisa Ndzamela 3, Christian Kasango 4, Andrew Tlagadi 5, Gail Stockenstrom 6, Nomzamo P. Tabata 6, Thuleleni Dlungwana 7, Siphiwe Gumede 7, Manyabeane A. Phaahla 8, Zaheda Ismail 9, Elizabeth Rammutla 0, Sinead Delany-Moretlwe, Glenda Gray, Helen Rees, Lindiwe Nhlangulela 4 Wits Reproductive Health and HIV Institute, University of the Witwatersrand, Research, Johannesburg, South Africa, 2 Wits Reproductive Health and HIV Institute (WRHI), Research, Johannesburg, South Africa, 3 Perinatal HIV Research Unit, Kliptown - FACTS, Soweto, South Africa, 4 Aurum Institute for Health Research, Laboratory, Rustenburg, South Africa, 5 Aurum Institute for Health Research, Laboratory, Tembisa, South Africa, 6 Desmond Tutu HIV Foundation, Laboratory, Cape Town, South Africa, 7 MatCH Research (Maternal, Adolescent and Child Health Research), Laboratory, Pietermaritzburg, South Africa, 8 Medunsa Clinical Research Unit, Laboratory, Garankuwa, South Africa, 9 Qhakaza Mbokodo Research Clinic, Laboratory, Ladysmith, South Africa, 0 Setshaba Research Centre, Laboratory, Soshanguve, South Africa, Perinatal HIV Research Unit, Chris Hani Baragwanath Hospital, Research, Soweto, South Africa Background: Ensuring Good Clinical Laboratory Practice (GCLP) in low-resource field laboratories sites during clinical trials is challenging. We describe processes for ensuring GCLP compliance at all nine South African sites in the FACTS 00 trial. Methods: All nine sites established laboratories for collection, processing and shipping of samples to a central laboratory; three sites also processed and stored onsite. A coordinating team developed a study-wide lab analytical manual (LAM), checklist for lab assessments, and a laboratory working group that met monthly by phone to discuss issues. Regular onsite assessments of laboratories were completed at six-weekly intervals to ensure conformance with protocol and GCLP. All visit findings were recorded in reports which were reviewed and assessed for trends in findings over this period. Results: An average of 0 assessments per site was completed in the period July 202 to May 204. Monthly lab working group calls provide opportunities for sites to resolve findings, and discuss common challenges and retraining needs.interventions for improvement included retraining of personnel, vertical audits of sample records, and provision of guidance in documenting laboratory incidents and deviations. Following this, improvement was noted in general, and fewer deviations were identified suggesting that the interventions were successful at improving overall laboratory quality assurance. Conclusions: While audits help ensure compliance with protocol requirements, regular supportive supervision to sites helps identify areas of deviation sooner and allows for rapid intervention and correction of potential problems. Clinical Research Site Preparedness for Clinical Emergencies - Implications for Preexposure Prophylaxis, Microbicide and Vaccine Trials Tashni Nayager, Vaneshree Govender South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa Background: The frequency of clinical emergencies is far lower in HIV prevention trial sites versus medical healthcare facilities. Despite this, patients with urgent and emergent conditions such as cardiac arrest, acute asthma, seizures, anaphylaxis, hypoglycaemia and shock, must be provided with immediate care to decrease morbidity and mortality. Our aim was to evaluate if emergency trolleys were appropriately stocked, if staff were adequately trained in clinical emergency management and if systems for monitoring adequacy were sufficient. Methods: At the HIV Prevention Research Unit (HPRU), the primary study population over the last decade has been healthy 8-45 year old women, with a smaller population of infants and HIV sero-converters. The stringent screening eligibility process excludes individuals with serious comorbidities, such as unstable cardiovascular disease. Emergency trolley equipment, supplies and resuscitation medication were evaluated for availability, expiry dates and functional acceptability with monitoring checklists. Results: An internal audit of 6 clinical research sites identified a need for a consistent schedule for trolley checks, timeous ordering of new stock, regular maintenance of oxygen cylinders, review of required medication, easy access to medication in a temperature controlled environment, easy access to resuscitation algorithms and a study operational procedure. Due to the infrequent occurrence of medical emergencies, staff competency was assured with refresher training. Appropriate systems were in place for immediate referral to a Department of Health care facility based on a Memorandum of Understanding. Conclusions: A clinical emergency management plan is a necessity in a research site conducting HIV prevention trials. The level of emergency care provision should be tailored to the study and site requirements, and be relevant to the study population. Safety of clinical trial participants is of the utmost importance. 324 HIV Research for Prevention 204 HIV R4P

325 Thursday, 30 October Posters 38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring P38.05 P38.06 Introduction of a Novel Monitoring Tool to Reduce Specimen Archive Errors Lakshmi Jagesur, Rashika Maharaj, Avika Haridutt, Nathlee Abbai, Duduzile Ndwandwe, Gita Ramjee,2 Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 London School of Hygiene & Tropical Medicine, Epidemiology and Population Health, London, United Kingdom Background: The Medical Research Council (MRC), HIV Prevention Research Unit (HPRU) has been involved in the conduct of multiple HIV prevention clinical trials. In order to efficiently track clinical samples from the 6 Clinical Research Sites (CRSs), the laboratory used the Laboratory Data Management System (LDMS) for specimen archive. However, previous specimen verification quality error reports had identified gaps from the collection to archive process within the LDMS system. Therefore, the laboratory developed a novel monitoring tool as part of the quality specimen management programme. Methods: The monitoring tool was set up for: quality control checks of LDMS documentation against physical specimen verification and monitoring of monthly quality trends. The tool captured the following information: type of errors identified per CRS; participant identifiers (PTIDS); staff responsible for errors, Corrective Actions and Preventative Actions (CAPA) and re-trains performed. Results: For the period of September 202-December 203, out of samples archived, there were 80 errors: LDMS typographical errors (26/80), incorrect collection dates (9/80), incorrect visit codes (47/80), incorrect Case Report Form (CRF) completion (27/80), incorrect tracking sheet completion (3/80) and CRF and LDMS discrepancies (58/80). The number of errors on average per month ranged from 7 to 5. The new monitoring tool enabled stringent monitoring of specimen management with a significant decrease in errors to a total of 4 errors in December 203. Conclusions: Since the introduction of the award winning novel monitoring tool an improvement to MRC specimen archive quality has been noted. We recommend that the tool can be adapted by other organizations to improve their long term and high volume specimen archive processes. Comparison of 2 Different BD PROBETEC ET Extraction Methods for Detecting of Chlamydia trachomatis and Neisseria gonorrhoeae in HIV Prevention Trials Rashika Maharaj, Nathlee Abbai, Gita Ramjee,2, Lakshmi Jagesur Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom Background: Nucleic acid amplification tests for the detection of C. trachomatis (CT) and N. gonorrhoeae (NG) in genital tract specimens has become the standard diagnostic method used in most laboratories for HIV prevention trials. The BD ProbeTec ET System, use Strand Displacement Amplification (SDA) technology for the direct, qualitative detection of CT and NG from DNA extracted from endocervical swabs, male urethral swabs, and urine specimens. The aim of this study is to compare two DNA extraction methods (M) (M as per package insert; M2 an international developed in house method) for the detection of CT and NG using the BDProbeTec ET instrument. Methods: Our sample size included 60 vaginal swabs. The first set of extractions (method ) was performed by conventional lysing, priming and amplification. The second method (2) included additional washing and centrifugation steps prior to lysis, priming and amplification. During the amplification process, external quality controls [College of American Pathologists: CAP ] were included in the runs. As part of the validation process, samples were also processed at an external/reference laboratory. Results: There was 00% concordance for CT/GC between the results obtained by the external/reference laboratory and MRC HPRU Central Laboratory using method, 00 % sensitvity and specificity on GC/CT for all 60 swab samples. Method 2 achieved a 78 % concordance; CT had 6 false negatives =70% sensitivity; 85 % specificity; while GC had 4 false negatives=80% sensitivity; 90 % specificity. CAP panels achieved 00% pass on both methods. Conclusions: Method as per package insert was superior to method 2 achieving a higher sensitivity and specificity. It is vital in populations with high HIV risk with co-infections with CT and NG that methods used for detection are accurate, specific and in keeping with the package insert which this study illustrates

326 Posters Posters 38: Innovations in Vaccine and Microbicides Studies in Lab and Monitoring P38.07 P38.08 Hematologic and Chemistry Normal Laboratory Values among Healthy Ugandan Women Screened for a Pre-exposure Prophylaxis Trial: the MTN-003(VOICE) Study Flavia Matovu Kiweewa, Holly M. Gundacker 2, Mike Mubiru, Betty Kamira, Brenda Mirembe Gati, David Ojok 3, Samuel Kabwigu, Philippa Musoke,4, Clemensia Nakabiito, Mary Glenn Fowler,5 MU-JHU Research Collaboration, Kampala, Uganda, 2 SCHARP-FHCRC, Seattle, WA, United States, 3 MU-JHU Core Laboratory, Kampala, Uganda, 4 Makerere University College of Health Sciences, Department of Pediatrics and Child Health, Kampala, Uganda, 5 Johns Hopkins University, Baltimore, MD, United States Background: Universal laboratory toxicity grading tables are used to screen and monitor adverse event (AE)s in clinical trials for HIV prevention and treatment; use of these tables excludes otherwise eligible volunteers and can make AE assessment and product management challenging in specific populations. We computed selected hematologic and chemistry normal ranges specific to healthy non-hiv infected women screened for an HIV prevention trial (MTN-003) and compared findings to U.S. established intervals. Methods: Excluding women with hepatitis, syphilis, and pregnancy, we analyzed data from 538 women aged 8-45 screened in Kampala from Nov 2009-Dec 20. We calculated 95% normal intervals as the 2.5% and 97.5% limits for the population, and compared data against U.S.- derived lab intervals from Massachusetts General Hospital and Division of AIDS (DAIDS), Dec 2004 toxicity tables (clarification dated Aug 2009) to determine the number of women with any AE per DAIDS grading criteria. Results: Compared to intervals from the U.S., we found slightly lower 2.5 th centiles for red cell indices (hemoglobin (Hgb), hematocrit, MCV, red blood cell counts), lower white blood cells and neutrophils, but higher eosinophils. Chemistry parameters were comparable with U.S.- based ranges except for a lower 2.5 th centile for serum phosphorus and higher 97.5 th centile for ALT and AST. When graded against U.S.-derived DAIDS criteria, we observed 96 AEs from 87 (6%) volunteers with grade results including decreased neutrophils and phosphorus in 48 (9%) and 23 (4%) women respectively. There were 8 (%) grade 3 and no grade 4 AEs. Conclusions: Similar to existing local intervals, we found differences in upper and lower ranges for some hematologic and chemistry indices among healthy Ugandan women compared to US norms. About in 6 women were described as having grade toxicity for either Hgb, white cell indices, or phosphorus. Local laboratory ranges should be considered for toxicity grading in international research settings. Strategies to Optimise Data Quality Metrics in the ASPIRE Trial at the Wits Reproductive Health and HIV Institute in Johannesburg Pranitha Ramchuran, Krishnaveni Reddy, Helen Rees, Thesla Palanee Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: Quality data collection and their timeous reporting are critical in clinical trials. In ASPIRE, a phase 3 safety and effectiveness study of the Dapivirine Vaginal Ring in HIV prevention, data quality is assessed by the quality control (QC) error rate calculated by the number of errors per 00 case report forms (CRFs) submitted to the Data Management Centre (DMC). Data timeliness is defined as the percentage of CRFs received at the Data Management Centre (DMC) within 7 days of the visit. Methods: To ensure good quality data and timeliness at Wits RHI, several strategies were implemented that include the formation of a multi-disciplinary quality management (QM) team comprising Clinical Quality Improvement Mentors (CQIMs), Quality Assurance (QA)/QC officers and a datafax team. The CQIMs conduct clinical review and immediate retraining of the clinical team, QA/QC officers conduct error trend analyses and the site datafax team conducts a final QC prior to datafax. Other strategies include regular review and revision of QM processes within the Clinical Quality Management Plan; timely re-training of the team on errors trends identified and collective team review and ownership of monitoring and audit findings. An additional tool assisting this process is idatafax; a DMC data system that provides early access to errors identified on datafaxed CRFs. Its use facilitates identification of errors trends more frequently to inform improved CRF completion and shorter QC resolution times. Results: Implementation of these strategies has led to the development of a more structured and focused QM team and instilled a culture of accountability among the study team who consider quality in all aspects of trial implementation. Conclusions: These strategies are continuously reviewed and amended to suit the needs of the study and the dynamic nature of data collection and with consistent implementation have and continue to assist the team to strive to optimize data quality and timeliness metrics in ASPIRE. 326 HIV Research for Prevention 204 HIV R4P

327 Posters 39: Molecular Epidemiology P39.0 Using Viral Dynamics to Connect Clinical Markers of Disease Progression to Sequence Evolution during HIV Infection Thursday, 30 October P39.02 Adaptation of HIV- Envelope Glycoprotein gp20 to Humoral Immunity over the Course of the Epidemic Andrew E. Adams, Zabrina L. Brumme 2, Alexander R. Rutherford, Ralf W. Wittenberg Simon Fraser University, Mathematics, Burnaby, BC, Canada, 2 Simon Fraser University, Faculty of Health Sciences, Burnaby, BC, Canada Background: Understanding how HIV establishes infection and, if left untreated, eventually overcomes the immune system is crucial to the development of a vaccine or cure. The high rates of turnover and evolutionary adaptability exhibited by HIV pose a particular challenge to HIV vaccine development. Our focus is to understand the dynamics of two of the most commonly tracked clinical markers of an HIV infection: CD4+ T cells/mm 3 (CD4 count) and HIV RNA/ml (viral load). Methods: We developed a stochastic system of differential equations to model HIV infection that uses equilibration, adaptation, and inheritance to model the initial infection as well as successive generations of viral lineages. The model allows viruses to generate new lineages in proportion to their viral load with an inherited fitness. These lineages compete for immune cells to infect and drive decline in CD4 count through a series of small adaptations. We use this model to demonstrate how viruses with a sufficiently high mutation rate could overcome the immune system, even when most changes are expected to be detrimental to viral fitness. Results: We have calibrated our model to match viral load set points and rates of CD4 decline from 9 HIV-infected individuals studied longitudinally during early stages of the disease. Our model demonstrates how a genetically diverse population of viruses could be sustained in an environment with high rates of competition, turnover, and the development of an immune response. The underlying stochastic process also generates a phylogenetic structure which can be used to explore different hereditary patterns in the underlying viral lineages. Conclusions: Our model demonstrates that the high rate of mutation and recombination in the HIV genome can contribute to slow disease progression. It suggests the diversity of HIV lineages is a consequence of lineages having similar fitness and the high levels of competition which create a balance in the expansion of existing lineages and their replacement by new lineages. Melanie Bouvin-Pley, Marion Morgand, Alain Moreau, Laurence Meyer 2,3, Cécile Goujard 2,3, Hugo Mouquet 4, Michel C. Nussenzweig 5, Craig S. Pace 6, David D. Ho 6, Pamela J. Bjorkman 7, Daniel Baty 8, Patrick Chames 8, Marie Pancera 9, Peter D. Kwong 9, Pascal Poignard 0, Francis Barin,, Martine Braibant INSERM U 966, Tours, France, 2 CESP INSERM U 08, Paris, France, 3 AP-HP, Hôpital de Bicêtre, Le Kremlin-Bicêtre, France, 4 Institut Pasteur, Paris, France, 5 Howard Hughes Medical Institute (HHMI), Rockefeller University, New York, NY, United States, 6 Aaron Diamond AIDS Research Center, Rockefeller University, New York, NY, United States, 7 California Institute of Technology, Pasadena, CA, United States, 8 CRCM INSERM U 068, Marseille, France, 9 Vaccine Research Center, NIH, Bethesda, MD, United States, 0 International AIDS Vaccine Initiative (IAVI), Neutralizing Antibody Center, Scripps Research Institute, Immunology and Microbial Science, La Jolla, CA, United States, Laboratoire de Bactériologie-Virologie, CHU Bretonneau, Tours, France Background: Since 2009, a large panel of broad and potent monoclonal neutralizing antibodies (MoNAbs) against HIV- have been isolated. These MoNAbs can protect from HIV- infection and suppress established infection in animal models. Because their efficacy should be evaluated in human clinical trials, it is of importance to define the sensitivity of the most contemporary transmitted variants to these MoNAbs. We, and others previously, reported that HIV- has become more resistant to neutralization over the course of the epidemic (Bunnik et al, Nature Med 200, Bouvin-Pley et al, PloS Pathog 203). Methods: Here we extended the analyses to the most potent MoNAbs described since then, either more recently isolated or improved by structure-based gene modifications. Results: We fully confirmed the first observations showing an increasing resistance of HIV- clade B over time to MoNAbs targeting the major gp20 epitopes but not to MoNAbs targeting the gp4 MPER. Despite this evolution, some MoNAbs still were able to neutralize efficiently the most recently transmitted HIV- variants ( ). The most potent MoNAbs were the bi-specific PG9- and PG6-iMab that alone were able to neutralize all variants at less than 0.4 mg/ml. The sensitivity to imab remained similar over time, suggesting that the trend of increasing resistance to PG9-/PG6-iMAb may be attributed only to the antigen binding domain of PG9/PG6. NIH45-46m2 (and -m7), and 0E8 were also highly potent and, if combined, reached the potency of PG9-/PG6-iMAb. We also observed that 3BNC7 was almost as potent as the modified NIH45-46 antibodies, and that the lama-derived JM4IgG2b was the most potent Ab among those that do not target the major gp20 neutralizing epitopes. Conclusions: These data clearly suggest a continuous drift of the env gene of HIV- clade B over the epidemic, and that not a single epitope is concerned but the entire gp20 as a whole. The consequences of this adaptation on the envelope functionality are being explored

328 Posters Posters 39: Molecular Epidemiology P39.03 P39.04 Characterisation of Transmitted and Non-transmitted HIV in Index-recipient Transmission Pairs Lotte Bracke, Elisabeth Willems, Astrid Gall 2, Paul Kellam 2, Sandra Coppens, Conor Meehan 3, Georgios Pollakis 4,5, Ben Berkhout 4, Guido Vanham, Marion Cornelissen 4, Leo Heyndrickx, Kevin Ariën Institute of Tropical Medicine, Department of Biomedical Sciences, Virology Unit, Antwerp, Belgium, 2 Wellcome Trust Sanger Institute, Cambridge, United Kingdom, 3 Institute of Tropical Medicine, Department of Biomedical Sciences, Mycobacteriology Unit, Antwerp, Belgium, 4 Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands, 5 Institute of Infection and Global Health/CIMI, University of Liverpool, Liverpool, United Kingdom Background: Many of the viral and host factors associated with HIV transmission are still poorly understood. In 60-80% of the mucosal infections, a single transmitted/founder virus is responsible for the establishment of a productive infection, indicating a strong genetic bottleneck upon transmission. We aim to better understand the viral factors involved during transmission by studying the genetic variability and replicative characteristics of viruses isolated from transmission pairs. Methods: We had access to blood samples from 5 index-recipient transmission pairs of MSM. All samples were obtained shortly after transmission. Plasma was used for full genome sequencing and PBMC were cocultured with HIV negative donor PBMC by limiting dilution to obtain biological clones. Results: We isolated a total of 270 biological clones from the 5 transmission pairs. Nearly the complete gp20 from 7-8 clones was sequenced for each of the 0 individuals. As expected, these sequences group nicely with the plasma sequences. In only one transmission pair studied, an identical clone was found in both index and recipient. Phylogenetic analysis showed a low genetic diversity in the recipients, in contrast to a greater genetic diversity among the clones from the index patients. Sequences from recipients also showed shorter V, V2 and V4 loops, indicating a more compact envelope compared to viruses from the index patients. We also found fewer potential N-linked glycosylation sites in three recipients compared to their indexes. Conclusions: Our sequences and phylogenetic analysis confirm observations from others. Currently the replication capacity of the biological clones is assessed in dual infection/competition assays. This will allow us to rank the fitness of the clones obtained from all the transmission pairs. A selection of clones will also be tested against neutralizing antibodies and entry inhibitors. These experiments should give us an indication of which virus characteristics favour transmission. Significance of HIV- Western Blot Bands Appearance in Clinical Trials - Point of Seroconversion and Window Period in Rural Kwazulu-Natal; South Africa Rashika Maharaj, Nathlee Abbai, Gita Ramjee,2, Lakshmi Jagesur Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom Background: Information on acutely HIV- infected individuals is very useful for treatment, pathogenesis and disease progression. Besides accurately predicting HIV- infection, sequential appearance of specific bands of the western blot offers a window of opportunity to develop a less subjective tool for monitoring disease progression. The aim of this analysis is to investigate the significance and order of HIV- band appearance at two time points during seroconversion in HIV Prevention trials. Methods: Between July 2007 and December 202 the Medical Research Council (MRC) HIV Prevention Research Unit (HPRU) Central Laboratory did a retrospective analysis on Biorad Genetics HIV- Western Blot test performed on 652 plasma samples collected at two time points i.e. point of sero-conversion (T, n=326) and -2 post sero-conversion (T2, n=326). All testing was performed according to the manufactures instructions. Results: Of the 326 at T/T2: presence of HIV- Gp 60 (#/%) =303(93%) /324(99%), Gp 20 =82 (56%) /255(78%), Gp 65 =59(49%)/ 25(77%), p55/5=306(94%)/ 324(99%), Gp 4= 68(52%)/ 250(77%), p40=304 (93%)/ 323(99%), p3=95(60%)/ 276(85%), p24=32(96%)/ 323(99%), p8=32(4%)/ 89(58%). p24 antigen was the first marker to be detected within 2 weeks of infection evident in 99% of the samples tested. Antibodies to the envelope glycoproteins (gp 60 and gp 20) and the trans membrane glycoprotein (gp 4) appeared within 2 to 4 weeks of infection. Samples at both time points showed that Gp60, p55/5, p40 and p24 were predominantly evident in > 93-99% of all acute sero-convertor cases with majority displaying at least four of the nine bands characteristic of the virus by Western blotting, with the lowest number of three bands characteristic of the virus displayed by any sample. Conclusions: The use of WB banding patterns during early infection will prove useful as there is a renewed interest for new surveillance technologies in this area and may assist in future vaccine development 328 HIV Research for Prevention 204 HIV R4P

329 Posters 39: Molecular Epidemiology P39.05 Changes in Viral Population Kinetics Following HIV- Superinfection Murray G. Logan, Daniel J. Sheward, Colin Anthony, Nigel Garret 2, Carolyn Williamson University of Cape Town, Clinical Laboratory Sciences, Cape Town, South Africa, 2 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa Thursday, 30 October P39.06 The Role of N-glycosylation in DC-SIGN Interactions with Transmitted Founder Variants of HIV- Subtype C Envelope Evelyn N. Lumngwena, Liliwe Shuping, Netanya Bertniz, Claudia Cicala 2, James Arthos 2, Zenda Woodman University of Cape Town, Molecular and Cell Biology, Cape Town, South Africa, 2 National Institutes of Health, Laboratory of Immunoregulation, NIAID, Bethesda, MD, United States Background: Elucidation of factors influencing HIV superinfection with a second viral strain, despite pre-existing immune responses to the primary infecting strain, may provide insights into correlates of protection. Superinfection is frequently associated with a spike in viral load which could be due to loss of control of the primary infecting virus, or the superinfecting virus, or both. To evaluate if pre-existing responses to primary infection differentially controlled viral populations following superinfection, we estimated viral population kinetics at time points before and after superinfection. Methods: We performed 454 deep sequencing of two genomic regions, gag p7 (332bp) and env C2C3 (403bp), on three participants who were known to be superinfected. To enable quantification of the input cdna, and control for PCR and sequencing errors, the cdna was labelled with a unique primer ID. Results: In all three participants the temporary increase in viral load associated with superinfection was predominantly due to the superinfecting viral strain, which became the major circulating variant in two of the three participants. Within the regions examined, we did not detect recombination in the gag region, however in one of three participants recombination was found within the env C2C3 region in ~20% of viral sequences 5 weeks after superinfection. Conclusions: The superinfecting virus was not controlled following infection within two participants, suggesting that responses elicited to the primary strain were not cross-protective against the second strain at the time of superinfection. In one participant however, there was subsequent control of the superinfecting strain following its introduction, this could be indicative of a broader CTL response and warrants further investigation. Background: The design of effective vaccines and microbicides requires understanding early steps in mucosal transmission. As the genetic bottleneck at transmission favors variants with fewer potential N-glycosylation sites (PNGs) and shorter variable loops, we investigated whether Envelope (Env) PNGs could influence the ability of variants to cross the genital mucosa by altering gp20 interactions with DC-SIGN, which could favour transfer to CD4+ cells, and also alter localized immune responses in the genital epithelium. Methods: We initially determined whether pseudovirus-envs from transmitted founders (TF) had enhanced DC-SIGN binding, transinfection and increased IL-0 secretion over matched chronic Envs. As DC-SIGN interactions with Env favors high mannose N-glycans, we also deleted gp20 PNGs bearing high mannose glycans either singly, or in combination, in matched CAP239 Env T/F and chronic clones. Results: T/F Envs induced more IL-0 secretion than chronic controls. When PNGs were deleted from the CAP239 envs, the effect on pseudovirion entry, DC-SIGN binding and trans-infection was clone specific, suggesting that specific N-glycans affect Env function differently in different clones. For example deletion of PNG 448 reduced entry efficiency, DC-SIGN binding and trans-infection of TF by ~50% when compared to wild type, while either enhancing or maintaining these phenotypes in the chronic infection clone. Only deletion of the PNG 24 reduced IL-0 induction for T/F clones. Conclusions: As pseudovirion entry efficiencies of most PNG mutants were reduced for both CAP239 Env clones, it is difficult to determine the role that each might play in DC-SIGN interactions. However, the TF Envs induced MDDCs to secrete higher levels of IL-0 compared to matched chronic infection controls, suggesting that localized anti-inflammatory responses in the genital epithelium might play a role in HIV transmission

330 Posters Posters 39: Molecular Epidemiology P39.07 Real Time Fitness Assay of Two CRF0_A/E HIV- Transmitted Founder Variants Melanie Merbah, Gustavo Kijak, Leigh Anne Eller, Eric Sanders- Buell, Brendan T. Mann, Devin M. Pillis, Anne Marie O Sullivan, Meera Bose, Jenica L. Lee, Kultida Poltavee, Nelson Michael, Jerome Kim, Merlin Robb, Sodsai Tovanabutra, Agnes-Laurence Chenine MHRP/WRAIR/HJF, Silver Spring, MD, United States P39.08 Outgrowth of Subtype C Envelope Viral Populations in Dual-infected Individuals Is Not Always Associated with Entry Efficiency Shatha Sultan Ahmed Omar, Daniel Sheward 2, Carolyn Williamson 2, Zenda Woodman University of Cape Town, Molecular and Cell Biology, Cape Town, South Africa, 2 University of Cape Town, IIDMM, Cape Town, South Africa Background: RV27/ECHO study presents a unique opportunity to identify subjects very early in HIV infection, with a median time to last negative nucleic acid test (NAT) of 4 days. Two transmitted/founder (T/F) variants were isolated from a Thai volunteer, with a representation of 99% and % for the major (Mj) and the minor (mn) variants respectively at the peak of viral load. Using next-generation, targeted deep sequencing, we confirmed the presence of that mn variant on the day of the first positive NAT. After peak viremia the mn variant grew exponentially, reaching >30% by day 3. Six months post infection, the mn variant became the predominant quasispecies. We hypothesized that viral fitness is responsible for this viral dynamic profile. Methods: Full-length infectious molecular clones (FLIMC) of the Mj and mn variants were generated. Each Mj and mn FLIMC was engineered to express the fluorescent proteins, GFP and mcherry. The four constructs Mj.C2, Mj.G2, mn.c2 and mn.g2 were tested for infectivity in cell lines (TZMbl & A3R5) and primary cells (PBMC, macrophages, dendritic cells) and monitored by flow cytometry, fluorescent microscopy and p24 capture assay; cells were singly infected and co-infected. Cell tropism was also assessed using CXCR4 and CCR5 Ghost cells. Results: Both mn and Mj FLIMCs showed productive infection in cell lines and primary cells and used the co-receptor CCR5 exclusively. Neither the GFP nor mcherry proteins modified in vitro infection kinetics of Mj and mn FLIMCs. Cell lines and primary cells were co-infected with different ratios of Mj/mn. Contrary to their representation at the time of initial infection, we found that the mn variant was more fit, as measured by replication kinetics, than the majority transimitted Mj variant. Interestingly, dual infection of single cells was a very rare event. Conclusions: We successfully applied a new fitness assay to evaluate multiple T/F in acute infection and found fitness didn t account for dominance of the Mj variant during acute infection. Background: Dual infection with two phylogenetically distinct HIV variants has been reported to be associated with rapid disease progression possibly due to the emergence of early recombinant viruses with high viral fitness resulting in high viral loads. As it has previously been shown that Envelope (Env) plays an important role in HIV fitness, this study aimed to determine the relationship between in vivo viral outgrowth and Env entry efficiency, and to identify fitness determinants that can be used as novel targets for drug and vaccine design. Methods: Highlighter plots of SGA-derived env sequences of four dual infected individuals (sampled at enrolment and approximately 3, 6, and 2 months post infection (mpi) was used to determine the relative frequency and fluctuation of invivo viral populations within each participant. Representative amplicons were cloned and compared using a pseudivirion entry efficiency assay. Results: our data indicated the presence of recombinants at enrolment for 3/4 participants and the outgrowth of these viruses at 2 mpi for all participants suggesting rapid recombination. However, this rapid recombination was not always associated with enhanced entry efficiency as recombinants at 2 mpi had similar entry efficiency to those at the earliest time point for 2/4. In one participant (CAP84) recombination occurred within the signal peptide and gp4, with the recombinant carrying an additional N-glycan site (PNG) at position 339 in gp20. While the recombinant entered TZM-bl cells as efficiently as the primary virus, a chimeric Env carrying the gp4 of the recombinant but lacking the PNG had a 4-fold increase in entry efficiency. When this PNG (N339) was introduced by site directed mutagenesis, entry efficiency decreased suggesting that recombination did select for a fitter variant but that the introduction of an N-glycan lowered fitness. Conclusions: Therefore, viral outgrowth in HIV- dual infected individuals is likely due to a balance between replicative fitness and immune escape. 330 HIV Research for Prevention 204 HIV R4P

331 Posters 39: Molecular Epidemiology P39.09 Thursday, 30 October P39.0 HIV-2/SIV Vpx Protein Interacts with Human Nup53 and Regulates Viral Pathogenesis Satya Prakash Singh, Pankaj Gupta, S Mahalingam Indian Institute of Technology Madras, Biotechnology, Chennai, India Background: Vpx, a virus associated accessory protein is encoded by Human Immunodeficiency Virus type 2 (HIV-2) and Simian Immunodeficiency Virus (SIVsm/SIVmac lineage) is known to be involved in the nuclear import of viral DNA in non-dividing primary target. Interestingly, Vpx mutant virus fails to replicate in non-dividing cells. The mechanism by which Vpx helps in the nuclear transport of viral genome remains unknown. Methods: Co-immunoprecipitation was performed to study the proteinprotein interaction in transiently transfected HEK cells. Protein colocalization was assessed by immunofluoresence. Using homology modeling of Vpx and Nup53 amino acid sequences, putative binding motif for Vpx in Nup53 was predicted. Site directed mutagenesis was employed to generate mutant viruses which were defective for interaction with Nup 53. Nuclear import ability of wild type and mutant virus was analysed by 2-LTR assay. Results: Molecular transport across nuclear envelope is governed by nucleo pore complexes (NPCs), composed of 30 nucleoporins (NUPs). We found that interaction between Vpx and human Nup53 was necessary for viral DNA import. We mapped the domain of Nup53 critical for interaction with Vpx and our data suggests the role of a zinc finger domain (60-869aa) to be critical. Vpx interaction with Nup53 was impaired by exchange of serine (63,65) and tyrosine (66, 69 and 7) residues to alanine and resulted in abrogation of nuclear localization. Interestingly, the SIV PBj.9 with mutant Vpx was found to have reduced nuclear import ability in 2-LTR assay. Conclusions: Our data gives insight into the mechanism of nuclear import by Vpx interaction with Nup53. Novel treatment methods using phosphorylation inhibitors specific for Vpx could be devised in future. The Sexually Driven Epidemic in Youngsters of China s Southwestern Border Region Was Caused by Dynamic Emerging Multiple Recombinant HIV- Strains Huamian Wei, Jenny H. Hsi, Song Duan 2, Yi Feng, Cui He, Yuhua Ruan, Xiang He 3, Lingjie Liao, Yanling Ma 4, Yunda Huang 5, Manhong Jia 4, Hui Xing, Yiming Shao State Key Laboratory for Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China, 2 Dehong Center for Disease Control and Prevention, Dehong, China, 3 Guangdong Provincial Institute of Public Health, Guangdong, China, 4 Yunnan Center for Disease Control and Prevention, Kunming, China, 5 Fred Hutchinson Cancer Research Center, Seattle, WA, United States Background: Dehong prefecture of Yunnan province was the gateway of China s AIDS epidemic. Most HIV- strains were first found in Dehong before spreading to other parts of the country. Study on HIV- genetic recombinants will provide key information on virus transmission dynamics and help to inform both local and national HIV prevention strategies. Methods: In Dehong, we surveyed all HIV infected young people (age 25 ) diagnosed in the first quarter of each year in Thepol genes fragment from a totalof 205 HIV- infected people were amplified and sequenced. The HIV- subtypes and recombinant patterns were identified by phylogenetic and breakpoint analyses. Near fulllength genome sequencing was performed to characterize the new recombinants and potential circulating recombinant forms (pcrfs).risk factors for generating various recombinant viral forms were analyzed by Chi-square tests and logistic regression methods. Results: Two thirds (3/205) of the HIV- infections in young people in Dehong were caused by new recombinant viruses. About 40% (54/3) of them were found to form transmission clusters, which was termed pcrfs. The rest (77/3) of them have not yet been able to form any transmission clusters and belonged to unique recombinant forms (URFs). The generation of the new HIV- recombinants was significantly associated with people with low education, residents outside the capital city of Dehong and being Myanmar residents. Conclusions: By properly sampling the young people, we revealed that the ongoing HIV epidemic in Dehong was caused by high proportion of new recombinant viruses, which was not found in sexually driven epidemic before. Considering Dehong had generated wide-spreading HIV- CRF_07, 08 BC, great efforts should be put on preventing the existing pcrf from further spreading and containing the URFs evolving to future CRFs. Prevention strategy should focus on those most at-risk for multiple exposures, residents with low education, on border region and migrating Burmese. 33

332 Posters Posters 39: Molecular Epidemiology P39. LB A New Cost-effective Viral RNA PCR-based Diagnostic for Detection of Early Subtype C HIV- Infection Debby Basu,2, Tyronza Sharkey 2, William Kilembe 2, Susan Allen 2,3, Eric Hunter Emory University, Emory Vaccine Center, Atlanta, GA, United States, 2 Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Emory University School of Public Health, Department of Global Health, Atlanta, GA, United States Background: Strategies for detecting early HIV- infection can be costly, inefficient and time-consuming, especially in settings where highthroughput screening is needed but viral load (VL) testing instruments may not be accessible or economical. We have developed a costeffective in-house viral RNA PCR that can empower limited-resource laboratories to screen for acute HIV- infection by amplification of viral regions in gag, pol and gp4. Methods: Viral RNA was extracted from plasma, and a multiplexed onestep reverse-transcriptase first round PCR, containing the outer primer sets for three regions in a single reaction, was performed. Nested second round PCR specific to each region followed. To validate that this multiplex assay could be used on batched plasma pools and to determine the limit of detection, we tested 4 samples of known VL, ranging from.2x0 7 to.x0 4 copies/ml, by diluting these plasmas from early infection :0 with uninfected plasma. Results: Positive amplification in at least of the 3 diagnostic regions was observed for all 4 mock-pooled plasma samples, with the majority (/4) showing positive amplification in all 3 regions. In the lowest VL case, input copies as low as an estimated 6 copies per first round PCR reaction still resulted in positive amplification in gp4. Conclusions: We have developed a multiplexed RT-PCR assay to detect early HIV- infection in patient plasma in a cost-effective, highthroughput manner. All samples tested were positive in at least one of three viral regions. The majority resulted in positive amplification in all three regions, thus showing potential universality of the assay in detecting viral RNA from Subtype C plasma samples with a range of VLs. This strategy costs less than $8/sample tested, and thus represents an economical strategy to screen patients for evidence of early HIV- infection in low-resource settings. This research was supported by NIH FIC R25 TW (DB), FIC 2D43 TW00042 (WK), RO MH (SA), R37 AI523and IAVI (SA). 332 HIV Research for Prevention 204 HIV R4P

333 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.0 Vaginal Concentrations of Lactic Acid Potently Inactivate HIV- Compared to Short Chain Fatty Acids Present During Bacterial Vaginosis Muriel Aldunate,2, David Tyssen, Catherine Latham, Paul Ramsland,2,3, Patrick Perlmutter 2, Thomas Moench 4, Richard Cone 5, Gilda Tachedjian,2,3 Burnet Institute, Centre for Biomedical Research, Melbourne, Australia, 2 Monash University, Microbiology Department, Melbourne, Australia, 3 University of Melbourne, Department of Microbiology and Immunology, Melbourne, Australia, 4 ReProtect Inc., Baltimore, MD, United States, 5 Johns Hopkins University, Department of Biophysics, Baltimore, MD, United States P40.02 Effects of Endogenous and Exogenous Female Reproductive Hormones on Gene Expression and Barrier Function in Female Genital Epithelia Ayesha Islam, Jai G. Marathe 2, Jeff Pudney 3, Seyoum Ayehunie 4, Robin R. Ingalls 5, Deborah J. Anderson 6 Boston University, OB/GYN, Boston, MA, United States, 2 Boston University, School of Medicine, Boston, MA, United States, 3 Boston University, Boston, MA, United States, 4 MatTek Corp, Ashland, MA, United States, 5 Boston Medical Center, Infectious Diseases, Boston, MA, United States, 6 Boston University, OB/GYN, Microbiology, Boston, MA, United States Background: Bacterial vaginosis (BV) is caused by an imbalance in vaginal microflora and is a major risk factor for sexually transmitted infections, including HIV in women. Microflora composition is likely influenced by low vaginal ph (~3.8), maintained by racemic DL-lactic acid (LA) (~0mM). BV alters the ph (>4.5) and the short chain fatty acid (SCFA) profile to predominantly acetic acid (BV) vs LA (non-bv). Our previous studies highlight the potent HIV virucidal activity of LA; however, the virucidal activity of BV-associated SCFAs is unknown. Methods: Virucidal activity of physiologically relevant non-bv associated SCFAs at ph 3.8 versus BV-associated SCFAs at ph 5 were compared against subtype B transmitted/founder (T/F) strains, RHPA and CH058, subtype C 92BR025 and subtype EA CMU02. Anti-HIV activity of 00mM of DL-LA (ph3.8) and 00mM of acetic acid (ph 5) was determined over time. The structure activity relationship (SAR) of SCFAs and HIV- virucidal activity was assessed at the same ph and at equal concentrations of the active protonated forms. Results: Non-BV associated SCFAs (ph3.8) rapidly inactivated T/F strains causing a 000-fold drop in HIV- infectivity while BV associated SCFAs (ph5) caused little inactivation. This potent virucidal activity could be attributed to DL-LA (a non-bv SCFA), and not low ph. DL-LA had the greatest virucidal activity against subtypes B, C and EA over related BVassociated SCFAs at the same ph and concentration of the active form. SCFA SAR analysis revealed potent virucidal activity is associated with the presence of hydroxyl groups, especially on the α-carbon; which is attenuated by the presence of a CH 3 group on the carboxylic acid. Conclusions: We show that LA, a non-bv SCFA, is a more potent HIV virucide than SCFAs produced during BV, suggesting that BV-associated SCFAs are not as protective for the female reproductive tract. SAR analysis reveals chemical elements required for HIV- activity that could inform the design of SCFA analogues. Background: Little is known about how female reproductive hormones estradiol-7β (E) and progesterone (P) influence vaginal barrier and immune function. Furthermore the synthetic progestin contraceptive Depo-Provera (DMPA) promotes vaginal SIV acquisition in macaques and may enhance HIV acquisition in women. We have studied the effects of endogenous and exogenous hormones on vaginal epithelial barrier function and molecular mechanisms of immune defense. Methods: We conducted an Affymetrix.0 ST microarray study to examine gene expression in MatTek vaginal (VEC) and endocervical (VEN) tissues after differentiation in media containing physiologic E (75nM) or E+P (75 and 700nM, respectively) or 30 nm DMPA for 0 days. To assess barrier function, tissues were seeded apically with CMFDA-stained macrophages and infiltration was assessed by confocal microscopy. Results: Our study confirms the hormonal responsiveness of these tissues, and identifies several genes that are significantly up and downregulated following exposure to hormones. Pathways identified by DAVID and Ingenuity Pathway Analysis (IPA) reflect classical hormone responses and epithelial differentiation, as well as a number of others that potentially affect HIV and other sexually transmitted infection acquisition including mediators of innate immunity, cell death, and tight junction molecules. VEC-DMPA showed increased membrane lipid storage but decreased steroid (E) responses and retinol metabolism. VEC-E showed increased lysozyme expression (3x) and decreased Caspse4 (-8x) expression versus hormone untreated VEC tissue (p< 0.05). Notably, gene expression profiles of VEN were distinct from VEC. E treatment of VEC prevented infiltration of macrophages by >50%, providing further evidence of its barrier enhancing effects. Conclusions: Female reproductive hormones and DMPA have distinct effects on molecular pathways underlying immune defense in vaginal and endocervical epithelium. E appears to fortify vaginal barrier function

334 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.03 P40.04 Softcup Compared to Cervicovaginal Lavage Sampling: Determining Total and HIVspecific IgGs in the Female Genital Tract - A Randomized Study Derseree Archary, Lenine Julie Liebenberg, Lise Werner, Sahil Tulsi, Nelisile Majola, Nivashnee Naicker, Sarah Dlamini, Natasha Samsunder, Salim S. Abdool-Karim,2, Jo-Ann S. Passmore,3,4, Lynn Morris 5, Nigel Garrett Centre for AIDS Programme of Research in South Africa (CAPRISA), Durban, South Africa, 2 Mailman School of Public Health, Columbia University, Department of Epidemiology, New York, NY, United States, 3 University of Cape Town, Cape Town, South Africa, 4 National Health Laboratory Services, Cape Town, South Africa, 5 National Institute for Communicable Diseases, Johannesburg, South Africa Background: Determining optimal methods for genital specimen collection for detection and quantification of mucosal immune responses is a priority for the HIV vaccine prevention field. Here we investigated whether Softcup (EuroFemPro, Netherlands) sampling yields greater quantities of HIV-specific immunoglobulins (IgG) and total IgG when compared to the standard, cervicovaginal lavage (CVL) method in a randomized study. Methods: Forty HIV-infected antiretroviral-naïve women from an HIV cohort study were randomized to undergo either Softcup sampling followed by CVL, or CVL alone. We measured HIV-specific IgGs against Gag p24, p66 (RT), gp4mn and gp20 in 9 Softcups, their matching CVLs and 20 randomized CVLs using Luminex multiplexing assays. Results: The average time of Softcup insertion was 4 minutes (range 75-43). Eighteen of 9 Softcup samples had HIV-specific IgG to all four antigens compared to 7/20 randomized CVLs. Detection of IgG to gp4mn was highest with gp20 the lowest for both Softcup and CVL. All four HIV-specific IgGs (MFIs) were significantly higher when Softcup was compared to randomized CVL (all p< 0.00). Higher specific activity [MFI/total IgG (ng/ml)] for Gag p24, gp4mn and gp20 (all p< 0.05) in Softcup compared to CVL was found, indicating that even lowly expressed gp20 IgG was significantly detected from Softcup. Recovery of total IgG from Softcup was significantly higher than from randomized CVLs (p< 0.000). Softcup collection did not compromise detectable HIV-specific IgGs in the subsequent CVL with no differences of the IgG titres of the matching CVL and randomized CVL samples; and total IgG in Softcup and matching CVL were highly correlated (r=0.73; p=0.0006). Conclusions: Softcup sampling is an ideal replacement to CVL alone, and a subsequent lavage after Softcup provides equivalent IgG detection. These data suggest that Softcup sampling should be considered a replacement method to assess antibody responses at the female genital tract. Proteomics Based Methods for Toxicity Monitoring of Rectal Microbicides Adam Burgener, Florian Hladik 2, Kenzie Birse 3, Ian McGowan 4 Public Health Agency of Canada, Winnipeg, MB, Canada, 2 University of Washington, Seattle, WA, United States, 3 University of Manitoba, Winnipeg, MB, Canada, 4 Microbicide Trials Network, Pittsburgh, PA, United States Background: The HIV prevention field requires better tools for studying mucosal drug toxicity and general inflammation. Standard tools that study individual components in isolation likely underestimates the global effects that may underpin complex immunological processes. Advances in mass spectrometry allow for the measurement of a large number of immunological parameters simultaneously. Here we evaluate the utility of this technology for examining rectal mucosal samples, and the effect of the known mucosal irritant nonoxynol-9 on the host rectal mucosal proteome. Methods: Rectal sponge samples were collected from 7 individuals pre (Visit ) and after 7 consecutive days (Visit 3) of once-daily doses of Nonoxynol-9 (N9) or placebo HEC gel controls (MTN-007 phase clinical trial). Sponge eluates were analyzed by tandem mass spectrometry. Results: 488 unique proteins were identified, that covered many functions that may be important for product toxicity including dermatitis (57, 6.78E-29), skin development (23, 4.57E-8), allergic response (47, p=3.0e-20), psoriasis (27, p= 5.67E-38), cell death (27, 3.68E- 6), necrosis (07, 4.82E-3), and cancer (279,.E-0). 46 proteins were differentially expressed (p< 0.05) between N9 and HEC controls, after 7 days of exposure (V3) (filtered to non-treated controls (n=8, p< 0.05). Hierarchical clustering showed a general overexpression of proteins in the N9 treatment arm. Biofunctional analysis indicated these factors belong to skin disorders (20) and general inflammation (), and apoptotic, carcinogenic, and pro-inflammatory canonical pathways, indicating a wide range of inflammatory and toxic effects of N9 (p< 0.05, BH corrected). Conclusions: This indicates that proteomics methods are suitable for monitoring immune factors in rectal mucosa, and mucosal-irritant associated responses. This shows the utility of systems biology techniques and adds to the available toolsets for toxicity monitoring of future microbicide candidates. 334 HIV Research for Prevention 204 HIV R4P

335 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.05 The Action of Cilia in the Upper Reproductive Tract of Women Influences the Distribution of HIV Ann M. Carias, Daniel Stieh, Danijela Maric, Thomas J. Hope Northwestern University, Chicago, IL, United States P40.06 Functional Assessment of Antibody Activity in Mucosal Tissue Explant and Cellular Inhibition Assays Hannah M. Cheeseman, Katja Klein, Abbey Evans, Deborah King, Robin J. Shattock Imperial College London, Group of Mucosal Immunology and Infection, London, United Kingdom Background: Understanding the precise mechanisms of HIV transmission across genital barriers is essential for studying vaccine development and inhibitory compounds. Previously, we illustrated that HIV can penetrate the intact female epithelium both ex vivo and in vivo, suggesting a diffusion mechanism for HIV entry; however, these studies were primarily focused on the lower female reproductive tract (FRT). We recently found that SIV based vectors can reach the ovary. The distance to transverse the uterus and fallopian tube of the upper FRT to reach the ovary is too far to reach by simple diffusion. Methods: To investigate the possible role of cilia on the diffusion of HIV in the upper FRT, we examined the localization of cilia by staining for microtubules. Bundles of microtubules are present in the extended structure of individual cilia. To assess how ciliated epithelium might influence the movement of fluorescent beads and virus, we directly visualized the dynamics of fluorescent particles using time-lapse imaging. Results: We find that the movement of cilia has a significant influence on the diffusion of virus and nanoparticles. First, the beating of the cilia accelerates and alters the diffusion of particles by causing convection currents in the regions adjacent to the ciliated epithelia. Second, the beating of the cilia pushes the particles away from the ciliated epithelium, essentially preventing particles from interacting directly with the underlying epithelial barrier. Conclusions: These data reveal that the ciliated epithelium of the upper FRT provides protection from potential contacting particles. It seems likely that this action also drives an acceleration of the diffusion of the particles. The net effect of the lack of binding to the tissue and acceleration drives particles to traverse the long distance to the ovary. These data suggest that the anatomy of the FRT, combined with the action of cilia, must be taken into account to achieve complete protection by inhibitory modalities. Background: The RV44 vaccine trial yielded results of 3.2% efficacy in protection from HIV- infection, despite a lack of broadly neutralising antibodies (BnAb). Subsequent analysis has demonstrated a correlate of protection was high ADCC (antibody-derived cellular cytotoxicity) IgG and low levels of IgA antibodies to the VV2 region of env. Here, we investigate the ability of a range of neutralising (nab) and nonneutralising (nnab) antibodies to prevent or lower HIV- infection in cellular and tissue explant models. Methods: A range of monoclonal antibodies with discrete epitope specificity and structure isolated by the CHAVI consortium from vaccinated and infected subjects have been screened in cellular and tissue based models designed to mimic the initial events in transmission. Results: A range of nnabs and nabs were screened for activity against HIV- BaL infection of macrophages, dendritic cells, dendritic trans/cis infection of T cells, and mucosal explants. 2/25 nnabs displayed no antiviral activity in these models. Four nnabs had inhibitory activity against macrophage infection, however none had any inhibitory activity against infection of mucosal tissue explants (rectal, penile, cervical). Only the nab CH3 displayed inhibitory activity across all models. Conclusions: To date only nab have been able to prevent infection across all models tested. These data suggest that nnabs may be insufficient to prevent the initial infection of target cells in mucosal tissue. Nevertheless, these conclusions come with caveats. First, experiments were performed with HIV- BaL, there may be differential activity against other strains. Second, polyclonal nnabs may show increased function. Third, the tissue explant models represent a static system with no influx of effector cells. Although such explants contain numerous cells capable of performing effector functions, it cannot be excluded that initial infection in vivo could result in recruitment of additional effector cells able to control or eliminate infection

336 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.07 P40.08 Genital IRIS, Immune Activation and Inflammation in the Female Genital Tract Influences HIV Shedding in HIV-infected Women Starting HAART Smritee Dabee, Jean-Mari Kriek, David Lewis 2,3, Maseko Venessa 2, Mkhize Nonhlanhla 2, Pamela P. Gumbi 3, Zizipho Mbulawa, Anna-Lise Williamson,4, Heather B. Jaspan,4, Jo-Ann S. Passmore,4 Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Department of Clinical Laboratory Sciences, Cape Town, South Africa, 2 National Institute for Communicable Diseases, Johannesburg, South Africa, 3 Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 4 National Health Laboratory Services, Cape Town, South Africa Background: Initiation of HAART has previously been associated with development of genital IRIS. As more HIV+ individuals are eligible to start treatment in sub-saharan Africa, reactivation of subclinical sexually transmitted infections (STIs) may influence genital HIV shedding and transmission to uninfected partners. We investigated the impact of HAART on STI prevalence, genital immune activation, inflammation, and genital HIV viral loads in HIV+ women initiating HAART. Methods: Blood and cervical mononuclear cells were obtained from HIV+ women before, and month, after starting HAART. T-cell activation and proliferation (CD38, HLA-DR, Ki67) were measured by FACS. IL- β, IL-6, IL-8, IP-0, MIP-α, MIP-β, TNF-α, IL-7, G-CSF and IL-0 concentrations were measured by Luminex. Screening for C. trachomatis, N. gonnorhoea, M. genitalium, T. vaginalis, and HPV was performed. Results: Although HAART significantly reduced blood T-cell activation (CD38 +, HLADR +, CD38 + HLADR + ), genital activation levels remained high. Cytokine concentrations were significantly higher in CVL than blood. Blood cytokine concentrations were generally lower after month on HAART, significantly so for IP-0 (p=0.05). In contrast, cytokine levels in CVL remained unchanged, with the exception of IL-0 which was lower after HAART (p=0.007). More than half of the HIV-infected women had a bacterial STIs and 90% were infected with HPV before starting HAART. Initiation of HAART did not change this prevalence. Whereas >85% of women had detectable HIV in their genital secretions pre-haart, 8% still had detectable genital tract viral loads month post-haart. Conclusions: On-going immune activation, inflammation and prevalent STIs in the female genital tract of women initiating HAART may influence risk for HIV transmission to uninfected partners, despite the beneficial effect of HAART to the individual. Effects of Intra-vaginal Drying Agents on Mucosal Immune Activation and Risk for HIV Acquisition in South African Women Kathleen E. Doherty, Melis Anahtar 2, Musie Ghebremichael 2, Christina Thogabekale 3, Nikita Padavattan 3, Krista Dong 4, Bruce D. Walker 2, Thumbi Ndung u 3,4, Douglas S. Kwon 2 Vanderbilt University, Nashville, TN, United States, 2 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 3 HIV Pathogenesis Programme, Durban, South Africa, 4 KwaZulu-Natal Research Institute for TB&HIV, Durban, South Africa Background: Most HIV transmissions occur in women following exposure to virus in the female genital tract (FGT). Because FGT inflammation has been associated with increased risk of HIV acquisition, it is critical to understand the biologic and behavioral factors that may contribute to elevated FGT inflammation. The Kwazulu-Natal (KZN) region in South Africa has one of the highest rates of intra-vaginal drying agent (IDA) usage worldwide. However, little is known about the immunologic effects of these agents, specifically as they relate to genital inflammation and risk of HIV acquisition. Methods: FGT samples and detailed behavioral data were collected from a high-risk cohort of young, HIV-negative women in KZN. Cytobrush cells were assessed by flow cytometry. FGT mononuclear cells and cervical tissue explants were incubated with drying agents in vitro and assessed for changes in immune activation. Results: Overall, 5.2% of the cohort reported using IDAs. The most commonly used agents were ntsu (powdered tobacco) and china fruit. Associations were seen between IDA use and earlier sexual debut (p=0.0305), older sexual partners (p=0.0073), frequency of sex (p=0.0373) and, importantly, increased HIV acquisition rates (p=0.005). IDA use was also associated with altered frequencies and activation states of immune populations at the cervicovaginal junction. Several common IDAs were shown to have effects on immune activation on FGT mucosal cells and cervical explants in vitro. Conclusions: This study describes the behavioral and immunological associations of IDA usage and their potential impact on HIV acquisition. It reveals increased acquisition rates among those who use IDA that are likely due to both behavioral and biological factors. This work lays the foundation for future research on therapies and behavioral interventions to modulate the immune landscape in the FGT to prevent HIV acquisition. 336 HIV Research for Prevention 204 HIV R4P

337 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.09 Acceptability of Using the Softcup for the Collection of Genital Fluid for Mucosal Assays in HIV Prevention Trials Nigel Garrett, Lindiwe Mpanza, Themba Cekwane, Nivashnee Naicker, Lise Werner, Nelisile Majola, Adolphus Mntambo, Derseree Archary Centre for the AIDS Programme of Research in South Africa (CAPRISA), Durban, South Africa P40.0 Alterations in Genital Tract Soluble Immune Mediators in HIV Positive Postmenopausal Women: Implications for HIV Acquisition and Transmission Josie Delisle, Ifeyinwa Benyeogor, Mariel Jais, Naji Younes, Mary Young 2, Mimi Ghosh The George Washington University, Washington, DC, United States, 2 Women s Interagency HIV Study, Washington, DC, United States Background: A simple, scalable and participant-friendly method for genital specimen collection to measure mucosal immune responses is needed for the HIV prevention field. The purpose of this study was to assess the acceptability of using Softcup, a menstrual cup, for the collection of genital fluid from women participating in an existing HIV cohort study. Methods: Forty HIV-infected antiretroviral-naïve women in a cohort study were randomized to have their genital sampling either by ) Softcup (EuroFemPro, Netherlands) followed by cervico-vaginal lavage (CVL), or 2) CVL alone, which is the current standard method for mucosal sampling. Softcups were clinician-inserted and were removed after 2 hours. Questionnaires with 5-point Likert scales ranging from extremely unacceptable/ uncomfortable to very acceptable/ comfortable were distributed to all women after removal of the Softcup, and were completed by the participants. Results: Twenty women, median age 3 (IQR 28-32), had a Softcup inserted. The average time from Softcup insertion to removal was 4 minutes (range 75-43). A total of 7/20 questionnaires were returned (85% response rate). Sixteen (94%) of participants answered that insertion and wearing the Softcup were very comfortable. All participants stated that the removal procedure was very comfortable, and that the Softcup procedure was very acceptable compared to their previous experience of genital examinations with a speculum performed by the clinician. All participants indicated their willingness to wear a Softcup again, and all preferred to have the Softcup inserted instead of a speculum. However, only 29% indicated that they would consider selfinserting the Softcup, if trained to do so. Conclusions: Collecting genital secretions from women using Softcup was highly acceptable and was preferred to speculum insertion used for other mucosal sampling methods. The Softcup should be considered for genital sampling in women in future HIV prevention trials. Background: Heterosexual transmission accounts for the majority of new HIV infections with women being more likely than men to be infected during vaginal intercourse. Multiple soluble immune mediators in the female reproductive tract (FRT) that are hormonally regulated are protective against HIV. It is unclear as to whether loss of estradiol in postmenopausal women results in a blunted innate immune response in the FRT thereby making them more susceptible to acquiring and transmitting HIV. Furthermore, it is unknown whether postmenopausal women on hormone replacement therapy (HRT) might recover these innate immune functions. In this study, we investigated changes in soluble immune mediators, IL-6, TNF-a, IL-8, MIP3a, Elafin, SLPI and Human beta defensin-2 (HBD-2) in cervical-vaginal lavage (CVL) of HIV(+) and HIV(-) postmenopausal women compared to premenopausal women and women on HRT. Methods: CVL from HIV(+) and HIV(-) premenopausal, postmenopausal and women on HRT were obtained from the Washington DC Women s Interagency HIV Study (WIHS) local repository and analyzed using commercially available ELISA. None of the HIV(+) women were on HAART. Statistical analyses were performed using the Kruskal-Wallis test (Graphpad Prism). Results: HIV(+) postmenopausal women had significantly higher plasma viral load and lower CD4 counts compared to premenopausal women. We also observed significantly lower levels of HBD2 and MIP3a in postmenopausal women, from both HIV(+) and HIV(-) groups. In HIV(-) postmenopausal women and women on HRT, significant decreases in levels of TNF-a, IL-6, and anti-hiv protective factor SLPI was observed. However, both TNF-a and IL-6 were increased in HIV(+) postmenopausal women. Conclusions: Our data indicate that both HIV status and menopausal status can dysregulate levels of soluble immune mediators in the FRT. A shift in the balance between proinflammatory factors and anti-hiv protective factors can result in higher susceptibility to acquisition and transmission of HIV in these women

338 Posters Posters 40: Mucosal Immune Activation and Inflammation P40. P40.2 Altered Levels of Soluble Immune Mediators in HIV-negative Postmenopausal Women: Implications for HIV Acquisition in the Elderly Mariel Jais, Naji Younes 2, Susan Cu-Uvin 3, Mimi Ghosh 2 Milken Institute School for Public Health, The George Washington University, Epidemiology and Biostatistics, Washington, DC, United States, 2 Milken Institute School fo Public Health, The George Washington University, Epidemiology and Biostatistics, Washington, DC, United States, 3 Brown University, Miriam Hospital, Providence, RI, United States Background: The female reproductive tract (FRT) secretes immune mediators protective against sexually transmitted infections, including HIV. As multiple immune factors in FRT are hormone-responsive, the loss of sex hormones with aging may undermine these defense mechanisms. Women are disproportionately affected by the HIV/AIDS epidemic with heterosexual contact being the major source of new infections. Reports indicate older women are sexually active and often do not use protection as pregnancy is a non-issue. Therefore, investigating the effects of sex hormone-loss on FRT mucosal immune factors is an important target to curtail HIV acquisition. Methods: CVL samples were collected from 20 HIV-negative premenopausal and postmenopausal women. Each postmenopausal woman provided one sample; each premenopausal woman provided 3 samples collected during proliferative, ovulatory, and secretory stages of menstrual cycle. Commercially available ELISA kits were used to assess the levels of IL-6, IL-8, TNFa, Elafin, HBD-2, MIP3a/CCL20 and SLPI. Samples were analyzed for their anti-hiv activity against HIV- IIIB and BaL strains via the TZM-bl assay. Results: We observed significantly lower levels of critical immune mediators in CVL from postmenopausal women compared to those from premenopausal women: TNFa (.6 vs 5.5 pg/ml), MIP3a (.0 vs 93.8 pg/ml), SLPI (39.6 vs pg/ml) and HBD-2 (626 vs 682 pg/ml). Levels of IL-6 and IL-8 displayed a trend toward lower levels in postmenopausal samples whereas Elafin levels remained unchanged. Inhibition of HIV- infection was observed for X4/IIIB and R5/BaL strains in both pre and postmenopausal samples with inhibition of BaL stronger in premenopausal samples (54.2 vs 37.6%). Conclusions: Our findings indicate that levels of critical mucosal immune factors and anti-hiv- activity in CVLs are affected by the hormonal status of healthy HIV-negative women. This indicates the need for specific therapeutic interventions to boost genital tract immunity against HIV in older women. Bacterial Vaginosis and HIV: An Analysis of the MDP30 Trial Sarah Joseph, Nathlee Abbai 2, Sinead Delaney-Moretlwe 3, Mitzy Gafos, Saidi Kapiga 4, Maureen Chisembele 5, Andrew Abaasa 6, Ute Jensch 7, Suzanna Francis 8, Sheena McCormack, Richard Hayes 8, Angela Crook MRC Clinical Trials Unit at UCl, HIV Prevention, London, United Kingdom, 2 Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 3 Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 4 Mwanza Interventions Trials Unit, Mwanza, Tanzania, United Republic of, 5 University Teaching Hospital, Obstetrics and Gynaecology, Lusaka, Zambia, 6 MRC/UVRI, Uganda Research Unit on AIDS, Entebbe, Uganda, 7 Clinical Laboratory Services, University of Witwatersrand, School of Pathology, Johannesburg, South Africa, 8 London School of Hygiene & Tropical Medicine, MRC Tropical Epidemiology Group, London, United Kingdom Background: Bacterial vaginosis (BV) has been shown to be associated with increased susceptibility to HIV- infection. We have used data from the MDP30 microbicide trial to estimate the effect of BV on the risk of HIV acquisition. Methods: 849 HIV negative women, enrolled at 6 centres in 4 sub- Saharan African countries between 2005 and 2008, and with a BV result at baseline, were included in the analysis. BV was assessed using the Ison Hay method: (negative, reference), 2 (intermediate) or 3 (positive) at weeks 0, 2, 24, 40 and 52. Cox proportional hazard models were fitted to estimate the effect of BV on risk of HIV acquisition. Data were censored at HIV sero-conversion or at the last visit attended. The following baseline or time-updated covariates were considered in a multivariate model: age, centre, gel group, condom use, sexual frequency, abnormal vaginal discharge, HSV-2, other cervico-vaginal infection (CT, NG, TV, syphilis) and reported contraception type. Results: 384 women had HIV seroconverted by the end of follow-up (8002 person-years). At baseline, 350 (37%) women were positive for BV, 2073 (24%) were intermediate and 3268 (38%) negative. 3066/849 (36%) never tested positive for BV whereas 82 (2%) tested positive throughout. Adjusted hazard ratios (ahr) for the risk of HIV were.45 (95% CI ) for those testing positive for BV and ahr.04 (95% CI ) for those with an intermediate result. Other covariates which remained independently associated with HIV acquisition in the adjusted model were age, centre, abnormal vaginal discharge, HSV-2, other cervico-vaginal infection and use of injectable DMPA contraception. Conclusions: The results of this secondary analysis show positive BV to be independently associated with increased risk of HIV acquisition, further strengthening the case for more research into this potentially neglected risk factor for HIV. 338 HIV Research for Prevention 204 HIV R4P

339 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.3 P40.4 Dynamic and Divergent Bacterial Species Similar to Bacterial Vaginosis Prior to SIV in Pigtail Macaques Nichole R. Klatt, Sujatha Srinivasan 2, Laura Richert-Spuhler, Michael Koday, David Fredricks 2 University of Washington, Pharmaceutics, WaNPRC, Seattle, WA, United States, 2 Fred Hutchinson Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, United States Background: A challenge in developing interventions to prevent mucosal HIV transmission is incomplete understanding of correlates of vaginal transmission, including mucosal inflammation. Vaginal SIV/SHIV transmission in pigtail macaques is an excellent model for HIV transmission, however little is known about how genital bacterial species may influence inflammation and SIV transmission. Methods: Broad-range 6S rrna gene PCR and pyrosequencing was performed on vaginal swabs for bacterial identification using a cohort of pigtail macaques in a cross-sectional analysis (N=22), and a more limited longitudinal analysis (N=3). Bacteria were identified to the species or genus level using a custom designed reference set of sequences, providing comprehensive characterization compared to previous studies focused on genera and phylum. Results: Pigtail macaques had diverse microbial communities at all phases of the menstrual cycle, with a range of dominant bacteria in different animals, including Prevotella, Porphyromonas, Dialister, Clostridiales, Bacteroides, Streptococcus and Lactobacillus species. In contrast to previous studies, we found three animals had a Lactobacillusdominant vaginal microbiota, all at peak sex skin swelling (indicating ovulation). Several species were identified that are commonly found in bacterial vaginosis (BV) in humans, including Atopobium vaginae, Prevotella buccalis, BVAB2, Peptoniphilus lacrimalis, Prevotella timonensis and Gardnerella vaginalis. Longitudinal sampling demonstrated that vaginal bacterial communities were dynamic and possibly driven by alterations in environment throughout the menstrual cycle. Conclusions: The macaque vaginal microbiota is diverse, dynamic, and can resemble the human vaginal microbiota, including BV-associated bacteria as well as Lactobacillus spp. These data provide a foundation for understanding how vaginal microbial communities may impact risk of HIV/SIV transmission using these models. Impact of Persistent Human Papillomavirus (HPV) Infections on Inflammatory Cytokine Levels in the Female Genital Tract: Implications for HIV Risk Jean-Mari Kriek, Shameem Z. Jaumdally, Zizipho Mbulawa,2, Pamela P. Gumbi, Lindi Masson, Shaun L. Barnabas,3, David Coetzee 2, Anna-Lise Williamson,2, Francesca Little 4, Jo-Ann S. Passmore,2 Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 2 National Health Laboratory Service, Cape Town, South Africa, 3 Desmond Tutu HIV Foundation, IIDMM University of Cape Town, Cape Town, South Africa, 4 University of Cape Town, Department of Statistical Science, Cape Town, South Africa Background: Women with persistent HPV-infections are at increased risk for developing cervical cancer. Clearance of HPV-infections has been associated with genital inflammation. HPV- infection has been shown as a risk factor for HIV acquisition along with genital inflammation. The aim of this study was to evaluate the impact of genital tract inflammation and HPV persistence or clearance in HIV negative women. Methods: Cervical samples were collected from 38 HIV-negative women at two time points, six months apart. IL-8, IL-6, IL-0, IL-5, IL-2p40, IP- 0, MCP-, MIP-a, MIP-b, IL-a, IL-b, eotaxin, fractalkine, and G-CSF concentrations were measured by Luminex at enrollment. HPV types were assessed at both time points using the Roche Linear Array HPV Genotyping assay. Results: There were 20/38 HPV-infected women at enrollment [9/38 had high-risk (HR) and /38 had low-risk (LR) HPV types]. An additional 7 women acquired an HPV infection over the 6 months of follow-up. Of the 20 women initially infected with HPV, only 2/20 cleared their infections while 8/20 had infections that persisted for 6 months. Women with HR HPV at enrollment generally had higher cytokine concentrations in their genital tracts than women with LR types, although none of the cytokine evaluated were significantly different between groups. Women acquiring an HPV infection over 6 months had lower overall genital inflammation at enrolment compared to women who remained HPVnegative (p=0.005). In contrast, women with persistent HPV infections generally had increased inflammation at enrolment compared to women who remained negative (p=0.049). Conclusions: Although HPV infection has been associated with increased risk for HIV infection, we found that HPV infections generally did not cause an inflammatory response in the female genital tract

340 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.5 Genital Tract Immunological Markers in Sub- Saharan African Women with Relevance to HIV Risk and Prevention P40.6 Influence of Vaginal Microbiota on the Diffusional Barrier Properties of Cervicovaginal Mucus Jordan K. Kyongo, Tania Crucitti 2, Joris Menten 2, Liselotte Hardy 2,3, Piet Cools 4, Johan Michiels, Sinead Delany-Moretlwe 5, Mary Mwaura 6, Gilles Ndayisaba 7, Sarah Joseph 8, Raina Fichorova 9, Janneke van de Wijgert 0, Guido Vanham,,2, Kevin K. Ariën, Vicky Jespers 3, Biomarkers Study Group Institute of Tropical Medicine, Department of Biomedical Sciences, Antwerp, Belgium, 2 Institute of Tropical Medicine, Department of Clinical Sciences, Antwerp, Belgium, 3 Institute of Tropical Medicine, Department of Public Health, Antwerp, Belgium, 4 Ghent University, Department of Microbiology, Immunology and Clinical Chemistry, Ghent, Belgium, 5 University of the Witwatersrand, Wits Reproductive Health & HIV Institute, School of Clinical Medicine, Johannesburg, South Africa, 6 International Center for Reproductive Health, Mombasa, Kenya, 7 Project Ubuzima, Kigali, Rwanda, 8 MRC Clinical Trials Unit at University College London, London, United Kingdom, 9 Brigham and Women s Hospital, Harvard Medical School, Department of Obstetrics, Gynaecology and Reproductive Biology, Boston, MA, United States, 0 University of Liverpool, Institute of Infection and Global Health, Liverpool, United Kingdom, University of Antwerp, Faculty of Pharmaceutical, Veterinary and Biomedical Sciences, Antwerp, Belgium, 2 University of Brussels, Faculty of Medicine and Pharmacology, Brussels, Belgium Background: Data on immune mediators in the genital tract and the behavioural and clinical factors that modulate them in Sub-Saharan women are limited. Methods: Cervicovaginal lavage (CVL) samples from 430 sexually active women from Kenya, South Africa and Rwanda were analysed for twelve soluble immune mediators. qpcr was used to quantify ten bacterial species in vaginal swab samples. We also compared the anti-hiv activity of CVL samples from bacterial vaginosis (BV)-positive women to those from women with a Nugent score of 0. Results: Pregnant women, adolescents, women engaging in traditional vaginal practices and HIV-positive women differed in specific soluble markers compared to reference groups of adult HIV-negative women. An increase in cervical mucus, the presence of cervical ectopy, abnormal vaginal discharge and having multiple sexual partners were each associated with an increase in mediators of inflammation. Interleukin (IL)-α, IL-β, IL-6, IL-2 and IL-8 were elevated and the IL-RA/(IL-(α+β) ratio decreased in the CVLs of women with BV. Interferon gammainduced protein (IP)-0 was decreased in BV-positive compared to BVnegative women. Lactobacillus crispatus and Lactobacillus vaginalis were associated with lower levels of pro-inflammatory cytokines and each BV-associated species with increased pro-inflammatory cytokines. The in vitro anti-hiv activity of CVLs from BV-positive women was stronger than that of BV-negative women. Conclusions: We found significant associations of factors that can influence HIV susceptibility with the levels of soluble immune mediators in the vaginal environment of sexually active women. These factors need to be considered when establishing normative levels or pathogenic cut-offs of biomarkers of inflammation and associated risks in African women. IP-0 suppression may be one potential mechanism of immune evasion by BV-associated bacteria. Lastly, cervicovaginal secretions of women with BV may contain active anti-hiv substances that should be examined more closely. Kenetta Nunn, Ying-Ying Wang 2, Dimple Harit, Richard Cone 2, Samuel Lai University of North Carolina at Chapel Hill, Eshelman School of Pharmacy, Chapel Hill, NC, United States, 2 Johns Hopkins University, Baltimore, MD, United States Background: To reach target cells underlying the vaginal epithelium, HIV must penetrate cervicovaginal mucus (CVM) secretions. We previously found that native CVM from women with healthy, lactobacillidominated vaginal flora (ph ~3.5-4) effectively trapped HIV, but the same finding was not observed in a later follow-up study. We sought to investigate whether the vaginal microbiota can differentially influence the diffusional barrier properties of CVM against HIV and other sexually transmitted viruses. Methods: We evaluated the mobility of fluorescent HIV pseudoviruses in fresh, undiluted CVM from different women subjects using high resolution multiple particle tracking. We then correlated the observed mobility to various biochemical and biophysical characterizations, including ph, D- and L- lactic acid levels and Nugent scores. We are also performing microbiome analysis on a subset of the specimens. Results: In a cohort of over 40 CVM specimens, the real-time mobility of mcherry-gag labeled HIV was not significantly correlated to native ph, Nugent scores, total IgG/IgA or total lactic acid (LA) levels. Interestingly, CVM samples that failed to trap HIV exhibited substantially lower D-LA content, whereas CVM samples that effectively trapped HIV exhibited both high D- and L- LA. The same phenomenon was observed with ΔEnv virions, suggesting it may be universal among all enveloped viruses. Addition of D-LA did not trap HIV, suggesting that low D- lactic acid is likely a surrogate marker of the native CVM barrier. High L-LA/low D-LA is consistent with vaginal microbiota that is populated with specific strains of lactobacilli, which we are currently confirming using 6S rrna pyrosequencing. Conclusions: The vaginal microbiota plays an underappreciated and yet to be clarified role in modulating the innate diffusional barrier properties of mucus against HIV. Our work may help identify women who are at markedly enhanced susceptibility to acquiring HIV and other sexually transmitted infections. 340 HIV Research for Prevention 204 HIV R4P

341 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.7 P40.8 A Randomized Study Comparing Softcup and Cervicovaginal Lavage Sampling to Measure Genital Cytokine Concentrations in HIVinfected Women Lenine J. Liebenberg, Nigel Garrett, Lise Werner, Nelisile Majola, Nivashnee Naicker, Natasha Samsunder, Sarah Dlamini, Jo-Ann S. Passmore,2,3, Salim S. Abdool Karim,4, Derseree Archary CAPRISA, Durban, South Africa, 2 Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 3 National Health Laboratory Services, Cape Town, South Africa, 4 Department of Epidemiology, Mailman School of Public Health, Columbia University, New York, NY, United States Background: Evaluating concentrations of genital cytokines are key to understanding local immunity to HIV infections or infection risk. Menstrual softcups may provide a more convenient method for collection of genital specimens than cervicovaginal lavages (CVLs). We compared softcup sampling to the CVL collection method for detection of cytokines in genital fluid. Methods: Forty ART-naïve HIV-infected women from a cohort study of HIV infection were randomized to either have genital fluid collected by softcup with subsequent CVL, or by CVL alone. Luminex was used to measure the concentrations of 48 cytokines involved in hematopoiesis, regulation, adaptive responses, inflammation and growth, in both CVL and softcup specimens. Results: In randomized softcup specimens, 42/48 cytokines were consistently detected in all participants compared to 26/48 in CVL (p=0.0003). Cytokines detected in softcups but not in CVL included those important to HIV infection and pathogenesis (IL-8, MIP-α, RANTES, TNF-α). Differential concentrations were observed for 4/48 cytokines in randomized softcup and CVL specimens, and 22 of these cytokines remained significantly elevated in softcups after adjustment. Cytokine concentrations in matched softcup secretions and CVL correlated significantly (r=0.903; p< 0.000), suggesting that proportions of cytokines measured are retained in the two sampling methods. However, even in paired specimens, 88% of cytokines were consistently detected in softcup specimens from all participants compared to 69% in CVL (p=0.0263). Conclusions: This study demonstrates that the overall cytokine composition in softcup secretions compares to CVL genital sampling. However, softcup sampling allows for more reliable detection of cytokines, particularly those present at the lowest concentrations. Safety Studies of a Formulation Comprising Recombinant Human Surfactant Protein D for Prevention of HIV- Infection Taruna Madan, Hrishikesh Pandit, Kavita Kale, Uday Kishore 2 National Institute for Research in Reproductive Health (ICMR), Innate Immunity, Mumbai, India, 2 Brunel University, Centre for Infection, Immunity and Disease Mechanisms, London, United Kingdom Background: Owing to the lack of an effective vaccine for HIV-, efforts towards development of potent preventive strategies has gained impetus. We have previously reported that human surfactant protein D, an integral innate immune molecule has a potent anti-hiv- activity. SP-D interferes with CD4 binding regions of gp20 and modulates pro-inflammatory signaling cascade and cytokine production. Thus, in view of its potential as a microbicidal agent, we determined in vitro anti-hiv activity of a formulation comprising recombinant human SP-D (rhsp-d) against several HIV- clinical isolates, with different tropism and subtypes in various cell lines and primary cells. The formulation did not show any adverse effects on the viability or lactic acid/peroxide production of clinical strains of lactobacilli. Methods: In vitro post-coital efficacy of rhsp-d in the presence of vaginal lavage (VL) and seminal plasma (SP) was determined. Effect on viability of VK2 cells was determined by MTT assay. In the rabbit vaginal irritation model, ml gel containing rhsp-d formulation, placebo and 0.% SDS formulation as positive control was administered for 0 consecutive days. Vaginal tissues were examined by H&E staining for infiltration of immune cells and inflammation and the serum cytokine levels were determined. Results: The rhsp-d inhibited HIV- infectivity in a dose-dependent manner in the presence of VL and SP. We also observed that SP-D is expressed by vaginal epithelial cells and is hormonally regulated. rhsp-d had no adverse effect on the viability of VK2 cells with a therapeutic index >0. The formulation is being evaluated using rabbit vaginal irritation model and the results are being analyzed. Conclusions: The study suggests that rhsp-d formulation is promising with its efficacy and safety. Studies using human vaginal explants models would be the next step towards its translational potential. 34

342 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.9 Mucosal Proteomic Profiles Associated with Female Genital Tract Inflammation Kelly Arnold, Adam Burgener 2, Kenzie Birse 2, Laura Dunphy, Kamnoosh Shahabi 3, Max Abou 4, Jessie Kwatampora 5, Billy Nyanga 5, Joshua Kimani 2, Lenine Liebenberg 6, Lindi Masson 7, Salim S. Abdool Karim 6, Jo-Ann S. Passmore 7, Douglas A. Lauffenburger, Rupert Kaul 3, Lyle R. McKinnon 6 MIT, Boston, MA, United States, 2 University of Manitoba, Winnipeg, MB, Canada, 3 University of Toronto, Toronto, ON, Canada, 4 Public Health Agency of Canada, Winnipeg, MB, Canada, 5 University of Nairobi, Nairobi, Kenya, 6 CAPRISA, Durban, South Africa, 7 University of Cape Town, Cape Town, South Africa P40.20 Control of HIV- Infection in the Female Reproductive Tract by Mucosal Innate Immunity Determinants Héloïse Quillay,2, Hicham El Costa 2, Claude Cannou 2, Marion Duriez 2, Romain Marlin 3, Claire de Truchis 4, Anne Le Breton 4, Mona Rahmati 5, Julien Ighil 5, Olivier Schwartz 2, Françoise Barré- Sinoussi 2, Marie-Thérèse Nugeyre 2, Elisabeth Menu 2 Univ. Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France, 2 Institut Pasteur, Paris, France, 3 Université Bordeaux, Bordeaux, France, 4 Antoine Béclère Hospital, Clamart, France, 5 Pitié Salpétrière Hospital, Paris, France Background: Inflammatory cytokines in cervicovaginal lavage (CVL) were associated with an increased risk of HIV infection in the CAPRISA004 trial. The mechanisms by which inflammation increases HIV risk are not yet fully understood. We investigated mucosal biomarkers and pathways associated with raised inflammatory cytokines in the genital tract. Methods: We classified HIV-negative Kenyan women as inflammation+ (n=28) or inflammation- (n=68) based on the elevation (upper quartile) of at least three of 7 inflammatory cytokines MIP-3α, RANTES, IL-8, MIP- β, IL-β, IL-α, and GM-CSF in CVL. CD4+ T cells from endocervical cytobrushes were enumerated in the same participants. CVL samples were further analyzed by tandem mass spectrometry, and subjected to several canonical and biofunctional pathway analyses. Partial least squares discriminant analysis (PLSDA) and LASSO methods for regression shrinkage/selection were used to model the minimum set of biomarkers that best classified inflammation groups. Results: Women with inflammation had a >2 fold increase in the number of endocervical CD4+ T cells (p< 0.00). A total of 76 proteins were quantified; 53 of these were increased and 27 were decreased in inflammation+ women (p< 0.0). Of these 80 biomarkers, 50 remained significantly different after applying a 5% False Discovery Rate threshold. Hierarchical clustering suggested significantly different proteomes associated with mucosal inflammation (p< 0.0). Inflammation was associated with acute phase signaling, complement activation, and remodeling of epithelial adherens junctions, and the chemotaxis and adhesion of immune cells. Anti-inflammatory peptidase inhibitors and keratinization factors were downregulated. LASSO and PLSDA identified 3 biomarkers that distinguished the inflammation+ group with 88% accuracy on calibration and 84% accuracy on cross-validation. Conclusions: These analyses highlight several potential underlying molecular mechanisms that could link inflammation to HIV susceptibility. Background: Mucosas are the preferential portal for HIV- entry in the body. It is thus crucial to identify the determinants necessary for an efficient control of transmission in the mucosa. The control of HIV- infection at the materno-fetal interface during the first trimester of pregnancy is a good model to study natural protection against transmission in the female reproductive tract. In the decidua (uterine mucosa during pregnancy), macrophages (dms) are the main target cells of R5 tropic HIV-. The aim of this study was to characterize control mechanisms of infection in the decidua. Methods: Deciduas were obtained from HIV- negative women undergoing elective abortions (8-2 weeks of amenorrhea) with their written informed consent. Purified dms were infected with R5 HIV- or HIV-/VSV-G pseudotype. Natural killer (dnk) cells were added to dms (ratio :5) at different times. Cocultures were performed in the same well or separately in a double chamber system. SAMHD expression was determined by flow cytometry. In some experiments, VLP-Vpx was added to dms simultaneously with the virus. Results: Cocultures of infected dms with autologous dnk cells revealed that dnk cells controlled dm HIV- infection in the early steps of infection. Cell-to-cell contacts and soluble factors were necessary for an efficient control. NKG2D and IFN-γ were involved in the control. dms highly expressed SAMHD, a restriction factor blocking HIV- replication. Infection of dms in presence of the viral accessory protein Vpx, which degrades SAMHD, increased dm infection. Conclusions: These data demonstrate in vitro for the first time that ) dnk cells control HIV- infection by several mechanisms and 2) the infection of dms is restricted by SAMHD. The determinants of protection identified in the decidua are under investigation in the non pregnant female reproductive tract. These studies give important information for the development of future preventive strategies against HIV- mucosal transmission. 342 HIV Research for Prevention 204 HIV R4P

343 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.2 Presence of Male Partner Semen Influences the Inflammatory and Innate Cytokine Environment in the Female Genital Tract Sinaye Ngcapu, Tracey Meiring 2, Lindi Masson 2, Lise Werner, Lenine Liebenberg, Nigel Garrett, Koleka Mlisana,3, Carolyn Williamson 2, Quarraisha Abdool Karim, Salim Abdool Karim, Jo- Ann Passmore,2,4 CAPRISA, Durban, South Africa, 2 Institute of Infectious Diseases and Molecular Medicine, University of Cape Town, Cape Town, South Africa, 3 School of Laboratory Medicine and Medical Sciences, University of KwaZulu Natal, Durban, South Africa, 4 National Health Laboratory Services, Cape Town, South Africa P40.22 Acceptability of Multiple Mucosal Specimen Collection in a Phase HIV Vaccine Trial in Rwanda Julien M. Nyombayire, Rosine Ingabire, Jeannine Mukamuyango, Etienne Karita, Dagna Laufer 2, Harriet Park 2, Phillip Bergin 3, Frances Priddy 2, Susan Allen,4 Rwanda Zambia HIV Research Group-Projet San Francisco, Kigali, Rwanda, 2 International AIDS Vaccine Initiative, New York, NY, United States, 3 IAVI Human Immunology Laboratory, London, United Kingdom, 4 Emory University, School of Medicine, Department of Pathology and Laboratory Medicine, Atlanta, GA, United States Background: In addition to spermatozoa, semen contains high concentrations of anti-inflammatory cytokines (TGF-b, IL-0, PGE2), inflammatory cytokines (IL-8, IL-b, IL-6), and activated immune cells, each potentially capable of influencing the immune environment of the lower female genital tract. While several studies have quantified the inflammatory cytokine composition of genital secretions from sexually active women that may influence HIV risk, few have investigated the influence of recent sexual intercourse on the cytokine milieu of cervicovaginal secretions. Methods: We investigated the semen exposure in cervicovaginal lavage (CVL) samples collected from 72 high-risk HIV negative women by investigating the presence of Y-chromosome by amplification of the TSPY gene of the Y-chromosome by PCR, and by quantifying prostate specific antigen (PSA) levels by ELISA. Levels of 42 cervicovaginal cytokines, 5 MMPs and 4 TIMPs were measured by luminex and were compared in women with or without semen exposure. Results: Y-chromosomes were detected in 29% (2/72) of CVLs and denoted sex within 7 days prior to sampling (Y-chromo+); and7% (2/72) of participants were PSA+, denoting sex within 2 days prior to sampling. Of the Y-chromo+ participants, 57% (2/2) were also PSA+. Only 33% (7/2) of women who were Y-chromo+ and PSA+ had self-reported condom use at their last sex act. Y-chromo+ CVLs had significantly higher levels of PDGF-AA, MMP-7 and MMP-0 levels, after adjusting for multiple comparisons. In contrast, PSA+ CVLs had significantly lower concentrations of IL-Ra, IL-2Rα, GM-CSF and TNF-β. Conclusions: These findings suggest that the presence of semen influenced the cytokine and MMP profile in CVL and should be taken into consideration when investigating biological markers in the female genital tract. Background: HIV infection is mostly acquired through mucosal surfaces and an efficacious HIV vaccine would ideally elicit an immune response at these surfaces.this study presents acceptability of the mucosal sampling procedures among study volunteers enrolled in a phase HIV vaccine trial in Kigali, Rwanda. Methods: Since January 203, Project San Francisco (PSF) has been participating in a multicenter phase clinical trial to assess the safety and immunogenicity of a Sendai HIV vaccine given intra-nasally and an Ad35-GRIN HIV vaccine administered intra-muscularly in primeboost regimens.the study was discussed extensively with volunteers during education and consenting sessions.mucosal sampling included saliva from parotid glands (Salimetrics Oral Swabs), oral fluids(falcon tubes),nasal secretions(floq swabs),cervico-vaginal secretions (Softcup or Merocel sponge), and rectal secretions (Merocel sponge). Samples were collected at baseline before vaccination and at 5 subsequent time points during follow-up to date.the study is ongoing and will have 9 mucosal sampling time points in total.consent for mucosal sampling was reassessed at each collection visit. Results: A total of 20 participants were enrolled (8 women and 2men), and all 20 participants (00%) consented to each type of mucosal secretion collection.as of April 204, the 20 study volunteers have completed the 6 mucosal collection visits each. All study participants provided all protocol required specimens, both at baseline and at the 5 subsequent follow-up visits, resulting in a 00% acceptability rate. Conclusions: This study confirms that multiple collection sites and repeated mucosal specimen collection in HIV vaccine trials is highly acceptable in Rwanda. The high acceptability of these procedures may be a reflection of extensive counseling and mutual trust between study participants and study staff

344 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.23 P40.24 Implementation of a Training Program to Standardize Mucosal Sample Collection and Processing at Multiple Laboratories in Eastern Africa Gloria Omosa-Manyonyi, Robert Langat, Elizabeth Mutisya, Harriet Park 2, Philip Bergin 3, Bashir Farah, Hilda Ogutu, Simon Ogola, Gina Ouattara, Jackton Indangasi, Rose Ndambuki, Pat Fast 2, Dagna Laufer 2, Fran Priddy 2, Omu Anzala Kenya AIDS Vaccine Initiative Institute of Clinical Research, Nairobi, Kenya, 2 International AIDS Vaccine Initiative (IAVI), New York, NY, United States, 3 International AIDS Vaccine Initiative (IAVI)-HIL, Imperial College, London, United Kingdom Background: HIV- transmission occurs predominantly across the genital mucosa during sexual intercourse. Understanding host mucosal responses to HIV- infection, and assessing protective mucosal responses during microbicide and HIV vaccine clinical trials are important. For comparable data across studies and research centers, mucosal samples should be obtained in a standardized reproducible manner. Methods: KAVI-ICR and IAVI developed standardized protocols for collecting, processing and assay of cervico-vaginal, rectal and upper respiratory tract mucosal samples. After proof of concept studies, KAVI- ICR developed a 3-day-hands-on training program on specific mucosal sample collection and processing methods, and transferred these to research staff at centers across Eastern Africa. Participant consenting, community engagement, and access to supplies were discussed. Results: Standardized mucosal sample collection and processing procedures were applied to three Phase studies at KAVI-ICR; mucosal anti-hiv IgG/IgA responses were detectable. Forty-six research staff from 5 centers had hands-on training on the standardized methods (2 clinicians, 4 nurses, 20 lab staff). Projet San Francisco (PSF) Kigali Rwanda, and MRC/UVRI Uganda Research Unit on AIDS had on-site training; while staff from UVRI-IAVI HIV Program Uganda, KEMRI/ Walter Reed Project (KEMRI/WRP)-Kericho Kenya, and KEMRI-Centre of Geographical Medicine Research Coast Kenya, visited KAVI-ICR for training. PSF Rwanda and KEMRI/WRP-Kericho have applied the methods learnt on mucosal studies, including mucosal sampling in HIV vaccine trials. The mucosal studies at PSF Rwanda were multi-site and the resulting mucosal data was comparable across all participating sites. Conclusions: It is possible to standardize mucosal sampling across research centers to ensure comparable samples and resulting data. KAVI-ICR may become a regional mucosal training center for transfer of these methods in South-South interactions. Anti-HIV Activity of Vaginal Epithelial Cells and Vaginal Secretions Mickey V. Patel, Mimi Ghosh 2, Richard M. Rossoll, John V. Fahey, Charles R. Wira Geisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United States, 2 George Washington University, Epidemiology and Biostatistics, Washington, DC, United States Background: HIV is the leading cause of death for reproductiveage women, with the majority of transmission events occurring via heterosexual intercourse. The first site of exposure is the vagina, and understanding its immune environment, which changes with hormone levels across the menstrual cycle, is crucial to developing effective vaccines and prevention measures. Methods: Using a menstrual cup, we developed a novel technique to concurrently recover fresh undiluted vaginal secretions (VS) and vaginal epithelial cells (VEC) at mid-proliferative, ovulatory and mid-secretory stages of the menstrual cycle. VEC were isolated and treated in 96- well plates with estradiol (E 2 : 5x0-8 M), progesterone (P 4 : x0-7 M), or a combination of both for 48hrs after which conditioned media (CM) were recovered. VEC CM and VS were analyzed for presence of anti-hiv proteins by ELISA, and anti-hiv activity using a TZM-bl assay. Results: CCL20, RANTES, elafin, HBD2, SDF-α and IL-8 were present in VS. VS demonstrated significant inhibition of X4 (IIIB) HIV and increased infectivity of reporter cells by R5 (CH077.t) HIV. No inhibitory effect was seen for BaL and CH058. No significant differences in either antiviral protein concentration or anti-hiv activity with respect to menstrual cycle stage were measured, but marked differences were observed in both parameters over the course of the cycle between different women, and in consecutive cycles from the same woman. For VEC, E 2 significantly decreased the secretion of HBD2 and elafin over 48hrs. CM from E 2 - or P 4 -treated VEC had no anti-hiv activity. However, CM from E 2 /P 4 cotreated cells significantly inhibited both R5 and X4 HIV. Conclusions: There are multiple levels of protection in the vagina. The sensitivity of VEC to E 2 and P 4 suggests that their contribution to immune protection varies across the menstrual cycle. The variation in anti-hiv activity in VS demonstrates that immune protection is not constant and that factors in addition to hormones influence antiviral protection. 344 HIV Research for Prevention 204 HIV R4P

345 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.25 P40.26 Antiviral Responses of Fibroblasts in the Female Reproductive Tract Mickey V. Patel, John V. Fahey, Richard M. Rossoll, Charles R. Wira Geisel School of Medicine at Dartmouth, Physiology and Neurobiology, Lebanon, NH, United States Background: The female reproductive tract (FRT) is the primary location of heterosexual transmission of HIV. However, the intricacies of immune protection at this site are not well understood, in particular the contribution of stromal fibroblasts in preventing HIV infection. Fibroblasts are the predominant cell type in the sub-epithelial layers of the FRT, and the initial stages of HIV infection occur in their presence. Methods: Benign FRT tissues from the endometrium (EM), endocervix (Cx) and ectocervix (ECx), were recovered from HIV- women undergoing hysterectomy. Following enzymatic digestion, cells were passed through nylon filters to isolate purified populations of fibroblasts. These were maintained in culture for 4-6 days prior to treatment with estradiol (E 2 : 5x0-8 M) for up to 72hrs in the presence or absence of viral stimuli. Conditioned media (CM) was recovered and analyzed for the presence of anti-hiv proteins (ELISA) and anti-hiv activity (TZM-bl). Results: The anti-hiv proteins RANTES, SDF-α and CCL20 were present in CM. While average RANTES levels were relatively constant between the 3 tissue sites, SDF-α was significantly higher in CM from EM fibroblasts (2526 pg/ml) compared to the Cx (276 pg/ml) and ECx (962 pg/ml). In contrast CCL20 was highest in CM from the Cx (500 pg/ ml) compared to the EM (40 pg/ml) and ECx (7 pg/ml). E 2, via ERα, significantly upregulated the secretion of SDF-α by only EM fibroblasts, with no effect on RANTES and CCL20. CM from all 3 sites showed potent inhibition of BaL (R5 HIV). However, CM from E 2 -treated EM fibroblasts demonstrated significantly greater inhibition of IIIB (X4 HIV) compared to control CM. In contrast, control ECx CM significantly upregulated IIIB infection of TZM-bl cells. Conclusions: These results demonstrate that secretions from stromal fibroblasts are capable of inhibiting HIV infection. Furthermore, their antiviral activity is altered by E 2 suggesting that it may vary across the menstrual cycle and by anatomical location in the FRT. Soluble Toll-like Receptor 2 Is Significantly Elevated in HIV- Infected Breast Milk and Inhibits HIV-induced Cellular Activation and Infection Bethany M. Henrick,2, Xiao-Dan Yao,2, Anna G. Drannik,2, Alash le Abimiku 3, Kenneth L. Rosenthal,2, INFANT Study Team McMaster University, Department of Pathology & Molecular Medicine, Hamilton, ON, Canada, 2 McMaster University, McMaster Immunology Research Centre, Hamilton, ON, Canada, 3 Institute of Human Virology, University of Maryland School of Medicine, Baltimore, MD, United States Background: We previously demonstrated that immunodepletion of soluble Toll-like receptor 2 (stlr2) from breast milk significantly increased HIV infection in vitro; therefore we next wanted to identify its mechanism of action and characterize stlr2 levels in HIV-infected and uninfected breast milk. Methods: Breast milk from HIV-infected and uninfected Nigerian and Canadian women were blindly tested for levels of stlr2, proinflammatory cytokines, and viral antigenemia. In vitro experiments were conducted using cell lines and primary cells to assess stlr2 function on innate responses and HIV infection. Results: HIV-infected breast milk showed significantly increased levels of stlr2 compared to uninfected milk. Further, stlr2 levels correlated with HIV p24 and IL-5, thus suggesting a local innate compensatory response in the HIV-infected breast. Given the increase in stlr2 in HIV-infected milk, we next demonstrated that mammary epithelial cells (MECs) and macrophages, which are prevalent in milk, produced significantly increased levels of stlr2 following exposure to HIV- proteins p7, p24 and gp4 or the TLR2 ligand, Pam3CSK 4. stlr2 coimmunoprecipitated with p7, p24 and gp4 and inhibited HIV-induced NFκB activation and inflammation. Importantly, binding of stlr2 to HIV proteins inhibited a TLR2-dependent increase in CCR5 expression, thus resulting in significantly reduced HIV infection. Conclusions: Our results demonstrate that stlr2 inhibits HIV cellular activation, inflammation, increased CCR5 expression and HIV infection through direct interaction with the virus. Further, stlr2 is significantly elevated in HIV-infected breast milk and positively correlated with p24 and IL-5. Overall, our data suggests that stlr2 may play a critical role in inhibiting mother-to-child HIV transmission

346 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.27 An Approach to Unravel Cellular Mechanisms Responsible for Enhanced Neisseria gonorrhea Induced HIV Acquisition and its Effect on Microbicides P40.28 Unexpected Inflammatory Effects of Intravaginal Gels (Universal Placebo Gel and Nonoxynol-9) on the Upper Female Reproductive Tract Anwesha Sanyal, Phalguni Gupta, Ming Ding, Deena Ratner University of Pittsburgh Graduate School of Public Health, Infectious Diseases and Microbiology, Pittsburgh, PA, United States Background: Sexually Transmitted Infection, like Neisseria gonorrhea (NG) in the female reproductive tract (FRT) increases sexual transmission of HIV-. Our goal is to identify NG induced cellular and viral factors that are responsible for enhanced HIV acquisition. Methods: Cervical tissues in an organ culture system were used to measure inflammatory response to NG by real time RT-PCR and MSD multiplex assay, respectively. HIV transcription in the presence of NG was measured in a reporter gene assay in TZM bl cells. Microdisected epithelial layer from cervical tissues following exposure to NG and HIV were subjected to comprehensive transcriptome analysis using second generation sequencing in an ion torrent platform. Effect of NG on microbicides activity was evaluated in an organ culture system. Results: Upon exposure to NG high levels of inflammatory cytokines IL-β and TNFα were detected in cervical tissues. Furthermore, the supernatants from the organ cultures exposed to NG increased transcription from the HIV- LTR in a TZM bl cell based assay and increased transmission of HIV across cervical mucosa. From gene expression analysis of the microdisected epithelial layer of the tissues exposed to NG and the control, 30 genes have been identified to be differentially upregulated with high statistical significance. CXCL0 and IL8 genes were found to be common between tissue exposed to HIV and NG group. In addition, the efficacy of microbicide (RC0 and CSIC), which decreased HIV transmission in organ culture, was not compromised in the presence of NG in the organ culture system. Conclusions: The results suggest that NG infection induces higher levels of CXCL0 and IL8 proteins that either by themselves or interact with cellular factors responsible for HIV transmission and enhance HIV acquisition. Furthermore, the presence of NG did not have any effect on the antiviral activity of the microbicides indicating that these microbicides would be effective for decreasing HIV transmission in the presence of existing NG. Karen Smith-McCune, Joseph Chen, Ruth Greenblatt, Barbara Shacklett 2, Uma Shanmugasundaram 2, Joan Hilton, Brittni Johnson,3, Fatima Barragan,4, Juan Irwin, Margaret Takeda, Jane Pannell, Jean Perry, Linda Giudice UCSF, San Francisco, CA, United States, 2 UC Davis, Davis, CA, United States, 3 UCLA, Los Angeles, CA, United States, 4 Michigan State University, Grand Rapids, MI, United States Background: Intravaginal microbicides could provide women with a selfcontrolled means for HIV prevention, but results from clinical trials have been largely disappointing. We postulated that unrecognized effects of intravaginal gels on the upper female reproductive tract (FRT) might contribute to the lower-than-expected efficacy of HIV microbicides. Methods: In this observational crossover study, 28 healthy female volunteers used no product (control cycle) or nightly application of either intravaginal nonoxynol-9 gel [N9] (an agent with known harmful effects on the lower FRT) or the universal placebo gel [UPG] (an agent used as a placebo control in many microbicide trials) from the end of menses to the mid-luteal phase (intervention cycles). They then underwent sample collection for measurements of T-cell phenotypes, transcriptional profiling, and protein concentrations from 3 anatomic sites above the vagina: the cervical transformation zone (TZ), the endocervix and the endometrium. We used hierarchical statistical models to estimate mean (95% CI) intervention:control fold-changes in relevant phenotype levels. Results: Exposure to N9 and UPG generated a common harm signal that included transcriptional up-regulation of inflammatory genes CCL20 and IL8 in the cervical TZ, decreased protein concentrations of secretory leukocyte protease inhibitor in endocervical fluid, increased percentages of terminally differentiated CD4+ effector T-cells in the endocervix, and transcriptional up-regulation of inflammatory mediators KIR3DS, glycodelin-a, and osteopontin in the endometrium. Conclusions: These results underscore the need to consider the effects of gel vehicles as well as microbicide agents on the upper FRT in studies of vaginal microbicides. Given the pro-inflammatory effects of UPG on the upper FRT, it may not be a suitable placebo for microbicide trials. Supported by NIH grants #AI (PI: Warner Greene), #U54HD (to L.C.G.), and #F32HD (to J.C.C.) 346 HIV Research for Prevention 204 HIV R4P

347 Posters 40: Mucosal Immune Activation and Inflammation Thursday, 30 October P40.29 Lactic Acid, a Vaginal Microbiota Metabolite, Elicits an Anti-inflammatory Response from Vaginal and Cervical Epithelial Cells Anna Hearps,2, Raffi Gugasyan, Daniela Srbinovski,2, David Tyssen, Muriel Aldunate,2, Deborah J. Anderson 3, Richard Cone 4, Gilda Tachedjian,2,5 Burnet Institute, Centre for Biomedical Research, Melbourne, Australia, 2 Monash University, Melbourne, Australia, 3 Boston University, Boston, MA, United States, 4 Johns Hopkins University, Baltimore, MD, United States, 5 The University of Melbourne, Department of Microbiology and Immunology, Melbourne, Australia P40.30 Gene Expression Analysis of Human Vaginal Mucosal Response to Pro-inflammatory Stimuli - Identification of Biomarkers of Vaginal Inflammation Irina A. Zalenskaya, Andrea R. Thurman, Neelima Chandra, Nazita Yousefieh, Suzanne S. Jackson, Sharon Anderson, Gustavo F. Doncel CONRAD, Eastern Virginia Medical School, OB/GYN, Norfolk, VA, United States Background: Lactobacilli sp. dominate the vaginal microbiota in about /3 of reproductive age women and HIV susceptibility increases with a shift from lactobacilli to bacterial vaginosis associated bacteria. Lactobacilli acidify the vagina to ph< 4.0 by producing ~% lactic acid (LA) in a nearly racemic mixture of D and L isomers. We have shown that 0.3% L-LA has potent HIV virucidal activity. Here we investigate if LA elicits an anti-inflammatory response on epithelial cells from the lower female reproductive tract (FRT) that might play a role in decreasing HIV susceptibility. Methods: The effect of LA in the apical medium was assessed on vaginal (VK2), endocervical (End) and ectocervical (Ect) epithelial cells grown in transwells. Toxicity effects were determined by viability staining and diffusion of fluorescently labelled dextrans through the cell layer. Cytokine profile from epithelial cells was determined following stimulation with toll-like receptor (TLR) agonists ± LA. Results: L-LA (0.3%; ph 3.9) had little impact on VK2, End and Ect monolayer toxicity. Stimulation of all epithelial cell types with poly(i:c) (TLR3) induced high-levels of pro-inflammatory cytokines IL-6 and IL- 8, and their variable induction with TLR agonists Pam(3)CSK(4)(TLR/2) and lipopolysaccharide (TLR4). In contrast, the presence of 0.3% L-LA or D-LA either significantly abrogated or reduced TLR-induced IL-6 and IL-8 secretion by epithelial cells. Irrespective of the TLR agonists, L-LA and D-LA elicited high-levels of the anti-inflammatory cytokine IL-RA (~30,000 pg/ml) from all cell types. Neither 0.3% L-LA at neutral ph nor acidity alone (HCl) increased IL-RA or decreased TLR induced IL-6 and IL-8 production. Conclusions: LA at ph 3.9 found in lactobacillus-dominated vaginal microbiota elicits an anti-inflammatory effect on lower FRT epithelial cells and inhibits inflammation induced by bacterial and viral TLR agonists suggesting a role in mitigating inflammation-induced HIV susceptibility at the genital mucosa. Background: Understanding the mechanisms underlying mucosal susceptibility to HIV infection and defining new biomarkers of vaginal inflammation and immune activation are essential to developing safe and effective HIV prevention products. To this end we analyzed transcriptomes of human vaginal tissues after in vivo treatment with proinflammatory (PIC) and non-inflammatory (NIC) compounds. Methods: Nineteen women (mean age= 32) applied 2-4 doses of PICs, including imiquimod and nonoxynol-9 (N9), and NIC, hydroxyethyl cellulose (HEC)-based placebo gel, intravaginally, in a crossover design. Vaginal biopsies were taken at baseline and after treatment and preserved in RNAlater or formalin. Gene expression (n = 93 tissue samples) was analyzed by using Affymetrix U33 Plus 2 arrays. Data processing was done using BRB-array tools software. Genes showing statistically different expression (p< 0.00) between treatment and control groups and fold change differences 2 were considered differentially expressed. Functional analysis was done using Ingenuity Pathway Analysis and published data. Immunohistochemical analysis (IHC) was performed on paraffin embedded vaginal tissues. Results: Transcriptomic analysis indicated that HEC did not alter mucosal gene expression significantly. Conversely, imiquimod caused dysregulation of 879 genes, and N9 of 89 genes. There were 66 genes common to both PIC treatments. Functional analysis indicated 27 genes involved in inflammatory and immune responses, 8 of which were chemoattractants for immune cells. Upstream regulation analysis revealed strong interferon signature. IHC showed PIC-induced influx of immune cells, including CD3 + T cells, into the vaginal mucosa. Conclusions: Intravaginal application of PICs induced a strong mucosal immuno-inflammatory response and transcriptomic change with a common profile of dysregulated genes which can serve as biomarkers of vaginal inflammation and immune activation in the safety evaluation of HIV prevention interventions

348 Posters Posters 40: Mucosal Immune Activation and Inflammation P40.3 Cervical Epithelial Cells from HIV--Exposed Seronegative Sex Workers Express a Distinct Cytokine/Chemokine Profile upon Toll-like Receptor Activation Nyla Dil, Joshua Kimani 2, Makobu Kimani 2, Frank Plummer 3, Blake T. Ball University of Manitoba/Public Health Agency of Canada, Medical Microbiology and Infectious Diseases/National Lab for HIV Immunology, Winnipeg, MB, Canada, 2 University of Nairobi, Medical Microbiology, Nairobi, Kenya, 3 University of Manitoba/National Microbiology Lab, Winnipeg, MB, Canada Background: Majority of human HIV- infections are acquired through heterosexual transmission across female genital mucosa. Epithelial cells that line the genital mucosa express toll-like receptors (TLR) and act as first line of defense against mucosal infections. Immune locale orchestrated by TLR mediated activation of female genital epithelial cells can be a critical determinant of HIV- resistance or susceptibility. In this study we investigated the role of TLR signaling in female genital epithelial cells in determining the local mucosal cytokine /chemokines milieu. Methods: Endoervical cytobrsuh samples were obtained from Pumwani CSWs cohort in Nairobi, Kenya. HESN (n=22), HIV- (n=24) and HIV+ (n=23).cervical epithelial cells (CECs) were purified through a series of nylon membrane filtrations. Purity and viability of CECs was assessed by Ber-EP4 expression and MTS assay, respectively. CECs were cultured with or without TLR3 ligand (poly:ic) or TLR-9 ligand combined for 24 h. Cytokine/chemokines levels in the supernatants were determined using the Milliplex MAP multiplex kit (Human Cytokine/Chemokine I and III) and analyzed on the BioPlex-200 according to manufacturer s protocol. Results: We tested 28 cytokines and chemokines (GM-CSF, IFNa2, IFNgamma,IL-a, IL-b, IL-2, IL-6, IL-7, IL-8, IL-0, IL-2 (p40), IL-2(p70), IL-5, IL-7, IP-0, MCP-, MCP-3, MDC, MIP-a, MIP-b, RANTES, Fractalkine, GRO, TNFa, I-TAC, MIG, MIP-3 a, and MIP-3 b). CECs from HESN individuals expressed significantly lower levels of interferon-γinduced protein 0 (IP-0), IL-8, IL- a, MIG, and MIP-3 a upon TLR3 or combined TLR-9 stimulation compared with CECs from HIV- and HIV+ women. These cytokines and chemokines have important functions in inflammatory and immune cell recruitment. Conclusions: These results highlight the role of TLR signaling in CECs and support the immune quiescence model of HIV- protection, whereby lower target and inflammatory cell recruitment at the genital mucosa reduces HIV- target cell numbers in HESN. 348 HIV Research for Prevention 204 HIV R4P

349 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.0 Immunogenicity of Recombinant Semliki Forest Virus-based RNA Vaccines Expressing HIV- Indian Subtype C Antigens in Mice Seema Ajbani, Shilpa Velhal, Ravindra Kadam, Vainav Patel, Atmaram Bandivdekar National Institute for Research in Reproductive Health (ICMR), Department of Biochemistry and Virology, Mumbai, India P4.02 Antiviral Activity of 5-Hydroxytyrosol, a Microbicidal Candidate against HIV- Transmission Jose Alcami, Luis-Miguel Bedoya, Patricia Obregón, Manuela Beltran, Eduardo Gomez-Acebo 2, David Auñón 2, Laura Capa Instituto de Salud Carlos III, AIDS Immunopathogenesis Unit, Majadahonda, Spain, 2 Seprox Biotech SL, Madrid, Spain Background: Genetic vaccinations have generally involved immunization with antigen-encoding nucleic acid, usually DNA. RNA vaccination based on engineered RNA replicons derived from several RNA viruses is gaining increased attention and several vaccines are currently being investigated for infectious diseases, cancer and allergies. Present study investigates the potential of recombinant Semliki Forest virus replicon RNA expressing HIV- antigens from Indian subtype C isolates to generate cellular and humoral immune responses in mice. Methods: Balb/C mice were immunized with recombinant Semliki Forest virus (SFV2gen) RNA encoding HIV-C gag or env antigen from Indian primary isolates. Three intramuscular injections of rsfv2gen/gag RNA or rsfv2gen/env RNA were given at 2 week intervals in the hind legs. Splenocytes were harvested at necropsy and analyzed for antigenspecific T cell responses by IFN-γ ELISPOT assay and Intracellular cytokine staining (ICCS) assay using antigen-specific peptide pools representing HIV-C Gag/Env consensus sequences. Sera of mice injected with rsfv2gen/env RNA were evaluated for HIV-C Env-specific binding antibodies by gp20 ELISA. Results: Gag-specific IFN-γ T cell responses were detected in all animals vaccinated with rsfv2gen/gag RNA following three immunizations. Surprisingly although Env-specific IFN-γ positive T cells were below the detection limit of ELISPOT assay, an assessment of vaccine-induced IL-2 secretion by intracellular staining followed by flow cytometry revealed Env-specific IL-2 production which was mediated by both CD4+ and CD8+ T cells. Serological analysis revealed HIV-C Env-specific binding antibody responses in mice receiving rsfv2gen/env RNA. Conclusions: These findings demonstrate that SFV replicon-based recombinant RNA vaccines were capable of eliciting HIV-C antigenspecific cellular and humoral immune responses in mice. These constructs are currently being investigated for their immunogenicity as a component of a multi-gene vaccine. Background: We are currently developing an effective and low-cost microbicide based on 5-Hydroxytyrosol (5-HT), a molecule not used as microbicide previously that displays both anti-inflammatory and antiviral properties. The objective of this work is to analyze the activity against HIV- of 5-HT, alone or in combination with other antiviral molecules in different cellular systems and its toxicty in vitro and in vivo. Methods: The antiviral activity of 5-HT was assessed against SIV and different HIV- strains (B and C clades) using different experimental models: cell lines, lymphocytes and monocytes from peripheral blood and autologous co-culture of DC-SIGN expressing cells and lymphocytes. Anti-HIV activity of 5-HT was assessed alone or in combination with Tenofovir. Synergism was analyzed according to T-C Chou and P. Talalay method (Trends Pharmacol. Sci. 4, ). Local tolerability at vaginal mucosa of 5-HT was assessed in rabbits (n=6) at two different concentrations (90 and 397mg/L) during 7 consecutive days by topical route. Results: 5-HT was active against SIV and different HIV- clades with IC50 ranging between 20-30µM. 5-HT was also active in the context of the immune synapse with IC50 in the 5µM range. In vitro toxicity was not observed at doses of.000 µm. Topical administration of 5-HT did not cause irritative responses or morphological alteration in the vaginal mucosa of rabbit. A strong synergistic effect was displayed by combination of 5-HT and Tenofovir (ED50 Combination index = 0.237). Conclusions:. 5-HT was active against SIV and different HIV- strains in vitro in 2. 5-HT inhibited HIV infection of lymphocytes, monocytes and was active in the context of the immune synapse, 3. Strong synergistic activity with Tenofovir was found, 4. No toxicity was observed in vitro and in vivo at the doses tested. In summary 5-hydroxytyrosol is a new class of microbicide combining both anti-inflammatory and anti-hiv properties and represents a potential candidate for future clinical trials

350 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.03 P4.04 Co-delivery of Antigen p24 and NOD-ligands by PLA Nanoparticles to Human Dendritic Cells Promote Highly Functional HIV-- Specific T-cell Responses Núria Climent, Vincent Pavot 2, Felipe García, Thierry Lioux 3, Eric Perouzel 3, Charlotte Primard 2, Stéphane Paul 4, Jose María Gatell, Montserrat Plana, Bernard Verrier 2, Teresa Gallart CELLEX-IDIBAPS-HIVACAT, Barcelona, Spain, 2 Institut de Biologie et Chimie des Protéines, UMR 5305 CNRS/UCBL, Cedex 07, Lyon, France, 3 CAYLA, InvivoGen, Toulouse, France, 4 Groupe Immunité des Muqueuses et Agents Pathogènes, INSERM CIE 3 Vaccinologie, Faculté de Médecine, Saint-Etienne, France Background: A recently explored approach for HIV vaccination is the administration of a variety of antigens and adjuvants included in nanoparticles targeted to dendritic cells (DC) to potentiate antigenspecific T-cell responses and a protective immunity without having detrimental effects associated to viral vector. The use of TLR ligands as mucosal adjuvant for vaccine administration has already been largely studied, but the use of NOD-like receptor (NLR) ligands needs to be further investigated. This study tested polylactic acid (PLA) colloidal biodegradable nanoparticles, coated with HIV Gag (p24) and encapsulating Nod ligands or 2 (NodL and NodL2) as adjuvant. Methods: We performed different ex vivo assays with monocyte derived DC (MDDC) including capture of the different nanoparticle formulations and induction of maturation. Co-cultures between MDDC and autologous lymphocytes were performed in order to measure proliferation by CFSE (FACS) and cytokine secretion (25-plex luminex) from asymptomatic HIV + -infected patients (n=). Results: PLA-p24-NodL and PLA-p24-NodL2 nanoparticles were highly captured by MDDC from HIV- individuals and induced a high degree of MDDC maturation compared with p24 or PLA-p24 (p< 0.05). In accordance, PLA-NodL and PLA-p24-NodL2 nanoparticles increased HIV-specific CD4 + and CD8 + T-cell proliferation being PLA-p24-NodL2 more than 6-folds higher compared with p24 or PLA-p24 (p< 0.0). These compounds induced highly functional cytokine secretion (IFN-γ, IL-2R, IL-6 and MIP-β/CCL4) in the autologous co-culture compared with p24 or PLA-p24 (p< 0.05). In all the cases, the highest immunoresponse was induced by PLA-p24-NodL2 (p< 0.0). Conclusions: Co-delivery of antigen p24 and NOD ligands by PLA nanoparticles to dendritic cells enhances antigen capture, DC-maturation and improves highly functional HIV-specific cellular immune responses from HIV+ patients. These compounds would be useful as novel therapeutic and preventive approaches in the context of HIV- vaccine. Vaccine Antigen Design to Maximize anti-hiv CD4+ T-cell Responses: From Mice to Nonhuman Primates Edecio Cunha-Neto, Susan Ribeiro, Daniela Santoro-Rosa 2, Rafael Almeida, Vinicius Santana, Jorge Kalil University of São Paulo, São Paulo, Brazil, 2 Federal University of São Paulo, São Paulo, Brazil Background: Purposely eliciting CD4+ T cell responses has been essentially unexplored in the HIV vaccine field, despite increasing evidence for the importance of the CD4+ T cell response in protection against HIV. Methods: We used rational vaccine design to develop a DNA vaccine encoding HIV- B subtype or M-type conserved, multiple HLA-DRbinding CD4+ T cell epitopes, found to be recognized by multiple HIV- -infected patients. Results: Vaccines elicited broad, polyfunctional, and long-lived CD4 + T cell responses in BALB/c and several HLA class II transgenic mice, eliciting extensive cross-clade immunity. Immunization of BALB/c mice increased CD8+ T cell responses against subsequent whole HIV protein immunization, and reduced vital titers after challenge with a recombinant vaccinia virus encoding HIV proteins. Immunization prior to recombinant gp40 HIV envelope protein drastically increased the IgG2a/IgG ratio of elicited anti-gp40 antibodies. Immunization of Rhesus macaques under electroporation induced broad IFN-γ ELISPOT responses 0-fold higher than those found in mice; we found a predominantly CD4+ T cell intracellular cytokine response basically consisting of IFNγ, TNFα IL-2, Granzyme B. Conclusions: By virtue of inducing broad responses against multiple conserved CD4+ T cell epitopes that can be recognized across widely diverse, common HLA class II alleles, this vaccine concept may induce T cell responses against multiple peptides in large proportion of the genetically heterogeneous population. By increasing the chance of matching the responses with multiple epitopes in the infecting HIV isolate, the vaccine concept may also cope with HIV genetic variability. The vaccine concept may be a candidate for standalone use or in association with conventional immunogens, to increase the amplitude, coverage and effectiveness of the induced response. Pending new results with immunization of Rhesus macaques with different viral vectors and prime-boosting with gp40, a Phase I clinical trial will be launched. 350 HIV Research for Prevention 204 HIV R4P

351 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.05 P4.06 Vaccinia Virus with Selective Deletions Enhances T Cell Response to HIV Antigens by Specific Neutrophil Recruitment Mauro Di Pilato, Ernesto Mejías-Pérez, Manuela Zonca 2, Beatriz Perdiguero, Carmen Elena Gómez, Marianna Trakala 3, Jacobo Nieto, José Luis Nájera, Carlos Oscar S. Sorzano 4, Lourdes Planelles 2, Mariano Esteban Spanish National Centre for Biotechnology (CNB), Department of Molecular and Cellular Biology, Madrid, Spain, 2 Spanish National Centre for Biotechnology (CNB), Department of Immunology and Oncology, Madrid, Spain, 3 Spanish National Cancer Research Centre (CNIO), Cell Division and Cancer Group, Madrid, Spain, 4 Spanish National Centre for Biotechnology (CNB), Biocomputing Unit, Madrid, Spain Background: In the context of vaccinia virus (VACV) infection, it was previously known that neutrophil infiltration is tissue-protective and that neutrophils generate virus-specific memory CD8 T cells, transporting antigens from the dermis to the bone marrow. However, it was not known how neutrophil recruitment is modulated by VACV, factors involved and significance in triggering immune responses. Methods: Here we describe the generation of an attenuated VACV strain (NYVAC) that expresses HIV- clade C antigens but lacks three specific viral genes (A52R, K7R and B5R). We analyze the capacity of NYVAC-C ΔA52RΔB5RΔK7R to activate NFκB signaling pathway, to induce specific innate immune response, and to generate adaptive immune response to HIV- antigens compared to NYVAC-C. Results: A52R, K7R and B5R cooperate to inhibit the NFκB signaling pathway, and the triple ablation in modified virus restores NFκB function in macrophages. After NYVAC-C ΔA52RΔB5RΔK7R virus infection, NFκB pathway activation leads to expression of several cytokines and chemokines that recruit specific neutrophil populations (Nα and Nβ) to the infection site. Neutrophils display features of antigen-presenting cells, and their trafficking to the infection site and to various lymphoid organs is essential to enhance the T-cell response to HIV Gag and Pol antigens. Conclusions: Based on these inherent properties, the NYVAC-C ΔA52RΔB5RΔK7R represents an effective vector vaccine candidate for prophylactic and therapeutic settings, and provides a basis for the design of new vaccines. Endosymbiont Trichomonas Vaginalis Virus as a Target for HIV Prevention Titilayo Fashemi, Hidemi Yamamoto, Nibert Max 2, Athe Tsibris,2, Raina Fichorova,2 Brigham and Women s Hospital, Boston, MA, United States, 2 Harvard Medical School, Boston, MA, United States Background: Trichomoniasis caused by the genitourinary protozoan parasite Trichomonas vaginalis (TV) is one of the leading risk factors of genital HIV shedding, sexual and perinatal HIV transmission. The majority of TV clinical isolates carry endosymbiont dsrna viruses (TVV). We have recently shown that TVV causes a profound TLR-3- dependent inflammatory reaction by human cervical and vaginal epithelial cells; however, the effects of TVV on HIV host cells and HIV replication have not been investigated to date. We hypothesized that the immunoinflammatory responses to TVV are at least partially responsible for trichomoniasis-attributable HIV risk. Methods: Peripheral blood mononuclear cells were stimulated with clinical TV isolates and purified TVV, followed by infection with primary isolates of CXCR4- and CCR5-tropic HIV-. Supernatants were collected at multiple time points over a period of 5 days to measure HIV- p24 by ELISA and immune mediators by multiplex immunoassays. ANOVA was applied, and P< 0.05 was considered significant. Results: TVV-positive TV induced significantly higher proinflammatory responses as compared to TVV negative TV isolates. TVV-positive TV up-regulated proinflammatory and Th/Th2 cytokines and enhanced those responses when applied together with other immune stimulants e.g. IL-2 and PHA. While levels of proinflammatory responses (IL--beta, TNF-alpha and IL-8) showed a tendency to subside after 72h, the levels of immunoregulatory cytokines e.g. IFN-gamma, IL-4, IL-5, IL-3 and IL-2p70 continued to rise. Purified TVV virions induced comparable immune activation. TVV+TV pre-exposure caused a 50-fold increase in p24 levels and the enhancement lasted for at least 5 days (p< 0.00). Conclusions: Our data support the hypothesis that TVV virus causes inflammation and immune cell activation capable of enhancing and propagating HIV replication and thus should be further investigated as a plausible target for HIV prophylaxis. (NIAID HU CFAR 5P30AI , R56AI09889 and R0AI079085). 35

352 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.07 P4.08 Deletion of Immunomodulatory A44L, A46R and C2L Viral Genes from Modified Vaccinia Ankara (MVA) Genome: Effect on its Immunogenicity Maria Pia Holgado, Cynthia Maeto, Juliana Falivene, Yanina Ghiglione, Maria Paula del Medico Zajac 2, Gabriela Calamante 2, Maria Magdalena Gherardi Instituto de Investigaciones Biomedicas en Retrovirus y SIDA, Buenos Aires, Argentina, 2 Instituto de Biotecnologia, CICVyA - INTA, Castelar, Argentina Background: MVA still retains genes involved in host immune response evasion. We have previously reported the optimization of its vaccine potential after removing the C2L gene, coding an IL8 binding protein. Here we analyze the immunogenicity of MVA vectors harboring the simultaneous deletion of two viral genes: A44L, implicated in synthesis of steroid hormones and A46R, which inhibits transduction signals from TLR (MVAΔA44L-A46R: MVAd); or including C2L deletion also (MVAΔC2L/ΔA44L-A46R: MVAt) Methods: C57Bl/6 mice were immunized with MVAwt or deleted MVAs (ΔMVAs). We evaluated the adaptive T-cell response to VV (Vaccinia Virus) epitopes at acute (7 dpi) and memory phases (45 dpi) in spleen and draining lymph nodes (DLNs). The amount of IFNγ and IL2 producing cells was measured by ELISPOT. We studied the percentage of specific cytotoxic CD8 T-cells by flow cytometry, and the response of memory T-cells among specific proliferating CD8 T-cells. To study the innate response, we immunized mice with MVAwt or MVAt and pattern of cytokines produced were evaluated between 0-24 hpi Results: At 7 dpi both ΔMVAs induced significant increases in IFNγ anti-vv CD8 and CD4-T cells vs MVAwt (.5 to two-fold,p< 0.0;2.5 to five-fold, p< 0.0 respectively), and IL2 anti-vv CD8 T- cells (up to five-fold higher;p< 0.0). Notably, ΔMVAs still elicited a higher response than MVAwt at 45 dpi (p< 0.05). Proliferating anti-vv CD8 T-cells were augmented from % (MVAwt) to 3% (MVAt). Moreover, this vector elicited a higher proportion of specific TCM compared to MVAwt (45% vs 20%). The percentage of specific cytotoxic CD8 T-cells secreting IFNγ was also improved by MVAt. The innate response analysis showed that mice who received MVAt produced higher levels of IFNγ, IL6 and IL2 in DLNs compared to MVAwt Conclusions: The deletion of interrelated immunomodulatory genes from MVA genome is a proper approach to improve its vaccine potential and it is a helpful tool that could contribute in the development of an HIV vaccine Vaccine-induced Intestinal and Salivary IgA Correlates with Reduced Viremia in Orallychallenged Neonatal Macaques Kara Jensen, Robert Wilson 2, Michael Piatak 3, Jeff Lifson 3, Uma Devi Ranganathan 4, William Jacobs, Jr. 4, Glenn Fennelly 4, Michelle Larsen 4, Koen Van Rompay 5, Pamela Kozlowski 2, Kristina Abel University of North Carolina at Chapel Hill and Center for AIDS Research, Chapel Hill, NC, United States, 2 Louisiana State University Health Sciences Center, New Orleans, LA, United States, 3 SAIC Frederick, Inc., Frederick, MD, United States, 4 Albert Einstein College of Medicine, Bronx, NY, United States, 5 CNPRC and UC Davis, Davis, CA, United States Background: HIV acquisition from breast milk is a significant route of pediatric HIV infections. Neonates produce less IgA than adults and are thus more susceptible to oral pathogens, including HIV. While serum antibodies are critical for controlling HIV post-exposure, the importance of mucosal IgA for the prevention of HIV transmission is not known. In addition, it is unclear how best to induce the development of specific mucosal antibodies through vaccination. Methods: We tested whether oral pediatric vaccination could induce SIV envelope (Env)-specific salivary and intestinal IgA, and protect against oral SIV acquisition. Neonatal rhesus macaques were mucosally immunized at birth with a live attenuated Mycobacterium tuberculosis strain engineered to express SIV Env and Gag, and twice boosted with MVAgpe. Animals were orally challenged beginning at week 9 using a weekly regimen of low-dose SIVmac25 (5000 TCID 50 ) to mimic HIV exposure from infected breast milk. Results: Vaccination induced SIV-specific plasma IgG and IgA antibodies in all animals by week 9, plus salivary and intestinal SIV-specific antibodies were detected in 8 of 8 and 3 of 8 animals, respectively. Although vaccination did not prevent infection, infants producing intestinal and salivary IgA (n=3, 37.5%) had significantly lower peak and set-point viremias compared to other vaccinated animals (p=0.009 and p=0.034, respectively). In addition, higher Env-specific mucosal IgA activities and plasma IgG avidities inversely correlated with peak viremia and viral set-point. Conclusions: Therefore, mucosal and systemic antibodies produced in response to neonatal oral vaccination contributed to controlled virus replication. Vaccine strategies that promote mucosal antibody development could help prevent oral HIV acquisition. 352 HIV Research for Prevention 204 HIV R4P

353 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.09 Construction and Evaluation of BCG and Modified Vaccinia Ankara Vaccines Expressing HIV- Subtype C Mosaic Gag Tsungai Ivai Jongwe, Ros Chapman, Shivan Chetty, Niki Douglass, Anna-Lise Williamson,2 University of Cape Town, Cape Town, South Africa, 2 National Laboratory Services, Cape Town, South Africa P4.0 ADCC Measurements in Rabbits Immunized with HIV- Vaccine Candidates Sanne S. Jensen, Marie Borggren, Gabriella Scarlatti 2, Leo Heyndrickx 3, Anders Fomsgaard, Ingrid Karlsson, NGIN consortium Statens Serum Institut, Copenhagen, Denmark, 2 San Raffaele Scientific Institute, Milan, Italy, 3 Institute of Tropical Medicine, Antwerp, Belgium Background: Over 90% of people with HIV- in sub-saharan Africa are infected with the clade C virus. High mutagenesis rates and diversity, even within clades, make it difficult to develop effective vaccines. Mosaic immunogens have been computationally designed to maximize inclusion of common T-cell epitopes. Compared to consensus immunogens, polyvalent mosaic immunogens of HIV- group M have increased breadth and depth of antigen-specific T-cell responses. Here, we determined the immunogenicity of vaccines expressing HIV- subtype C Gag mosaic in mice. Methods: BCG (BCG-Gag M ) and MVA (MVA-Gag M ) vaccines expressing the HIV subtype C mosaic gag gene were constructed. p24 ELISA and electron microscopy (EM) of MVA-Gag M -infected cells was used to determine the ability of the mosaic Gag to bud and form virus-like particles (VLPs). Shuttle vector integrity in BCG-Gag M was determined by PCR. Mice were primed intraperitoneally with 2x0 7 -cfu BCG-Gag M and boosted intramuscularly with 0 4 -pfu MVA-Gag M at week 0. Mice were sacrificed 2 days later and T-cell responses analysed by IFN-γ ELISPOT, CBA, and ICS assays. Results: Gag VLP production was confirmed by EM and p24 ELISA in the media of infected cells. A Th response was induced by the BCG- Gag M /MVA-Gag M vaccination regimen, and both CD4+ and CD8+ IFN-γ responses were detected. A potent effector memory phenotype was detected from both CD4+ and CD8+ cells. Overall, the BCG-Gag M prime MVA-Gag M boost induced robust HIV-specific CD4 + and CD8 + T cell responses that were 3 fold higher than responses induced by a control BCG prime MVA-Gag M boost. Genetic integrity of the BCG-Gag M vaccine was confirmed 0 weeks post vaccination in mice. Conclusions: These vaccines are immunogenic in Balb/c mice in a prime-boost vaccination regimen. Either vaccine alone had a dominant CD4 response while the combination of vaccines induced a greater CD8 response. The vaccines induce strong effector memory functions and will be further evaluated in non-human primates. Background: Induction of both neutralizing antibodies and nonneutralizing antibodies with effector functions e.g. antibody-dependent cellular cytotoxicity (ADCC) are desired in the search for effective vaccines against HIV-. In the search for novel immunogens capable of inducing an efficient antibody response, rabbits were immunized with selected antigens using different prime-boost strategies within the NGIN consortium. Methods: We selected 30 different groups of rabbits immunized with antigens from HIV- subtype A and B, including immunization with DNA alone, protein alone and DNA prime with protein boost. Rabbit sera were screened for ADCC activity using the GranToxiLux assay with human PBMC as effector cells and CEM.NKR CCR5 coated with rgp20 as target cells. This assay measures the proteolytic activity of Granzyme B after its delivery into target cells, initiated by antibody recognition of rgp20 on the target cell membrane. Results: The groups of rabbits with the highest serum ADCC activity were immunized with protein, monomeric gp20 or trimeric gp40, in CAF adjuvant. Interestingly, the ADCC activity did not correlate with neutralizing activity of purified IgG measured against SF62 and Bx08 with the pseudovirus-tzmbl assay, nor did it correlate with IgG gp20 ELISA binding titre. Conclusions: The antigens and/or immunization strategies capable of inducing antibodies with good ADCC activity do not necessarily induce good neutralizing activity and vice versa. When searching for an effective vaccine candidate it is important to evaluate the antibody response using an assay measuring the desired function

354 Posters Posters 4: Novel Vaccine and Prevention Concepts P4. Eliciting Cytotoxic T-lymphocyte Responses to HIV by Human Dendrocyte in vitro T-cell Activation with Synthetic Peptide-containing Microspheres Sarah S. Killingbeck,2, C V. Herst 3, Craig Rouskey 3, R M. Rubsamen 3 University of California, Division of Infectious Disease and Vaccinology, School of Public Health, Berkeley, CA, United States, 2 University of California, Immunology, Davis, CA, United States, 3 Immunity Project, Palo Alto, CA, United States P4.2 The Viral Vector Vaccine VSV-GP Boosts the Immune Response upon Repeated Applications Janine Kimpel, Reinhard Tober, Zoltan Banki, Lisa Egerer, Alexander Muik 2, Florian Kreppel 3, Dorothee von Laer Innsbruck Medical University, Division of Virology, Innsbruck, Austria, 2 Georg-Speyer-Haus, Frankfurt, Germany, 3 University of Ulm, Division of Gene Therapy, Germany Background: We describe a novel HIV vaccine consisting of synthetic d,l poly(l-lactic-co-glycolic) acid (PLGA) microspheres encapsulating HLA-specific cytotoxic T lymphocyte (CTL) HIV- Gag epitope KF in combination with CpG oligodeoxynucleotide (ODN) as adjuvant. We then demonstrate productive in vitro uptake and presentation of this peptide by healthy donor PBMC-derived dendrocytes (DCs), as well as the induction of specific CTL responses targeting KF when immunized DCs are co-cultured with autologous PBMCs. Methods: CTL responses to PLGA microsphere-encapsulated peptide seven days post incubation with autologous DCs presented with the microsphere-based vaccine in vitro were measured by interferon gamma (IFN-γ) ELISPOT. CTL killing of HIV-infected CD4+ T cells was quantified by p24 antigen ELISA specific for the p24 epitope within the HIV- Gag protein. Results: Antigen/peptide-experienced T cells are activated and secrete IFN-γ in response to peptide KF. In addition, CTLs elicited by microsphere-encapsulated peptides were capable of significantly restricting HIV replication in CD4+ T cells when compared to controls treated with microspheres containing OVA peptide. We observed a highly statistically significant (p< 0.000) reduction in CD4+ T cell viral load, approximately 40%, as assayed by p24 concentration in HIV-infected CD4+ T cells (from 026 pg/ml to 659 pg/ml) from one donor when co-cultured in 5: effector:target ratio with autologous antigen/peptide-experienced CD8+ T cells. Conclusions: Taken together, our experiments show a versatile new HIV vaccination method capable of eliciting robust HLA-specific CTL responses that efficiently control viral replication in vitro. This technique shows potential as a useful bioassay for a microsphere-based T cell vaccine with applications in manufacturing quality control and identification of a range of HLA types capable of presenting specific, HLA-restricted epitopes. Background: Vesicular stomatitis virus (VSV) is a potent candidate vaccine vector for various diseases. However, VSV s inherent neurotoxicity has limited its clinical application. Additionally, VSV induces neutralizing antibodies rapidly and is thus ineffective upon repeated applications. Our group has recently shown that VSV pseudotyped with the glycoprotein (GP) of the lymphocytic choriomeningitis virus, VSV-GP, is not neurotoxic. Here, we evaluated the potential of VSV-GP as a vaccine vector. Methods: We used Ovalbumin (OVA) as a model antigen and analyzed immunogenicity of GP-pseudotyped and wild-type VSV expressing OVA (VSV-GP-OVA and VSV-OVA) in vitro and in vivo in mouse models. Results: Mouse experiments revealed that both VSV-OVA and VSV-GP- OVA induced functional OVA-specific CTLs and anti-ova antibodies upon single immunization. However, boosting with the same vector was only possible for the GP-pseudotype but not for wild-type VSV. The efficacy of repeated immunization with VSV-OVA was most likely limited by the high levels of neutralizing antibodies, which we detected after the first immunization. In contrast, no neutralizing antibodies against VSV-GP were induced even after seven boost immunizations. CTL responses induced by VSV-GP-OVA were as potent as those induced by an adenoviral state-of-the-art vaccine vector. Additionally, immunization with both vectors completely protected mice from infection with Listeria monocytogenes expressing OVA. Conclusions: Taken together, VSV-GP is non-neurotoxic, induces potent immune responses, enables boosting and thus is a promising novel vaccine vector. 354 HIV Research for Prevention 204 HIV R4P

355 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.3 Single and Combined Vaccination Modalities Result in Distinct Immunological Profiles in HIV- gp40-immunised Mice P4.4 Intracellular Antigen Trafficking Directs Distinct Immune Responses Elicited by Dendritic Cell-targeting HIV Vaccines Deborah F.L. King, Paul F. McKay, Jamie F.S. Mann, Bryn Jones, Robin J. Shattock Imperial College London, Virology, London, United Kingdom Boon Kiat Lee, Jingying Zhou, Zhiwei Chen University of Hong Kong, AIDS Institute, Department of Microbiology, Hong Kong, Hong Kong Background: Recent studies have indicated that co-immunisation of protein antigens and DNA can induce both cellular and humoral immune responses. Methods: We investigated whether electroporated DNA and protein vaccinations by intranasal (IN) and intramuscular (IM) routes alone or as co-immunisations could be used to induce optimal HIV- gp40- specific responses in mice. Serum and vaginal antigen-specific IgG and IgA were measured by ELISA, and IFNγ+ T-cell responses were assessed by ELIspot. Results: IN Protein vaccination resulted in significantly higher antigenspecific IgG and IgA in the serum and IgA in the vagina, compared to DNA vaccination. IM protein vaccination resulted in similar levels of serum and vaginal IgG as IN vaccination. DNA vaccination induced significantly higher frequencies of gp40-specific IFNγ+ T-cells than IN or IM groups. Overall these results indicate that DNA vaccination effectively induces cellular IFNγ responses, but only low-level humoral responses, whereas IN vaccination optimally induces serum and vaginal antibody responses. DNA+IN co-immunisation resulted in significantly higher gp40- specific serum IgG than either DNA or IM vaccination, but similar levels to those observed with IN vaccination alone. The DNA+IN group also had significantly higher serum IgG and IgA than DNA+IM and DNA+IN+IM groups. The DNA+IN group exhibited significantly higher vaginal IgA compared to DNA, IM or DNA+IM groups, but again was not significantly higher than responses induced by IN vaccination alone. IFNγ-specific responses were similar in all co-immunisation groups, but significantly lower than DNA alone in DNA+IN, IN+IM and DNA+IN+IM groups. DNA+IN vaccination induced similar levels of IFNγ+ T cells as IN vaccination alone. Conclusions: Cellular and humoral HIV- gp40-specific responses were effectively induced by IN vaccination without the need for additional DNA co-immunisation. Thus this single vaccination route may be optimal for delivery of HIV vaccine antigens. Background: Improvement of HIV vaccine immunogenicity requires deeper understanding of antigen processing in dendritic cells (DC). To date, although antigen-targeting to DC has been explored as a novel strategy of HIV vaccine design, few studies have investigated the intracellular antigen processing and its influence on vaccine immunogenicity when different DC surface receptors are engaged. In this study, we conduct parallel experiments to determine the immunogenicity and possible underlying mechanism of DC-targeting HIV vaccines via the native ligands of PD- and CTLA-4, respectively. Methods: A panel of DNA fusion vaccines was constructed including soluble PD- (spd)-p24, soluble CTLA-4 (sctla-4)-p24, deletion mutant sδpd-p24, sδctla4-p24 and p24 alone. BALB/c mice were immunized with each of these DNA vaccines at 00µg dose via intramuscular in vivo electroporation following a prime/2-boost regimen with 3-week intervals. Post vaccination, immunogenicity profiles were analyzed using ELISA and ELIspot. Confocal microscopy was used to investigate p24 antigen trafficking to MHC-I or -II compartments in DC based on several endosomal markers. Results: DC-targeting HIV vaccines were significantly more potent than corresponding deletion mutant controls. spd-p24 is superior to CTLA4-p24 with following distinct immune responses: () significantly enhanced IgG2a (Th) antibody responses, (2) ~3.5-fold greater p24- specific CD8 + T cell responses besides a 2-fold increase in CD4 + T cells by IFN-γ ELISpots. Mechanistically, while p24 delivered by spd-p24 and sctla4-p24 routed to Lamp- endosomes for MHC-II presentation to CD4 + T cells, spd-p24 also routed to Rab4 endosomes for MHC-I cross-presentation to CD8 + T cell. Conclusions: PD- and CTLA-4 lead to distinct immunogenicity outcomes when used in our DC-targeting vaccines. Our findings provide evidence that the intracellular antigen processing in DC influences vaccine immunogenicity when different DC surface receptors are engaged

356 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.5 P4.6 Vaccines for Therapeutic Cellular Immunity that Target HIV Gag, Pol and Nef Epitopes to CD40 and DCIR Elicit T-cell Responses in Rhesus Macaques Yves Lévy,2,3, Sandra Zurawski,4, Anne-Laure Flamar,4, Christine Lacabaratz,2, Cécile Peltekian,2, Andres Salazar 5, Rodolphe Thiébaut,6, Gérard Zurawski,4 Vaccine Research Institute (VRI), Créteil, France, 2 Université Paris- Est Créteil Val de Marne (UPEC), Inserm U 955, Creteil, France, 3 Assistance Publique-Hôpitaux de Paris, Groupe Henri Mondor-Albert Chenevier, Service d Immunologie Clinique, Créteil, France, 4 Baylor Institute for Immunology Research, Inserm U 955, Dallas, TX, United States, 5 Oncovir, Washington DC, WA, United States, 6 INSERM U 897 Université Victor Segalen Bordeaux 2, Bordeaux, France Background: We have developed αcd40.hiv5pep and αdcir.hiv5pep, two dendritic cell (DC)-targeting vaccines, containing 5 T cell epitope-rich HIV regions of Gag, Pol, and Nef (G/P/N) directly fused to αcd40 (Flamar et. al., AIDS, 203) or αdcir mabs. In vitro, these vaccines expand multifunctional and polyepitopic specific CD4 + and CD8 + T cells. Methods: NHP were primed twice at weeks (W) 0 and 8 with MVA encoding G/P/N sequences encompassing the HIV5pep sequences and boosted with either αcd40.hiv5pep (n=6) or αdcir.hiv5pep (n=6) (three ID injections of 250µg with mg Poly-ICLC (Hiltonol) at W 2, 6 and 24). Two other groups of NHP received anti-dc vaccines at W 0, 4 and 2 (n=6 per group) followed by one MVA boost at W 22. Antibody and HIV5pep-specific T cell responses were detected by ELISA and IFNγ- ELISPOT performed two weeks after each immunization. Results: IgG titers were weak (< 200) or undetectable following the two MVA primes but were observed in all animals following the DC-targeting vaccinations (~5,000 /EC 50 ). Two weeks after each of DC-targeting vaccine boosts, HIV5pep-specific T cells were boosted to median 50, 503 and 388 SFC/0 6 PBMC for αdcir.hiv5pep and 350, 055 and 533 for αcd40.hiv5pep compared to after the second MVA vaccination [45 and 98 for these two groups]. Globally, DC vaccines boosted significantly T cell responses as compared to baseline (+648 SFC/0 6 PBMC, P<.0005). In non-mva primed groups, median HIV5pep-specific T cells responses following each DC-targeting vaccine were respectively 43, 35, 48 SFC/0 6 PBMC for αdcir.hiv5pep and 8, 90, 98 SFC/0 6 PBMC for αcd40.hiv5pep. Those responses increased to 200 and 30 SFC/0 6 PBMC two weeks after MVA boost. Conclusions: These DC-targeting vaccines elicited broad HIV peptidespecific T cell responses in both priming and boost settings. These data support their potential for further development, in particular as a component of a therapeutic vaccination strategy. Combined Approach of Vaccine and Antiviral Drugs Can Achieve Better Protection in NHP Model against Heterologous SHIV Challenges Ying Liu, Zhou Zhang, Yanling Hao, Shuhui Wang, Qiang Wei 2, Chuan Qin 2, Chang Liu, Hong Peng, Yiming Shao National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China, 2 Institute of Laboratory Animals Science, CAMS &PUMC, Beijing, China Background: Vaccine is needed to stop the AIDS pandemic. Even low efficacy vaccine can prevent millions HIV infections according to mathematic modelling. Unlike the rapid advances in antivirals since 990s and in biomedical interventions of recent years, HIV vaccine research has been struggling due to the unprecedented challenges it met. The best results of oral Prep of antivirals and RV44 vaccine showed around 50% and 30% protective efficacy respectively in clinical trials. The strategy of combining various prevention approaches, such as vaccines and antivirals, should be tested to achieve higher prevention. Methods: Rhesus macaques were immunized with DNA, recombinant vaccinia (replicating Tiantan strain) and protein vaccines expressing HIV- gag/pol/env and SIV gag. The immunized animals were given oral TDF/FTC before challenged by multiple low dose SHIV SF62 P3 through rectal mucosal. Immune responses and viral load were monitored for evaluation. Results: Among the 4 groups of monkeys (8 per group), high peak viral loads (3.8 x 0 6 ) can be detected in all animal of the control group and reduced peak viral load were detected in 4 (.7x 0 5 ), 3 (2.9x 0 3 ) and (x 0 3 ) animals of the vaccine only, antiviral only and vaccine and antiviral combined groups respectively. The reduced viral load in the three treated groups also resulted in low viral set-point and faster felling to undetectable levels, compared to the control group. Both strong humoral (binding and VV2 antibodies) and cellular (CD8 and CD4) responses to HIV- antigens and SIV gag can be detected in immunized animals. No viral load can be detected in the protected animal after inoculation them with anti-cd8 antibody. Conclusions: High protective efficacy (87.5%) can be achieved by the combined use of vaccines and antivirals. Experiments are pending to evaluate the mechanism of protection in protected animals, as well to the impact of the treatment on the viral reservoirs in the infected animals. 356 HIV Research for Prevention 204 HIV R4P

357 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.7 Longitudinal Analysis of SIVmac239 Mutations around the 2 Protease Cleavage Sites and their Correlations with Viral Load Reduction and CD4 counts Ma Luo,2, David Tang, David La, Rupert Capina, Xin-Yang Yuan 3, Jorge Correia-Pinto 4, Cecilia Prego 4, Maria Alonso 4, Christina Barry 3, Richard Pilon, Christina Daniuk, Mikaela Nykoluk, Stephane Pillet 3, Tomasz Bielawny, Jeffrey Tuff, Chris Czarnecki, Philip Lacap, Gary Wong 3, Shaun Tyler 3, Ben Liang 3, Zhe Yuan 5, Qingsheng Li 5, Terry Blake Ball, Paul Sandstrom, Gary Kobinger 3, Francis Plummer 2,3 National Microbiology Laboratory, National HIV and Retrovirology Laboratory, Winnipeg, MB, Canada, 2 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada, 3 National Microbiology Laboratory, Winnipeg, MB, Canada, 4 University of Santiago de Compostela, Santiago de Compostela, Spain, 5 University of Nebraska- Lincoln, Nebraska-Lincoln, NE, United States P4.8 Sequences Surrounding the 2 Protease Cleavage Sites are Good Targets for Both Prophylactic and Therapeutic HIV Vaccines David Tang, David La, Rupert Capina, Xin-Yang Yuan, Jorge Correia-Pinto 2, Cecilia Prego 2, Maria Alonso 2, Christina Barry 3, Richard Pilon, Christina Daniuk, Mikaela Nykoluk, Stephane Pillet 3, Tomasz Bielawny, Jeffrey Tuff, Chris Czarnecki, Philip Lacap, Gary Wong 3, Shaun Tyler 3, Ben Liang 3, Zhe Yuan 4, Qingsheng Li 4, Terry Blake Ball, Paul Sandstrom, Gary Kobinger 3, Francis Plummer 3,5, Ma Luo,5 National Microbiology Laboratory, National HIV and Retrovirology Laboratory, Winnipeg, MB, Canada, 2 University of Santiago de Compostela, Santiago de Compostela, Spain, 3 National Microbiology Laboratory, Winnipeg, MB, Canada, 4 University of Nebraska-Lincoln, Nebraska-Lincoln, NE, United States, 5 University of Manitoba, Medical Microbiology, Winnipeg, MB, Canada Background: HIV- protease mediates the cleavage of Gag, Gag-Pol and Nef precursor polyproteins in a highly specific and temporally regulated manner. Because a total of 2 cleavage reactions are required to generate a mature virion, generating focused immune response targeting the sequences surrounding the protease cleavage sites (PCS) could drive viral mutations to its disadvantage. We have conducted a proof of concept study with Cynomolgus macaques and pathogenic SIVmac239 as a model and used a modified recombinant vesicular stomatitis vector and nanocarriers to deliver 2 20-amino acid antigens. We showed that a vaccine targeting the sequences surrounding the 2 protease cleavage sites is promising at prevention of HIV- infection and disease progression. In this study we systematically analyzed breakthrough viruses of vaccinated and control macaques. Methods: The sequences surrounding the 2 protease cleavage sites were amplified from plasma RNA of all SIVmac239 positive samples. The amplified PCR products were sequenced with 454 pyrosequencing technology. The amino acid and frame shift mutations were analyzed and correlated with viral load and CD4 counts. Regression analysis was conducted to correlate the viral mutations with viral load and CD4 counts. WebLogo was used to plot the amino acid mutations. Results: Extensive mutations were detected around PCS and both conserved and non-conserved mutations are correlated with lower viral load (p< 0.000). The breakthrough viruses from the vaccinated macaques carry significantly higher mutations than the controls. Longitudinal analysis revealed that the high rate of non-conserved and conserved amino acid mutations along the sequences surrounding the PCS lead to the reduction and diminishing of viral load. Conclusions: The pathogenic SIVmac239 is extremely vulnerable to any amino acid alternations around PCS and focusing immune response to sequences surrounding the PCS of HIV- can drive amino acid mutations and lead to complete viral control. Background: HIV- protease mediates the cleavage of Gag, Gag-Pol and Nef precursor polyproteins in a highly specific and temporally regulated manner. Because a total of 2 cleavage reactions are required to generate a mature virion, generating focused immune response targeting the sequences surrounding the protease cleavage sites (PCS) could drive viral mutations to its disadvantage. We have conducted a pilot study to investigate the feasibility and effectiveness of a vaccine targeting the sequences around the 2 PCS using Cynomolgus macaques and pathogenic SIVmac239 as a model. Methods: Twelve recombinant VSVpcs were used to immunize 2 Cynomolgus macaques and nanopackaged PCS peptides were used as a boost. The immunized macaques and 6 controls were repeatedly challenged intrarectally with an increased dosage of SIVmac239. Antibody and T cell responses to the PCS peptides, CD4+ and CD8+ T cell counts and challenge dosage were monitored. 454 Pyrosequencing was conducted to analyze break-through viruses and the amino acid mutations surrounding the PCS sites were correlated with viral load. Results: Antibody and T cell responses to the 2 PCS protected macaques against higher dosage of SIVmac239 intrarectal challenge (p=0.005, R=0.42). The vaccine group maintained higher CD4+ counts (p=0.0002) than the controls weeks after being infected. Analysis of viral mutations around 2 PCS of 276 samples (4 to 20 sampling points/monkey) detected extensive mutations. These mutations, both conserved and non-conserved amino substitutions around PCS, are correlated with lower viral load (p< 0.000). Conclusions: Our study with nonhuman primates and pathogenic SIVmac239 as a model showed that a vaccine targeting the sequences surrounding the 2 protease cleavage sites is promising at prevention of HIV- infection and disease progression. It demonstrated that the pathogenic SIVmac239 is extremely vulnerable to any amino acid alternations around PCS. Targeting PCS of HIV- could be an effective vaccine approach

358 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.9 P4.20 Antibodies to CD52g, a Secreted Spermcoating Antigen, Agglutinate Seminal Leukocytes and Prevent their Infiltration into Vaginal Epithelium Jai G. Marathe, Joseph Politch 2, Ayesha Islam 2, Kevin Whaley 3, Thomas Moench 4, Deborah J. Anderson 5 Boston University, Medicine, Boston, MA, United States, 2 Boston University, Obstetrics and Gynecology, Boston, MA, United States, 3 Mapp Biopharmaceutical, Inc., San Diego, CA, United States, 4 ReProtect Inc., Baltimore, MD, United States, 5 Boston University, Obstetrics and Gynecology, Medicine, Microbiology, Boston, MA, United States Background: Our program is studying the topical use of monoclonal antibodies (mabs) for contraception and HIV prevention. mabs directed against CD52g, an antigen secreted into the male genital tract and inserted into sperm membranes, potently agglutinate sperm and are being developed for contraceptive use. We are also seeking strategies to prevent cell-associated HIV transmission mediated by HIV-infected seminal white blood cells (swbc). In this study, we investigated whether CD52g also attaches to swbc, and whether anti-cd52g mabs agglutinate these cells and/or inhibit their interaction with the vaginal epithelium. Methods: Dylight 633-conjugated MSH8, a mouse anti-cd52g mab (gift of J. Herr), was used in flow cytometry experiments to detect CD52g on swbc and passive insertion of seminal plasma CD52g into the plasma membrane of monocyte-derived macrophages (MDMs). HC4, a human mab expressed in Nicotiana (Mapp Biopharmaceuticals Inc.), was used in two functional assays: ) agglutination of swbc, assessed by counting the percentage of CMFDA-labelled WBC associated with sperm agglutinates in mab-treated seminal fluid; and 2) cell attachment and infiltration assays, modeled with CD52g-coated CMFDA-labeled WBCs and EpiVaginal tissue (MatTek Corp.), and assessed by confocal microscopy. Results: MSH8 bound to a majority of swbcs and also to seminal plasma-treated MDMs. HC4 trapped WBCs in sperm agglutinates. The antibody also significantly inhibited the attachment and infiltration of CD52g-coated WBCs into the vaginal epithelium. Control mabs had no effect in these assays. Conclusions: Topical vaginal application of antibodies to CD52g could serve a dual purpose use: prevention of pregnancy and inhibition of cellassociated HIV transmission. Vaccine-induced CD07a + CD4 + T-cells Are Resistant to Killing Following Immunodeficiency Virus Infection Tetsuro Matano,2, Kazutaka Terahara, Hiroshi Ishii, Takushi Nomura National Institute of Infectious Diseases, AIDS Research Center, Tokyo, Japan, 2 University of Tokyo, Institute of Medical Science, Tokyo, Japan Background: CD4 + T cell responses are crucial for effective antibody and CD8 + T cell induction following virus infection. However, virus-specific CD4 + T cells can be preferential targets for HIV infection. HIV-specific CD4 + T cell induction by vaccination may thus result in enhancement of virus replication following infection. We examined virus-specific CD4 + T cell responses before and after SIV infection in vaccinated macaques and the sensitivity of vaccine-induced CD4 + T cells to killing following infection. Methods: We analyzed SIV-specific T-cell responses using frozen PBMCs from vaccinated (n = 8) and unvaccinated (n = 2) Burmese rhesus macaques. Vaccinated animals received a DNA prime and a Gag-expressing Sendai virus vector boost, followed by an intravenous SIVmac239 challenge (3 months post-boost). Eleven (6%) of vaccinated animals controlled SIV replication. SIV-specific CD4 + T-cell responses were examined at or 2 months pre-challenge and week post-challenge by measurement of five markers, CD07a, MIP-β, IFN-γ, TNF-α, and IL-2 following SIV-specific stimulation. Results: At week post-challenge, both vaccinated and unvaccinated animals had higher frequencies of SIV-specific CD4 + T-cell subsets inducing CD07a than other markers. Comparison of virus-specific CD4 + T-cell responses in vaccinated animals pre- and -week post-challenge showed a significant reduction in CD07a-negative but not in CD07a + subsets after SIV challenge. The reduction in vaccinated non-controllers was larger than controllers. Conclusions: Vaccine-elicited CD4 + T cells with the potential to induce CD07a are resistant to infection in a macaque AIDS model. Our results indicate that vaccine-induced CD07a-negative CD4 + T cells are efficiently killed following exposure, suggesting that vaccine design for HIV protection should avoid CD07a-negative CD4 + T-cell induction. 358 HIV Research for Prevention 204 HIV R4P

359 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.2 Factors that Influence the Willingness of Young Adults to Participate in Early Vaccine Trials and Contraceptive Practices in Dar es Salaam, Tanzania P4.22 Evaluation of Dendritic Cell Targeted Consensus B and MOSAIC HIV Gag Protein Vaccines in Vitro in PBMC of Treatment Naive HIV- Infected People Theodora Mbunda, Muhammad Bakari, Edith Tarimo, Eric Sandstrom 2, Asli Kulane 2 Muhimbili University of Health and Allied Sciences, Dar es Salaam, Tanzania, United Republic of, 2 Karolinska Institutet, Stockholm, Sweden Background: HIV/AIDS destroys the lives of young people especially in low-income countries. The inclusion of youths in HIV vaccine trials is necessary in obtaining an effective vaccine that is acceptable to them. Participation in HIV vaccine trials however necessitates participants not to become pregnant or impregnate women. Therefore it is important to study factors influencing willingness to participate and dynamics of contraceptive practices among young adults Methods: Four hundred and fifty young adults visited a youthfriendly Infectious Diseases Clinic (IDC) from February to September 202 completed a self-administered questionnaire concerning sociodemographic information, contraceptive practices, knowledge and perception of HIV vaccine studies, and the availability of social support. Results: Of our participants, 50.6% expressed willingness to participate. The willingness was positively correlated with having some knowledge about HIV vaccine studies (AOR, 2.2; 95% CI:.4-3.4), a positive perception toward such studies (AOR, 2.3; 95% CI:.5-3.6) having a relationship with someone who could help them make a decision (AOR, 2.5; 95% CI:.3-4.9), and age at the time of sexual debut (AOR, 2.6; 95% CI.0-6.7) for 5- to 9-year-olds and (AOR, 2.7; 95% CI.0-7.) for older participants. 73% of those expressing WTP knew contraception was for spacing children, 45% and 49% reported to ever use contraceptives and to use them at time of last sexual intercourse respectively. The reasons for not using contraceptives were not being married; lack of knowledge on contraceptives; and having unplanned sexual intercourse. Conclusions: The participants exhibited a moderate willingness to participate in HIV vaccine trials, less than half reported to use contraceptives. More efforts should be made to inform the youths about specific HIV vaccine trials, to engage significant others in the decisionmaking process, and to address impediments pertaining to contraceptive use in the Tanzanian context. Godwin Nchinda, Nadesh Nji 2, Jules C Tchadji 2, Georgia Ambada 2, Andres Salazar 3, Ralph Steinman 4, Michel Nussensweig 4, Tibor Keller 5, Samuel Sosso 2, Chae Gyu Park 6, Pierre Fouda 7, Vitorro Colizzi 8 Chantal Biya International Reference Center for HIV, Microbiology and Immunology, Yaounde, Cameroon, 2 Chantal Biya International Reference Center for HIV, Yaounde, Cameroon, 3 Oncovir, Jonesboro, GA, GA, United States, 4 Rockefeller University, New York, NY, United States, 5 Celldex Therapeutics, Hampton, NJ, United States, 6 Yonsei University College of Medicine, Seoul, Korea, Democratic People`s Republic of, 7 Central Hospital of Yaounde, Yaounde, Cameroon, 8 Tor Vergata, Roma, Italy Background: Our consortium has developed a first generation dendritic cell (DC) targeted consensus B HIV gag protein vaccine for proof of concept studies that selected delivery of proteins to DC will allow proteins to be more immunogenic. This should provide a cheaper and effective way to immunize people repeatedly with no negative impact of pre-existing immunity. This vaccine is already in an ongoing phase clinical trial in New York however it is not known whether this consensus B based vaccine would work in sub Saharan African where unrelated strians of HIV- are predominant. Methods: To assess the consequense of targeting a consensus B and MOSAIC gag protein vaccines to DC from people infected with unrelated strains of HIV- in Africa we added the protein vaccines to their blood cells in vitro and measured proliferation and IFNy production by bulk PBMCs as well as a coculture between monocyte derived DC and T cells. Results: Dendritic cell targeted Consensus B and MOSAIC HIV gag Protein vaccines recalled pre-existing T cell responses in blood cells of treatment naïve people infected with unrelated strains of HIV-. DC targeted MOSAIC gag vaccine was more efficient than the consensus B vaccine and stimulated significant proliferation of HIV gag specific cells (P< 0.00). Conclusions: Thus dendritic cell targeted consensus B and MOSAIC HIV gag protein vaccines could recall pre-exhisting CD4 and CD8 T cell responses in vitro in PBMC of treatment naïve people infected with unrelated strains of HIV- in AFrica. DC targeted MOSAIC gag protein vaccine should be further evaluated for possible incorporation in future vaccines

360 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.23 P4.24 Gp4 and p24 IgG Binding Antibody Titers Are Associated with HIV- Viral Control during Early HIV- Subtype C Infection Bongiwe G. Ndlovu, Amy Chung 2, Jennifer Maroa 3, Bruce D. Walker,2, Galit Alter 2, Thumbi Ndung u,3 HIV Pathogenesis Programme, University of KwaZulu-Natal, Durban, South Africa, 2 Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States, 3 KwaZulu-Natal Research Institute for Tuberculosis and HIV, University of KwaZulu-Natal, Durban, South Africa Background: The RV44 Thai HIV- vaccine trial showed 3% protective efficacy and suggested a role for non-neutralizing antibodies in protection against risk. However, the kinetics of development of binding antibodies and their ability to mediate innate effector functions in HIV- infection has not been fully characterized. The purpose of this study was to characterize the evolution of anti-hiv binding antibodies and to determine their antibody-dependant cellular phagocytic activity (ADCP) in individuals with recent HIV- infection. Methods: Thirty four individuals with acute HIV- infection (Fiebig I/ II) were identified and followed for one year in Durban, KwaZulu-Natal. Viral loads were measured using the Roche Amplicor ver.5 assay. Plasma HIV- specific gp20, gp4 and p24 total IgG and Ig subclasses (IgG-4) antibodies were measured using a customized Multiplex Luminex assay at, 2, 3, 6 and 2 months post-infection. The capacity of plasma antibodies to mediate ADCP in the presence of fluorescent beads conjugated to gp20 protein was examined in a THP- cell based in-vitro assay. Results: Total gp20 and p24-specific IgG antibody titers significantly increased over the follow up period (p< 0.000, One-way ANOVA), whereas gp4-specific IgG titers did not change over time compared to titers at one month post-infection. Both gp4 and p24-specific total IgG antibodies correlated negatively with contemporaneous viral load at months 3 (p= 0.06, r=-0.38; p=0.046, r=-0.457) and 2 (p= , r=-0.465; p=0.0524, r=-0.439) post-infection. Despite high IgG binding titers, neither anti-gp20, gp4 nor p24 IgG titers correlated with viral load. Gp20-specific phagocytic scores correlated with higher viral set point. Conclusions: The kinetics of expansion of HIV- protein-specific IgG titers were different. Gp4 and p24-specific IgG titers may have an anti- HIV role in vivo which requires further investigation. Host Restricted Poxviruses Produce Distinct Host Responses in an in vivo Mouse Model with Implications for Future Use as HIV- Vaccine Vectors Kristy Offerman, Olivia Carulei, Armin Deffur 2, Nicola Douglass, Anna-Lise Williamson,3,4 University of Cape Town, Division Medical Virology, Department of Clinical Laboratory Sciences, Cape Town, South Africa, 2 University of Cape Town, Clinical Infectious Diseases Research Initiative, Cape Town, South Africa, 3 Institute of Infectious Disease and Molecular Medicine, Cape Town, South Africa, 4 National Health Laboratory Service, Groote Schuur Hospital, Cape Town, South Africa Background: Host restricted poxviruses make promising candidates for HIV vaccine vectors due to their safety profile and immunogenicity. The comparison of host responses produced by different poxvirus vectors would aid in the assessment and rational design of improved vaccines. Methods: In order to investigate the ability of different poxviruses to interact with the mammalian host, we compared host gene expression profiles in the spleens of BALB/c mice in response to infection (05 pfu/ mouse) with four poxvirus species; Lumpy Skin Disease virus (LSDV), Canarypox virus (CNPV), Fowlpox virus (DCEP 25 modified strain (Merial)) (FWPV), modified Vaccinia Ankara (MVA). Results: The findings presented here indicate that four, genetically diverse host-restricted poxviruses, CNPV, FWPV, MVA and LSDV, produce qualitatively and quantitatively distinct host responses in an in vivo mouse model. Differential gene expression regulated by the different poxviruses is discussed with particular emphasis on immunityrelated responses. Of particular interest, CNPV and FWPV, and not MVA or LSDV induce the upregulation of two immunoglobulin genes (Ighg and Ighg3 (IgG3)) with CNPV inducing a third, Ighm (IgM). Comparison of the immune responses resulting from the partially effective clinical RV44 HIV- trial and the ineffective VAX003 trial indicated that HIV--specific IgG3 antibodies correlated with decreased risk of HIV- infection in the RV44 trial. The upregulation of IgG3 by avipoxviruses in vivo, suggest that the avipoxvirus-vector backbone may be involved in stimulation of the clinically important IgG3 antibody subclass. Conclusions: These findings suggest potentially important biological differences in response to four clinically relevant poxviruses, especially because small changes in gene expression may produce disproportionate results in biological systems. These differences may affect the immune response induced to vaccine antigen in vectors based on these viruses. 360 HIV Research for Prevention 204 HIV R4P

361 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.25 P4.26 Generating an Anti-HIV Vaccine Using Nucleoside-modified mrna Encoding Envelope Norbert Pardi, Katalin Kariko, Michael Hogan, Hiromi Muramatsu, James A. Hoxie, Drew Weissman University of Pennsylvania, Department of Medicine, Philadelphia, PA, United States Background: There has been great progress in understanding the detailed molecular mechanisms of HIV- infection but no effective vaccine has been developed to date. mrna has emerged as a very promising new therapeutic agent in recent years and has already shown progress as an effective method for generating potent vaccines. Methods: To create a vaccine with maximal potency, in vitro transcribed mrnas were optimized for higher levels and extended translation by incorporation of selected UTRs, modified nucleosides, 5 cap, 3 poly(a)- tail, and other modifications and were HPLC-purified. To achieve both strong T cell and B cell responses, heterologous mrna prime - protein boost vaccination regimens were used. For priming, naked mrna encoding HIV envelope gp60 was administered intradermally into mice. Cell surface Env was used to increase exposure of neutralizing epitopes. Four weeks after the second mrna prime, Env protein was injected intramuscularly as a boost. As nucleoside modified mrna does not activate RNA sensors, to increase the robustness of the immune response, a series of adjuvant molecules and encoding mrnas were co-injected along with the antigen-encoding mrna. Flow cytometry and ELISA were used to evaluate T cell and B cell responses, respectively. Results: Elevated levels of IFN-χ, TNF-α and IL-2 in antigen-specific CD4 + and CD8 + T cells and high gp20 antibody titers could be measured following two rounds of mrna prime - protein boost vaccination. Conclusions: Our results demonstrate that antigen-encoding nucleoside modified mrna induces effective HIV-specific immune responses and has great potential for vaccination against infectious diseases. Comparative Neutralization Sensitivity of Indian and South African HIV- Clade C Viruses to Plasma Antibodies from Chronically Infected Indian Donors Shilpa Patil, Sharda Gadhe 2, Swapnil Sonawane 2, Manish Bansal, Suprit Deshpande, Tandile Hermanus 3, Lynn Morris 3, K G Murugavel 4, Suniti Solomon 4, Seema Sahay 2, Ramesh Paranjape 2, Bimal K. Chakrabarti, Jayanta Bhattacharya HIV Vaccine Translational Research Laboratory, THSTI-IAVI HIV Vaccine Design Program, Gurgaon, India, 2 National AIDS Research Institute, Pune, India, 3 National Institute for Communicable Diseases, Johannesburg, South Africa, 4 Y.R. Gaitonde Centre for AIDS Research and Education, Chennai, India Background: HIV- clade C is the major subtype circulating in India and South Africa. The present study was undertaken to examine the extent of neutralization sensitivity of Indian and South African (SA) HIV- clade C viruses to plasma antibodies obtained from anti-retroviral naïve chronically infected HIV- positive Indian patients. Methods: Plasma samples from 50 anti retroviral naïve donors from India chronically infected with HIV- were assessed for their degree of neutralization of 9 Indian and 0 South African HIV- clade C envelopes using a TZM-bl reporter cell assay. Reagents were shared between Indian and SA laboratories for quality assessment. Antibody specificities were determined by using TriMut core protein, mutant and chimeric viruses. Results: 27/50 (8%) plasma samples were found to neutralize >50% of the panel viruses at :00 dilution. Amongst these, seven were found (4.66%) displayed maximum breadth and potency with median ID50s ranging from The BCN plasma antibodies were found to neutralize Indian (57%) viruses slightly better than SA (43%) viruses. None of the potent broadly cross neutralizing BCN plasmas was found to show neutralizing antibodyspecificities targeting thecd4 binding site. However, two of them showed N332 residuedependence in V3 loop in both Indian and SA clade C Env backbones. While 3/7 BCN plasma antibodies showed clade C MPER (HIV-2/HIV 732-CC) dependence, they did not show specificity to epitopes targeted by known MPER directed monoclonal antibodies. Conclusions: Our data suggest the presence of common epitopes in Indian and South African HIV- clade C envelopesas most of the BCN plasma antibodies cross-neutralized pseudotyped viruses expressing clade C envelopes from both the countries. Identification of epitopes common in both in Indian and SA clade C envelopes will help define strategies to design vaccine that would be effective in both countries. 36

362 Posters Posters 4: Novel Vaccine and Prevention Concepts P4.27 Cell-surface Display and Panning of HIV- Derived Envelope Proteins P4.28 Development of Advanced Oligonucleotidebased Microbicides: Driving HIV into Suicide Tim-Henrik Bruun, Veronika Schmid, Alexander Kliche, Ralf Wagner University of Regensburg, Institute of Medical Microbiology and Hygiene, Molecular Microbiology and Gene Therapy Unit, Regensburg, Germany Maike Voges, Joachim Hauber, Karin Moelling,2 Heinrich-Pette Institute, Antiviral Strategies, Hamburg, Germany, 2 Max Planck Institute for Molecular Genetics, Berlin, Germany Background: Available display systems allow the screening of millions of candidate proteins and are the method of choice to identify optimized antigen-antibody binding. We established a mammalian cell-surface display to present HIV- envelope derivatives in a natural, trimeric and membrane bound environment. This allows us to generate affinity enhanced envelope derivatives against broadly neutralizing antibodies (bnabs) in order to select potent Envelope (Env) based vaccine candidates. Methods: An HIV- derived lentiviral vector was developed to infect HEK293T cells at a low multiplicity of infection (MOI), in order to correlate phenotype and genotype. The vector was designed and proven to express both GFP and Env in a constant relationship, enabling indirect normalization for Env expression by detecting GFP. After staining with an appropriate bnab, Env-displaying cells were selected for high affinity binding via FACS-Sorting. To adapt this system to the use of very large libraries, a refined vector system was developed, allowing the stable genomic integration of both ENV and GFP at a distinct integration site. This ensures that only one envelope variant is expressed per cell, efficiently linking phenotype and genotype. Due to the stringent linkage of ENV and GFP, GFP expression can again be used as a means to normalize for Env expression in the FACS-Sorting process. Results: A small model library of Env variants with distinct binding capacities towards the bnab D was used to evaluate both techniques. The proof-of-concept experiment for the virus-based approach led to an enrichment of up to tenfold for the Env variant with the highest affinity toward D after two rounds of selection. Using the stable cell line technology, the desired Env high affinity variant could even be enriched up to 39-fold after only one round of FACS-Sorting. Conclusions: Hence, these techniques provide the possibility to screen for membrane bound Env variants with high binding capacities towards any bnab applied. Background: HIV is transmitted primarily by sexual intercourse and predominantly infects people in third world countries. An important medical need is self-protection of women, particularly in societies where condoms are not accepted. Therefore, we currently develop an oligodeoxynucleotide (ODN)-based microbicide, which destroys HIV prior to infection by activating the retroviral RNase H. Methods: We performed RT/RNase H cleavage assays in vitro to demonstrate RNase H-dependent cleavage of viral RNA in the highly conserved polypurin tract after incubation with ODNs. Furthermore, luciferase-based infection assays and p24 ELISA were used to analyze de novo infection capacity by ODN-treated HIV. Finally, various ODN stability assays were performed. Results: It is shown that ODNs target cell-associated as well as cell-free virions, leading to efficient degradation of the viral RNA genome and, in turn, to strongly reduced infectivity. Moreover, pronounced antiviral activity is demonstrated in vitro and in vivo (animal) models. Surprisingly, ODNs are able to enter cell-free virions without any carrier and are also stable over weeks at 37 C in different solutions (PBS, BSA, KY Jelly). Conclusions: ODNs trigger highly effective destruction of the viral RNA genome prior to infection without any carrier and are highly stable for many weeks. Therefore, ODNs may be valuable components of advanced microbicides to drive HIV into suicide. 362 HIV Research for Prevention 204 HIV R4P

363 Posters 4: Novel Vaccine and Prevention Concepts Thursday, 30 October P4.29 P4.30 LB Induction of Broadly Neutralizing Antibodies to HIV- via DNA Motif Immunization Xilin Wu, Zhiwei Wu 2, Lin Cheng 3, Zhiwei Chen 3 The University of Hong Kong, Hong Kong, Hong Kong, 2 Nanjing University, NanJing, China, 3 The University of Hong Kong, HongKong, Hong Kong Background: HIV- is highly mutated, which poses a serious problem to develop a prophylactic AIDS vaccine. To date, it remains elusive how to induce broadly reactive neutralizing antibodies against genetically divergent HIV- strains. We have previously invented peptide motif immunization, which could induce broadly neutralizing antibodies against highly conserved and functional domain of HIV- envelope protein in mice, rabbits and rhesus monkeys. In this study, we aim to invent a novel DNA motif immunization method. Methods: The primers encoding 5 amino acids GPG motif were synthesized, which were inserted in the plasmid by PCR as the gene of motif. We constructed four groups of plasmid in the vector of Pvax, PD-p24- motif, PD-ova-motif, CTLA4-p24 and PD-p24 as control. Mice received 4 DNA immunizations by intramuscular injection plus electroporation. Two weeks after the final immunization, ELISA, Neutralization, Elispot and FACS were conducted to test the efficacy of such antigen in the responses of humoral immunity and cellular immunity. Results: The plasmid encoding the motif gene was characterized by sequencing. It showed that the motif gene was constituted by nine fixed nucleotides encoding three amino acid residues of GPG in the center flanking with 36 random nucleotides. Plasmid expression was confirmed by FACS. The plasmid containing the gene of motif all could induce antibody specific for GPG with comparing to the control of PD-p24 group. In addition, the group of PD-p24-motif could induce GPG-specific antibodies in mice that neutralized 00% HIV- strains of various subtypes tested with GPG motif in their V3 region. Moreover, it could provide robust cellular immunity against gag protein of HIV. Conclusions: Our findings firstly completed proof of concept of DNA motif immunization. What s more, the PD-p24-motif plasmid could be a potential candidate DNA vaccine against HIV infection with induction of broadly neutralizing antibodies and robust cellular immunity. IL-R, WNT and γ-c Cytokine Receptor Signaling Correlate with CD4 T Cell Effector Function in Response to Anti-DEC205-HIV gag p24/polyiclc Vaccination Khader Ghneim, Marina Caskey 2, Christine Trumpfheller 2, Gaelle Breton 2, Petra Stafova, Stephanie Richards, Richard Koup 3, Bob Bailor 4, Sarah Schlesinger 2, Jeff Ahlers, Rafick P. Sekaly 5, Ralph Steinman 2, Mark Cameron Vaccine & Gene Therapy Institute of Florida, Port Saint Lucie, FL, United States, 2 Rockefeller University, New York, NY, United States, 3 Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States, 4 Vaccine Research Center, National Institute of Allergy and Infectious Diseases Vaccine Immune T-Cell and Antibody Laboratory, National Institutes of Health, Bethesda, MD, United States, 5 Case Western Reserve University, Cleveland, OH, United States Background: A randomized double-blinded, placebo-controlled phase I study was conducted in healthy volunteers to evaluate the safety and immunogenicity of the dendritic cell targeted mab (3G9) fusion protein vaccine, anti-dec205-hiv gag p24 with polyiclc (DCVax). Subjects were enrolled in three dose arms: Low(0.3mg), Mid(mg) or High(3mg) with.6mg of polyiclc subcutaneously. Control arms received either polyiclc or Saline. Methods: We performed microarray and bioinformatic analysis to identify gene signatures and pathway activity differentially expressed in whole blood from the different vaccine dose groups. Linear Regression analysis was used to correlate gene expression with ICS data. Statistical Inference was assessed using Wilcoxon rank sum test. Results: DCVax induced greater IFN gene expression and IL-2 production in whole blood compared to polyiclc alone, and in a dosedependent fashion that correlated with PRDM(BLIMP) expression in CD4 cells and total Gag-specific CD4 TH and CD8 IFNγ producing T cells in the high dose vaccinees. Pathway analysis on top regression features showed differential proinflammatory responses (e.g. IL, RXR, LPS) at d in subjects that mounted potent Gag-specific CD4 responses. High dose subjects upregulated genes downstream of IL-R signaling (e.g. IRAK3, MAPK2K3) including NF-κB. Gag-specific CD4 TH responses correlated with ILR, WNT, and γ-c receptor cytokine signaling pathways that might also predispose toward greater CD4 T cell memory development. Conclusions: Synergy was seen with the high dose HIV Gag p24 dendritic cell targeted vaccine and polyiclc for induction of both a quantitative and qualitative greater Type I IFN response compared to lower vaccine doses or polyiclc alone. This resulted in optimum CD4 TH and CD8 CTL responses. Of note, only the high dose and polyiclc combination induced SOCS-, a critical regulator of type I interferon signaling. These results highlight that combinations of vaccine and adjuvant can trigger quantitatively different responses

364 Posters Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42.0 P42.02 Self-reported Benefits of Study Participation in HVTN 503, Phambili Mary A. Allen, Barbara Metch 2, Zoe Moodie 2, Linda-Gail Bekker 3, Gavin Churchyard 4, Koleka Mlisana 5, Maphoshane Nchabeleng 6, Jim Kublin 2, Glenda Gray 7, HVTN 503 Study Team NIH/NIAID, DAIDS, Bethesda, MD, United States, 2 FHCRC, Seattle, WA, United States, 3 Desmond Tutu HIV Foundation, Cape Town, South Africa, 4 Aurum Institute, Klerksdorp, South Africa, 5 University of KwaZulu Natal, Durban, South Africa, 6 Medunsa Clinical Research Unit, Medunsa, South Africa, 7 Chris Hani Baragwanath Hospital, PHRU, Soweto, South Africa Background: Participants (ppts) in HVTN 503 ( Phambili ), the only HIV vaccine efficacy trial conducted in Africa to date, were asked about possible benefits of study participation. Methods: Social impact (SI) assessment was conducted at weeks 2, 78, 30, and 82, and included asking In the last 6 months, has participation in this study had a beneficial impact on your life? Benefits were analyzed by sex, age, study site and treatment group. Results: Of the 80 ppts enrolled, 752 (94%) reported that study participation had a beneficial impact on their lives, and only 48 (6%) reported negative social impacts (NSI). Overall, 705 (88.0%) reported a benefit and no NSIs; 48 (6.0%) reported neither; and reported a NSI and no benefit. Differences by site (n=5) were statistically significant (p< 0.00) with benefits reported by 89. to 98.% of ppts. The percentage of ppts reporting a benefit decreased slightly by age in years (yrs) at enrollment (8-20yrs 229/238=96.2%, 2-25yrs 32/334=93.4%, 26-30yrs 36/46=93.2% and 3-35yrs 75/83=90.4%) but the difference was not statistically significant (p=0.23). Similarly high percentages of both treatment groups, and both sexes reported a benefit at one or more study visits (vaccine 374/400=93.5% vs. placebo 378/40=94.3%; women 339/360=94.2% vs. men 43/44=93.5%). Frequently reported benefits were receiving free HIV risk reduction counselling and testing, and knowing one s HIV status. Twenty male ppts identified information about medical male circumcision was beneficial. Ppts also reported pride in doing something good for [their] community, helping my nation, and educating others about HIV/AIDS. Conclusions: The majority of ppts, including those reporting negative SI events, reported benefits of study participation on their lives. Differences by site were statistically significant (p< 0.00). Additional examination of reported benefits of study participation may be informative, and potentially enhance future recruitment and retention efforts. Impact of DSMB Outcomes on Participation in HIV Prevention Trials: The VOICE Study Experience in Kampala, Uganda Juliane Etima, Teopista Nakyanzi, Samuel Kabwigu, Flavia M. Kiweewa, Brenda G. Mirembe, Carolyne A. Akello, Lisa Rossi 2, Clemensia Nakabiito MU-JHU Research Collaboration, Kampala, Uganda, 2 Microbicide Trials Network, Pittsburgh, PA, United States Background: Clinical trials are monitored and regulated by independent bodies to ensure participants safety and ethical conduct. Depending on study progress, these bodies may recommend a trial to continue as is, be modified or stopped. Six such reviews were conducted during the implementation of the VOICE study. We describe the impact of Data & Safety Monitoring Board (DSMB) recommendations on continued study participation in Kampala, Uganda, including retention, researchers motivation to continue the study, and community perceptions and attitudes about the study. Methods: Communication plans were used to disseminate DSMB outcomes to 6 tiers of stakeholders including study staff, participants, Institutional Review Board, Community Advisory Board, media, and other key stakeholders, including civil society. A qualitative analysis of DSMB outcome reports and review of participant retention was done. Discussion about different scenarios was done in advance with stakeholders. Results: Stakeholders across the 6 tiers expressed disappointment with DSMB outcomes in VOICE. We are ashamed because the doctors treated many infections and we benefited. The tests were expensive and the blood samples showed we did not use the products (participant). All our effort has gone to waste (staff). HIV drugs disappoint researchers (media). We told you that your things will not work, you want to experiment on us (community member). The number of missed visits increased after dissemination of the Nov 20 DSMB outcome when a second study arm (tenofovir gel) was stopped; 24% (4/7) as compared to % (8/7) in Sept 20 when the first arm (tenofovir tablet) was stopped (p< 0.00). Conclusions: Communication about negative DSMB outcomes remains a challenge, although communication plans make dissemination more manageable for sites. Data from VOICE suggest that DSMB outcomes may have had a significant impact on visit retention in Kampala. 364 HIV Research for Prevention 204 HIV R4P

365 Thursday, 30 October Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42.03 Reporting of Adherence in the VOICE Trial: Does Disclosure of Product Non-use Increase at the Termination Visit? Barbara S. Mensch, Ariane van der Straten 2, Elizabeth Brown 3, Karen Liu 3, Jeanne Marrazzo 4, Zvavahera Mike Chirenje 5, Kailazarid Gomez 6, Jeanna Piper 7, Karen Patterson 3 Population Council, New York, NY, United States, 2 RTI International, San Francisco, CA, United States, 3 SCHARP - FHCRC, Seattle, WA, United States, 4 University of Washington, Seattle STD/HIV Prevention Training Center, Seattle, WA, United States, 5 UZ-UCSF Collaborative Research Programme, Harare, Zimbabwe, 6 FHI 360, Durham, NC, United States, 7 National Institute of Allergy and Infectious Diseases, Bethesda, MD, United States P42.04 Reporting of Challenges to Adherence in VOICE: A Comparison of Quantitative and Qualitative Self-reports among Women during and after the Trial Barbara S. Mensch, Ariane van der Straten 2, Miriam Hartmann 2, Helen Cheng 2, Barbara Miller, Jeanna Piper 3, Lisa Levy 4, Cynthia Grossman 5, Elizabeth Montgomery 2, on behalf of the MTN 003D Team Population Council, New York, NY, United States, 2 RTI International, San Francisco, CA, United States, 3 National Institute of Allergy and Infectious Diseases, Division of AIDS, Bethesda, MD, United States, 4 FHI 360, Washington, DC, United States, 5 National Institute of Mental Health, Center for Mental Health Research on AIDS, Bethesda, MD, United States Background: Results from VOICE indicated that 50% of women assigned to active products (Truvada, oral or vaginal tenofovir) had no drug detected in any plasma samples tested during the trial. Yet, in response to questions on product use asked of participants during the trial, 90% of doses were reportedly taken. To explore factors associated with suboptimal adherence, a behavioral exit questionnaire was developed after early closure of the oral tenofovir and vaginal gel arms. The underlying rationale was that women might respond to questions about adherence more candidly at termination than they would during the course of the trial. Methods: For the sub-sample of VOICE participants still enrolled in the trial in December 20, we compared self-reported adherence obtained with the same/similar interviewer-administered questions at monthly/ quarterly visits and at the termination exit visit (TEV) using descriptive statistics and regression models. Results: A TEV questionnaire was administered for 232 of the 5029 VOICE participants. Of the 34 women who reported at the TEV that they always used the product, 53.8% reported missing doses in one of the monthly or quarterly follow-up interviews. Correspondingly, of the 82 women who reported in a monthly/quarterly interview that they always used the product, 34.6% reported in the TEV that they sometimes missed doses. A comparison between a self-rating adherence scale in the quarterly and TEV visits indicated a slightly lower reporting of fair/poor/very poor adherence in the former (7% vs 2%) although the difference was not statistically significant. Conclusions: Participants in the VOICE trial were not more likely to disclose non-adherence at the termination visit than during follow-up visits. While a large proportion of women reporting perfect adherence during the course of the trial reported missing doses at termination, an even larger proportion of women reporting perfect adherence at termination reported missing doses during the monthly/quarterly visits. Background: Results from VOICE revealed that drug was detected in 30% of plasma samples from women assigned to active arms. Since participants indicated 90% of doses were taken, do self-reports on challenges to product use provide useful information on adherence? If so, when, during a trial, is it optimal to collect such information? Methods: We investigate adherence challenges reported during and after VOICE among 72 active arm participants enrolled in MTN-003D, a qualitative study exploring aspects of the trial that affected product use. Responses to questions on challenges to use and reasons for non-use were compared: ) at the first and last VOICE quarterly visits when participants reported on product adherence (PA); 2) at a Termination Visit interview (TV) conducted about 8 weeks after product use ended among 38 of the 72 participants still enrolled when the questionnaire was designed following early closure of two arms; and 3) during the 003D Stage 2 in-depth interviews (IDI), conducted about.5 years after the TV, where participants were informed about their drug detection pattern and classified into adherence groups. Results: At the first PA 6 of 70 participants and, at the last PA, 4 of 68 indicated reasons they did not use product daily in the prior 4 weeks. At TV, 5 of 38 participants responded that they were not always adherent; on average.6 challenges were reported by these 5. During the IDIs after drug results were revealed to participants, 65 of 72 reported challenges; 38 listed 5+ challenges. Among the 43 in the low adherence group, 4 reported no challenges; 26 reported 5+. Among the 20 in the high adherence group, 3 reported no challenges; 9 reported 5+. Conclusions: VOICE participants were reluctant to disclose reasons for non-adherence when on product; after product use ended, willingness to report challenges increased slightly. Only when presented with drug detection patterns denoting whether product was taken, however, were participants forthcoming about the challenges they faced

366 Posters Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42.05 P42.06 Bone Mineral Density Changes among Healthy African Pre-menopausal Women Participating in a Tenofovir-based HIV PrEP Study: The MTN-003B Study Brenda G. Mirembe, Cliff Kelly 2, Mgodi Nyaradzo 3, Suzan Greenspan 4, James Dai 2, Vivian Bragg 5, Jeanna Piper 6, Carolyne A. Akello, Flavia M. Kiweewa, Magure Tsitsi 3, Clemensia Nakabiito, Sharon Riddler 4 Makerere University-Johns Hopkins University Research Collaboration, Kampala, Uganda, 2 SCHARP - FHCRC, Seattle, WA, United States, 3 UZ - UCSF, Harare, Zimbabwe, 4 University of Pittsburgh, Pittsburgh, PA, United States, 5 FHI 360, Durham, NC, United States, 6 DAIDS/NIAID/ NIH, Bethesda, MD, United States Background: Limited data exists on the effect of tenofovir on bone mineral density (BMD) in HIV negative women. This is a major concern for long-term PrEP regimens. We evaluated effects of daily oral tenofovir (TDF) and Truvada (FTC/TDF) compared to placebo on BMD among women in a substudy of the VOICE trial. Methods: HIV negative women in Uganda and Zimbabwe on oral TDF, FTC/TDF, or placebo had BMD measurements of lumbar spine and total hip via dual energy x-ray absorptiometry (DXA) at baseline (BL) and every 24 weeks (w) until 48 w after study product completion. Plasma tenofovir levels were assessed every 2 w for the first 48 w. Student s t-test and regression models were used to compare the mean percentage (%) change from BL in hip and spine BMD between the active and placebo arms. Results: Of 58 women enrolled, 432 (55 Uganda, 277 Zimbabwe) with DXA results at both BL and w 48 entered this analysis. Median age and BMI at BL were 28 years and 24.6 kg/m 2. History of depot medroxyprogesterone acetate (DMPA) use was reported by 23 (28%) and 94% had moderate to high BL physical activity levels. In the primary analysis, there were no significant differences between arms in hip or spine BMD % change, likely due to low product use in VOICE. Tenofovir was detected in 75-00% of plasma samples in 43/8 (36%) TDF and 50/56 (32%) FTC/TDF recipients (median 4 samples tested). In these women, mean % change from BL to w 48 in spine BMD was.0% to.3% lower than placebo and hip BMD was 0.5% to 0.8% lower after adjusting for country, age, BMI, physical activity level, and history of DMPA use (p< 0.05 for FTC/TDF and for oral active arms combined, but not TDF). BMD increased after stopping product; 48 w change was % higher in TDF and FTC/TDF vs placebo recipients. Conclusions: Small but significant reversible decreases in BMD were observed among young African women with higher adherence on TDFbased oral PrEP. The observed differences were in the range seen in prior studies of HIV-negative men and women. Sharing of Investigational Drug among Participants in the VOICE Trial Jeeva Moodley, Leanne Vallabhjee, Sarita Naidoo, Jayajothi Moodley, Gita Ramjee,2 South African Medical Research Council, HIV Prevention Research Unit, Westville, South Africa, 2 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom Background: Randomised controlled trials rely on effective preservation of the random allocation of interventions when tested against placebo for a robust outcome. Product sharing among participants can compromise the trial outcome. In resource poor settings, many participants attend study visits together, travel together and often are neighbours or live in the same household. In such circumstances product sharing poses a real concern. We describe incidents of product sharing across 7 clinical research sites in Durban conducting the VOICE trial and strategies implemented to avert such occurrences. Methods: The Durban sites enrolled 2750 women with 647 and 03 participants randomised to the oral and gel arms respectively. Product use assessments were conducted at monthly visits. Product labels with unique identifiers and product codes were checked to verify return of correct products. Discrepancies prompted discussions with participants and strategies were developed by pharmacists. Monthly product use counselling with product sharing messaging was provided to participants. Results: Thirty two incidents of product sharing were identified: 8 incidents involved participants returning products not assigned to them and 4 incidents included product count discrepancies. Discussions with participants suggested that the main source of product sharing involved women residing in the same household with participants, 20/32 incidents (62.5%). Four participants also reported that visitors had access to their product. Topical gels were more commonly shared than oral tablets (72 % vs. 28 %). Investigations were done to rule out pharmacy dispensing errors. Counselling, tailored messaging and unique identifiers were used to address product use challenges. Conclusions: In this trial, product sharing was common among women residing in the same household. Unique labelling of products helped avert repeat incidents of product sharing. For future trials, targeted counselling and unique product identification is recommended. 366 HIV Research for Prevention 204 HIV R4P

367 Thursday, 30 October Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42.07 An Exploration of Young Women s Perceptions and Experiences of Participating in HV Vaccine Clinical Trials in Nyanga Township, Cape Town P42.08 Racial Differences in Willingness to Participate in HIV Prevention Clinical Trials amongst University Students in KwaZulu-Natal, South Africa Norah Nandudu, Johannes John-Langba,2 University of Cape Town, Department of Social Development, Cape Town, South Africa, 2 The Cairns Institute, Cairns, Australia Diantha Pillay, Douglas R Wassenaar UKZN, Durban, South Africa Background: HIV/AIDS has severely inflicted suffering on the global population and reported to be the worst killer disease in sub-saharan. Because other preventive measures such as condom use among young people is still low and less effective in preventing the spread of the disease. On that basis, it was recommended by the United Nations General Assembly Special Session on HIV/AIDS (200) to accelerate the development of HIV vaccine aimed at curbing the disease. This study sought to explore young women s experiences and perceptions about HIV prevention vaccine clinical trials so as to inform the design and implementation of vaccine trials in Africa. Methods: The study employed purposive sampling to select participants and qualitative research design was used to interview participants using semi-structured interview schedule. A tape recorder was used to capture data and coding procedures were used to analyze data. Results: Findings drawn from participants responses and compared with literature from previous studies on vaccine trials and social development theories indicate that participants decide to join HIV prevention vaccine clinical trials because they hope to be protected from HIV infection. Most importantly participants hope to get access to medical care and treatment, meanwhile some participants perceived HIV vaccines harmful to humans hence they usually decline to participate. The study also identified study participation challenges related to socio-cultural and historical aspects. Conclusions: Although vaccines have had some success stories in the prevention and control of infectious diseases such as the eradication of polio, smallpox and measles, prevailing challenges need to be addressed if vaccine development is to be feasible. Providing more information, reinforcement of community awareness and mobilization around issues of HIV vaccine clinical trials at all levels of vaccine design and implementation is required to ensure appropriateness and acceptability of vaccine trial participation. Background: Willingness to participate in clinical trials is a crucial element in recruitment of suitable participants for intervention trials. Measuring willingness to participate helps determine community preparedness for clinical trials. Researchers in the USA developed a Clinical Research Involvement Scale (CRIS) assessing willingness to participate modelled on the Theory of Reasoned Action. Methods: This study aimed to determine racial differences in willingness to participate and explore potential factors associated with willingness to participate in HIV prevention research using the CRIS. The CRIS was administered online with a demographic questionnaire to university students aged 8-45 at the University of KwaZulu-Natal, South Africa. Associations between willingness to participate and age, gender, relationship status, parity, religion, education status, student status, employment status and access to private health care were examined. Results: The study enrolled 636 participants, two thirds being female. After data cleaning a sample of 509 was considered for analysis. Results indicated that all students across all race groups were willing to participate in HIV prevention research. However, there was a statistically significant difference in factors affecting willingness of participate. Based on the differences amongst these factors, Black students expressed greater intention to participate compared to White and Indian students. Racial differences in factors that affect willingness to participate indicate differences in risk perception and seeking access to better quality healthcare. Conclusions: The CRIS is a reliable instrument in this population; however in its current structure it does not show strong validity. Validity improved if factors of motivation to comply and outcome evaluations were removed. The CRIS should be used in other populations to assess its validity

368 Posters Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42.09 P42.0 Use of Visual Education Aids to Improve Participant Understanding of HIV Prevention Trials Nonzwakazi Mnqonywa, Renee Street Medical Research Council, HIV Prevention Unit, Westville, South Africa Background: The HIV Prevention Research Unit (HPRU) based in KwaZulu-Natal (South Africa) has been involved in numerous HIV clinical trials and has tested a range of products including pills, gels, diagrams and rings. Over the years, visual aids have been created to improve participant understanding of the study product, design and procedures Methods: The use of educational materials such as flip charts and booklets were developed to go hand in hand with visual aids to overcome potential language barriers as well as to compensate for level of education when explaining various aspects of the clinical trials. Visual aids are designed by Family Health International in conjunction with input from clinical trial research staff before they are sent for ethical approval. Results: The study population typically consists of sexually active females between the ages of The socio-cultural, economic & demographic determinants of the participants vary and level of education ranges from illiterate to highly educated. The majority of participants have a clearer understanding of the purpose of the study after using visual aids. Through the years, visual aids have evolved from pictures to real life models. These include sample applicators, vaginal rings, dildos to demonstrate condom use, pelvic model and other different types of family planning methods. Some studies use an audiocomputer-assisted-self interview system (ACASI) which includes visuals to make understanding and operating more user friendly. Conclusions: Illiteracy is no longer an excuse for HIV prevention education. HIV clinical trial participants, their male partners and the communities at large have demonstrated a clear understanding of study products, designs and procedures with the use of visual aids. Preparing for HIV Prevention Trials: From Training to Service Delivery-lessons Learned with Long Acting Reversible Contraception (LARC) in Zambia Mubiana Inambao, Naeemah Munir, Kathleen Wu 2, Danielle Dang 2, Nurilign Ahmed 3, Bethelihem Getachew 4, William Kilembe 2, Bellington Vwalika 2, Amanda Tichacek 5, Susan Allen 5 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Ndola, Zambia, 2 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Rwanda Zambia HIV Research Group, Projet San Francisco, Kigali, Rwanda, 4 Rwanda Zambia HIV Research Group, Emory University, School of Medicine, Atlanta, GA, United States, 5 Rwanda Zambia HIV Research Group, Emory University, Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA, United States Background: Zambia has a high burden of mother-to-child transmission of HIV. The provision of long-acting reversible contraceptives (LARC), specifically intra-uterine devices (IUDs) and Jadelle implants, is critical to prevent unplanned pregnancy and perinatal transmission in HIV+ women and is also necessary to prevent pregnancy in HIV- prevention trial participants. The UK Department for International Development has funded a program to avert an estimated 3,900 new HIV infections through integrated PMTCT with LARC and HIV couples-focused HIV testing. Methods: LARC training for providers began with a 3-day didactic training followed by supervised IUD and Jadelle insertion and removal practicums. Those showing proficiency in all 4 categories were considered fully certified. Practicums were conducted at government clinics in Copperbelt, Lusaka and Southern Province. Results: From June 203 to March 204, ZEHRP trained 99 LARC providers from 45clinics. 9% were certified in Jadelle insertions, 70% in Jadelle removals, 55% in IUD insertions, 46% in IUD removals and 43% were fully certified. The main obstacles to completion of practicums were not being assigned to the family planning department when one of the few trainers was available, and coordinating these times with availability of interested LARC clients. A Training-of-Trainers was held for the fully certified nurses who were high performers, meetings were held with clinic in-charges to coordinate trainee schedules, and satisfied clients were recruited to give personal testimonials. Conclusions: IUD and implants are rarely used in Africa, primarily because lack of supply leads to lack of demand. This model illustrates an effective way to harness successful LARC trainees as trainers and work with clinic administrators and satisfied clients to coordinate practicum training and demand creation. Repeating this cycle leads to a snowball effect of progressive and simultaneous increase in supply and demand, resulting in PMTCT and trial-ready clients with LARC. 368 HIV Research for Prevention 204 HIV R4P

369 Thursday, 30 October Posters 42: Participation in Trials: Willingness, Benefits and Challenges P42. Implementation of an Adherence Counseling Intervention in a Microbicide Trial: Challenges in Changing Counselor Behavior Iván Balán, Alex Carballo-Diéguez, Rebecca Giguere 2, Javier Lama 3, Ross Cranston 4 NYSPI/Columbia University, HIV Center for Clinical and Behavioral Studies, New York, NY, United States, 2 New York State Psychiatric Institute, HIV Center for Clinical and Behavioral Studies, New York, NY, United States, 3 Asociacion Civil Impacta Salud y Educacion, Lima, Peru, 4 University of Pittsburgh Medical Center, Division of Infectious Diseases, Pittsburgh, PA, United States Background: Adherence counseling interventions have become a critical component of biomedical HIV prevention trials. Yet, little is known about how well counselors learn and deliver them. We present interim findings about implementing the participant-centered adherence counseling intervention being used in MTN-07. Methods: Counselors at seven study sites completed two days of training on delivering the standardized intervention. Next, they conducted two practice sessions, at least one of which had to meet pre-determined fidelity criteria for approval to see study participants. All sessions were audio-recorded. For each counselor, the practice sessions, the first ten study sessions, and one of every five subsequent sessions were rated for fidelity. Ratings were used to guide monthly group coaching sessions with the counselors. Results: Twenty-six counselors completed the two-day training. For 5 counselors (58%), both practice sessions met fidelity criteria; for seven counselors (27%), one session met criteria; and for four counselors (5%), neither did. To date, 226 of 227 counseling sessions conducted have been recorded. Of the eight counselors who have completed ten study sessions, six met fidelity criteria on 80% or more of their sessions, but two met criteria on 50% or less of the sessions. Fidelity ratings varied over time. Fidelity ratings of the individual components of the sessions suggest that lower ratings were mostly attributable to skipping components of the session, not to conducting them incorrectly. Conclusions: Recording adherence counseling sessions during biomedical trials is feasible and acceptable to counselors and participants. After 2 days of training, the majority of counselors, though not all, were able to accurately deliver the intervention. Over time, counselors appear vulnerable to lapses in fidelity. Findings highlight the need to assess efficacy of training for such interventions and for continued monitoring and coaching throughout the study to ensure fidelity

370 Posters Posters 43: Policy, Advocacy and Modeling P43.0 P43.02 Should the United States Policy that Banned MSM from Donating Blood be Repeal? Olanrewaju Adedokun Southern Connecticut State University, Public Health, New Haven, CT, United States Background: Recently, the American Red Cross, which is the largest single supplier of blood and blood products in the United States, issued an emergency request for donors of all blood types because its blood donations were down by about 0 percent across the country, with about 50,000 fewer donations than expected. This led to the question of whether the United States policy that banned men who have had sex with other men (MSM), at any time since 977 from donating blood be repealed to allow gay and bisexual men with no HIV infection, who meet the other conditions and are willing to donate blood be allowed to do so. That permitting blood donation from the eligible MSM could help reduce blood supply shortage in the United States. However, the initiator of that policy, U.S. Food and Drug Administration maintained that the life time ban will not be eased unless there is an evidence-based data showing that a repeal of existing policy is necessary and will not put the life of blood recipients at a significant risk of acquiring HIV infection through blood transmission. Methods: This study analyze the views of the interested stakeholders of this issue by weighing the evidences presented by those who want the FDA s current policy to be repealed with the evidences of those who does not. Results: Having reviewed all the parties views in this case, three options were identified and they are as follows: Not repeal the FDA s current policy (life time ban); Repeal the FDA s current policy (life time ban) and replace it with one- year deferral; and Repeal the FDA s current policy (life time ban) without deferral option. Conclusions: Having carefully reviewed all the available evidences, the best option would be to repeal the current policy (life time ban) and replace it with one- year deferral. This option addresses the concerns of every party involved. It safeguards the life of patients who received donated blood from HIV infection and removed life time ban on the gay community to one year deferral. Streamlining Operational Processes by Building a Centralized HIV Clinical Research Centre Jacqueline S. Burgess, Krishnaveni Reddy, Pranitha Ramchuran, Clare Dott, Thesla Palanee, Helen Rees Wits Reproductive Health & HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: Wits RHI is situated in Hillbrow, Johannesburg, a model community to conduct HIV clinical studies, with its high prevalence rates of HIV and developing infrastructure. Wits RHI has created a multifunctional Research Centre (RC) where 2 clinical and observational research studies have been conducted over the past 4 years. Given the financial constraints on investigator driven research and clinical trials, the RC has formulated centralized operational services that support all ongoing studies. Methods: The centralized operations of the RC involves the sharing of data, laboratory, pharmaceutical, regulatory, administration and maintenance capacities across all studies. This includes the sharing of fixed costs in terms of infrastructure, licensing, and equipment; offering of voluntary counselling and testing; provision of concomitant medication; shared data, administration, and laboratory centers; cross-trained staff for back-up purposes; and support from centralized maintenance and operational teams. The RC uses the MRC Biometric Co-Enrolment Prevention System, which detects if participants are enrolled in other studies, to eliminate the possibility of co-enrolments. The Division of AIDS standards is used as a benchmark for quality across all studies. Management decisions are aimed at balancing the needs of individual studies and of the site. Results: Centralizing services improves cost efficiencies for all studies, while providing access to optimized laboratory, data, regulatory support, clinical staff and equipment, pharmaceutical services, site infrastructure and staff capacity. This approach has resulted in improved quality of the site s study implementation and overall outputs and has reduced duplication of efforts in the organization. Conclusions: With the current limited funding available for HIV clinical research, developing centralized processes for research sites is a potential mechanism for reducing costs and optimizing outputs and of benefit for sites with financial limitations. 370 HIV Research for Prevention 204 HIV R4P

371 Posters 43: Policy, Advocacy and Modeling P43.03 Thursday, 30 October P43.04 Policy Dialogue to Enhance the HIV Response among MSM and Transwomen in Peru through Combined Prevention Strategies: A Baseline Stakeholder Assessment Carlos F. Caceres, Ximena Salazar, Alfonso Silva-Santisteban, Aron Nunez-Curto Cayetano Heredia University, Unit of Health, Sexuality and Human Development, Lima, Peru Background: Despite significant changes in the HIV epidemic landscape among MSM and transwomen (TW) to date, the national program remains essentially as it was designed in the late 990s, with limited discussion of potential changes. Together with the Ministry of Health, we are conducting, documenting and analyzing a process seeking to enhance the program with combined prevention strategies, through stakeholder involvement, health systems assessments and mathematical modelling. Here we present the background stakeholder analysis. Methods: We conducted in-depth interviews and focus groups with community members (MSM/TW), in-depth interviews with community leaders, health providers and decision makers, and two policy discussion workshops (with civil society and government representatives, respectively). Results: Among community members and stakeholders alike, condom use is still the epitome of prevention, and its limited use is explained only as resulting from poor individual commitment and insufficient prevention work. Knowledge of ETfP is almost absent, but there is some knowledge of PrEP, together with resistance to its potential use. The lack of information, misconceptions and confusion reflect limited public discussion on new HIV prevention technologies. On an operational level, participants consider the need of well-trained health providers as part of a renovated program. People agreed with the idea of comibined biomedical, behavioral and structural strategies based on appropriate evidence. All recognized the need for continuing professional HIV educaiton, and also for spaces to evaluate and improve current programming. Renovated leadership will play a key role. Conclusions: Community members and stakeholders in Peru agree on the need to bring the HIV prevention response to date among MSM/TW with combined strategies. Many feel that HIV prevention programming is loosing momentum and needs new leadership and tools, and are interested in an evidence-based process where the HIV response can be reconceptualized. Forging Consensus in the HIV-positive Community on the Use of Treatment as Prevention Gus Cairns,2, Brian West 2,3, Caspar Thomson 4, A Cornelius Baker 5, Anna Zakowicz 6,7, Nikos Dedes 8 NAM Publications, Editor, London, United Kingdom, 2 European AIDS Treatment Group, Brussels, Belgium, 3 Waverley Care, Edinburgh, United Kingdom, 4 NAM Publications, London, United Kingdom, 5 FHI 360, Washington, DC, United States, 6 Global Network of People Living with HIV (GNP+), Amsterdam, Netherlands, 7 AIDS Healthcare Foundation, Amsterdam, Netherlands, 8 Positive Voice, Athens, Greece Background: HIV treatment policy has been moving closer towards access to ART for all. This has not been universally welcomed among HIV advocates. Concerns have centred around whether TasP could lead to compulsory and oppressive test-and-treat (T&T) policies and whether TasP is the best use of resources when many needing ART for life still do not get it. These exist alongside enthusiasm for TasP and determination that disadvantaged populations should benefit from it. Such tension in the community has led to the lack of a simple, strong community-led policy on TasP. Methods: EATG ( and NAM ( devised a statement of broad principles for any TasP programme. This sign-on statement is a guide to ensure that all ART programmes centre on the interests of people with HIV. Devising the statement was an exercise in forging community consensus. The statement went through consultation with the EATG membership, a public consultation on an international community meeting and final consultation with key opinion leaders. It was published in February 204 on Results: Feedback during consultation said that the statement was too technical and its authorship and intended audience unclear; it needed to be sited clearly within a human rights background; there should be more mention of gender and power issues; and it needed to support existing effective prevention methods. Sections on the evidence for TasP and research needs became separate appendices. The statement now consists of the main sign-on statement of 30 clauses, an -page fully-referenced statement including the appendices and a Key Points summary. Conclusions: The statement was presented at the 20 TasP Workshop and in its own session at the International AIDS Conference and of an article in Communicable Infectious Diseases. We aim to have the statement incorporated into future guidelines and to work with partners to ensure that TasP is firmly grounded in the rights of people living with HIV. 37

372 Posters Posters 43: Policy, Advocacy and Modeling P43.05 Do Civil Society Advocates Priorities Align with HIV Prevention Research Landscape? Lisa Jacobs, Manju Chatani, Angelo Kaggwa-Katumba AVAC, New York, NY, United States P43.06 Making Treatment as Prevention Work for People with Disabilities across the HIV Prevention and Care Continuum in Zambia Clever Chilende Treatment Advocacy and Literacy Campaign (TALC), Programmes, Lusaka, Zambia Background: HIV prevention research-to-rollout processes require effective community advocates who understand the evolving landscape. The AVAC Advocacy Fellowship, launched in 2009, supports advocates to engage around these issues in their countries. AVAC sought to understand how applicants priorities relate to developments in the field. Methods: Desk analysis was conducted on 284 eligible applications of over 400 submitted in the five years of the program to determine which interventions were of greatest interest each year and if any patterns emerged related to events in the field. Results: Voluntary medical male circumcision (VMMC) was identified in 9 applications, 80 of them received from Interest waned as countries ramped up, and PEPFAR began funding, VMMC programs. Microbicides were identified in 58 applications; however, 23 applications were received in 2009 and only 9 applications in after CAPRISA 004 results were announced and there was confusion about what happens next. PrEP was identified in 57 applications with a surge in 202 after the FDA approved Truvada for PrEP. Treatment as prevention was identified in 4 applications, all of which were received in after HPTN 052 results were announced. While vaccine research has received relatively little interest, it was highest in 2009 just after RV44 showed positive results. Conclusions: Applicants interests seem to follow the events in the field, demonstrating that advocates are aware of and interested to localize global events. Since AVAC amends its application materials every year and highlights priorities on its website, fluctuations in choice of interventions may reflect a selection bias. However, the trends are dramatic and reflect the evolving issues and interests of applicants to the Fellows program. Since AVAC can only offer a few Fellowships each year, more efforts are needed to support the large and informed pool of HIV prevention research advocates who are interested in engaging in the field. Background: Since 2005, Zambia has developed a national universal HIV prevention and testing program with free antiretroviral. Although prior research in sub-saharan Africa has shown that people with disabilities are highly vulnerable to HIV, little research on this population has been done in Zambia, and the national HIV program has only recently included people with disabilities as a vulnerable population in its national strategy. Methods: In-depth interviews with adults, children and adolescents with physical, sensory and psychosocial disabilities were conducted in Zambia. Caregivers and parents of children with intellectual disabilities were also interviewed, as were health workers, special education teachers and representatives of disabled persons organizations and HIV organizations. Legal and policy analysis was also conducted. Results: Persons with disabilities experience pervasive stigma and social marginalization compounded by economic, communication and physical barriers to access to HIV services throughout the HIV care continuum in Zambia. HIV prevention education and standard protocols for assuring informed consent and confidentiality in the provision of HIV testing and encouraging ART adherence need to be tailored to the specific needs of individuals with disabilities. Existing HIV and health policies do not identify specific strategies and interventions to address people with disabilities. The equal right of people with disabilities to sexual and reproductive health is not recognized and they remain invisible to social protection services. Conclusions: Efforts to achieve universal access to HIV prevention and treatment in Zambia are undermined by the lack of programs addressing the needs of individuals with disabilities, and current policies and programs on HIV and disability lack adequate integration. Recognition of people with disabilities as a highly vulnerable group within the national HIV response has not been translated into implementation of inclusive and targeted services. 372 HIV Research for Prevention 204 HIV R4P

373 Posters 43: Policy, Advocacy and Modeling P43.07 Not without us: Mobilizing Communities to Advocate for Women s Involvement in PrEP Implementation Thursday, 30 October P43.08 Use of a Mathematical Model to Predict Dissolution Profiles of Dapivirine Vaginal Rings Dazon D. Diallo SisterLove, Inc., Atlanta, GA, United States Christopher Gilmour, Stephen Ampofo, Brid Devlin International Partnership for Microbicides, Product Development, Silver Spring, MD, United States Background: As the US FDA considered the approval of the use of Truvada as PrEP, women in the US organized to address the need for greater community engagement in the implementation science informing the introduction of the intervention in clinics, community health centers and private care.through an ad hoc coalition, US women s communities advocating for safe HIV prevention options for women, including PrEP, convened a strategic working group dedicated to providing input, community-based qualitative research, and innovative strategies research to determine how to help women and their healthcare providers make PrEP a viable option. Methods: Community engagement in research includes activities beyond the involvement of volunteers in trials, community advisory boards and local partnerships with service organizations.our methodology included a diversity of initiatives including: informational webinars/community symposia, key informant interviews, qualitative study with focus groups and surveys; implementation advocacy; engaging federal partners. Results: The work resulted in the following: convened 3 national/ international webinars for a total of 850 participants; captured qualitative data on knowledge, attitudes and beliefs of 85 women at high risk for transmission; published several articles/op-eds in newsprint; presented papers, posters and workshops at national and international meetings; and sustains a working group of nearly 80 interested parties/organizations. Conclusions: The implementation of research, such as PrEP, requires meaningful involvement and representation from the community most affected by the research outcomes.in the US, and key countries where PrEP is a priority, women s education, understanding and acceptance of PrEP is an important step to its use as a prevention option.the community mobilization of women s health/hiv advocates, clinical and behavioral/ social scientists & WLwH is a key component of engaging community in research of a product before and after it has been approved. Background: The dapivirine vaginal ring is a sustained release formulation containing dapivirine, a potent antiviral currently in clinical evaluation for HIV prevention. Each ring contains 25 mg of dapivirine in a platinum-cured silicone matrix. In vitro dissolution testing quantifies the amount of dapivirine released from the ring over the 28-day use period, but is labor intensive, time consuming and expensive. This paper describes development of a mathematical model to predict average cumulative drug release of a batch. Methods: Empirical approaches and mechanistic considerations were used to modify the Higuchi equation so as to assess the amount of dapivirine released per unit area of the vaginal ring. The model utilizes details of ring geometry, ring assay and a single attribute of the silicone elastomer: Q = (2A) /2 (C p *D p ) /2 X {polynomial (order 2) of t} Where Q = amount of active ingredient released per unit area A = concentration of active ingredient in the matrix Cp = solubility of active ingredient in the matrix Dp = diffusion coefficient of the active ingredient in the matrix t = time, set at 28 /2 The model was assessed for applicability to batch averages. Average 4- day results were also used to estimate (C p *D p ) /2 for individual batches in order to improve the model. Results: The model resulted in errors typically within the range +/- 4% for the predicted 28-day average cumulative release versus actual average cumulative release. The accuracy of the model was improved further by using the 4-day results to estimate (C p *D p ) /2 for individual batches. Conclusions: Data from mathematical models indicate that ring attributes or results from earlier time points can predict 28-day dissolution in terms of average cumulative amount of drug released per batch or from an individual ring. These predictions provide a reliable and efficient measure for product quality control

374 Posters Posters 43: Policy, Advocacy and Modeling P43.09 Trends of Advocacy Journalism; a Case of the HIV/AIDS Story in the Ugandan Press Kakaire A. Kirunda,2, Angelo Kaggwa- Katumba 3 Islamic University in Uganda, Mass Communication, Kampala, Uganda, 2 Makerere University College of Health Sciences, School of Public Health, Health Policy Planning and Management, Kampala, Uganda, 3 AVAC: Global Advocacy for HIV Prevention, New York, NY, United States P43.0 Tenofovir Diphosphate Concentrations in Human Vaginal Stroma after Different Dosage Regimens with a Vaginal Gel: A Modeling Approach Yajing Gao, Andrew Yuan, David F. Katz,2 Duke University, Durham, NC, United States, 2 Duke University Medical Center, Obstetrics and Gynecology, Durham, NC, United States Background: The genre of advocacy journalism is one which journalists can employ to advance HIV prevention efforts ranging from policy to practice. Advocacy journalism deliberately but transparently adopts a non-objective viewpoint, usually for some social purpose. Usually, in advocacy journalism, practitioners have an opinion about the story they are writing. We therefore sought to establish the extent to which the strategically placed front page HIV/AIDS stories in Uganda s leading daily newspapers are a product of advocacy journalism. Methods: A retrospective desk review of the Daily Monitor and New Vision newspapers covering the period January 203 through December 203 was used. The two newspapers were purposively selected and HIV/AIDS articles obtained from the cover pages were coded against a set of variables. Two key informants were also interviewed. Data was entered into SPSS (Statistical Package for the Social Sciences) software for analysis while qualitative data from the articles was analysed using thematic content analysis. Results: The sample for both newspapers yielded a combined 254 articles on the front pages. However, findings indicate that out of these, only 28 articles (.3%) were about HIV/AIDS with the Daily Monitor having 6 while New Vision carried only 2. Overall, most of the articles had an element of advocacy journalism. In the New Vision, 9 (75%) front page articles were slanted towards advocacy, while in the Daily Monitor this was exhibited in 4 (88%) articles. In both newspapers, the journalists systematically employed a positive tone, the choice of facts supported the cause the headlines were fronting and sourcing for the articles was favourable. All these attributes are synonymous with advocacy journalism. Conclusions: With majority of HIV/AIDS articles that make it to the front pages being products of advocacy journalism, prevention advocates need seize the opportunity. Ways of working with the authors of these articles should be devised to tap into advocacy opportunities. Background: Different vaginal dosing (BAT24, daily) of the % Tenofovir (TFV) gel gave differing pharmacokinetics and prophylactic efficacy. The kinetics of stromal Tenofovir diphosphate (TFV-DP) production/loss are governed by many factors, e.g.: mass transport kinetics of TFV to stroma; concentration distribution of stromal host cells; TFV binding and phosphorylation rates to/in host cells; and TFV-DP clearance rate in host cells. Experimental PK studies give guidance on creation and persistence of stromal TFV-DP levels, but optimization of dosing to achieve such levels is limited. Modeling this process complements experimental studies, providing insights on how the multiple factors govern TFV-DP levels and suggesting optimal dosing strategies. Methods: A mechanistic, mass transport-based model was created of TFV delivery to human vaginal mucosa by a spreading gel, and coupled TFV-DP production in stromal host cells. Parameters in the model were obtained from in vitro measurements of gel rheology and TFV transport properties, human vaginal morphometric and histological data, and results of human PK studies for the % TFV gel. Single and multiple gel application regimens were studied, including coitus. Results: Results show, for example, that application of two doses 4 hours apart (as found in BAT24) gives 40% higher maximum stromal TFV-DP concentration than daily dosing. This elevated TFV-DP concentration is sustained for 4 days vs daily dosing (due primarily to the long half life of TFV-DP). Conclusions: Modeling provides additional insights about interactive effects of the multiple factors that govern optimal dosing by the TFV gel. Results here illustrate pharmacokinetic benefits of multiple dosing within a few hours vs sustained daily dosing. This approach can be used to help optimize dosage regimen, accounting for factors such as gel volume and drug loading. 374 HIV Research for Prevention 204 HIV R4P

375 Posters 43: Policy, Advocacy and Modeling P43. Thursday, 30 October P43.2 Persistent HIV- related Stigma in Rural Uganda during a Period of Increasing HIV Incidence Brian T. Chan,2, Sheri D. Weiser 3, Yap Boum 4, Mark J. Siedner, A R. Mocello 3, Jessica E. Haberer, Peter W. Hunt 3, Jeffrey N. Martin 3, Kenneth H. Mayer 5, David R. Bangsberg, Alexander C. Tsai Massachusetts General Hospital, Boston, MA, United States, 2 Brigham and Women s Hospital, Boston, MA, United States, 3 UCSF, San Francisco, CA, United States, 4 Epicentre, Mbarara, Uganda, 5 Fenway Health, Boston, MA, United States Background: Uganda is the only country in sub-saharan Africa with increasing HIV incidence during the period Because HIVrelated stigma is associated with reduced uptake of HIV testing, increased risk-taking behavior, decreased adherence to anti-retroviral therapy (ART), and reduced HIV status disclosure, it is critical to understand how changes in HIV incidence have correlated with HIV-related stigma during this period. Methods: We analyzed data from two sources: ) the Uganda AIDS Rural Treatment Outcomes (UARTO) study during and 2) the Uganda Demographic and Health Surveys (DHS) from 2006 and 20 to estimate trends in internalized stigma among people living with HIV (PLHIV) at ART initiation and trends in stigmatizing attitudes and anticipated stigma among the general population. We fit regression models adjusted for socio-demographic characteristics, with year of cohort entry/dhs as the primary explanatory variable. Results: Among people initiating ART in the UARTO study, we found a statistically significant positive association between year of ART initiation and internalized stigma score (adjusted b=0.7; 95% CI, 0.05 to 0.29), suggesting an % relative increase in the mean score in each year of recruitment after the first year. From.4 (out of 4) at baseline, the mean stigma score increased to a peak of 2.2 in 20. Among the general population comparing 20 with 2006 DHS data, we found a significantly higher odds of reporting anticipated stigma (adjusted OR =.2; 95% CI,.08 to.7), despite a decreased odds of reporting stigmatizing attitudes (adjusted OR = 0.90; 95% CI, 0.87 to 0.93). Conclusions: Mean internalized stigma at ART initiation increased over time among PLHIV in a rural Ugandan cohort in the setting of worsening anticipated stigma among the general population. Further study is needed to better understand the reasons for increasing HIV-related stigma and its impact on HIV prevention efforts in Uganda. Using Advocacy, Guideline and Policy Change to Increase Treatment Demand in Various Parts of Africa and Asia Maureen Akumu Milanga, Ntando Yola 2, Oladayo Taiwo Oyelakin 3, Peter Michira 4, Rumbidzai Mapfumo 5, Josephine Kamarebe 6, Cai Lingping 7 AIDS Law Project, Nairobi, Kenya, 2 Networking HIV/AIDS Community of South Africa, Cape Town, South Africa, 3 Positive Action for Treatment Access, Lagos, Nigeria, 4 Partners in Prevention PrEP Thika Study Site, Thika, Kenya, 5 Centre for Sexual Health and HIV/AIDS Research, Harare, Zimbabwe, 6 Health Development Initiative, Kigali, Rwanda, 7 China HIV/AIDS Information Network, Beijing, China Background: Through the AVAC fellowship, selected advocates from Kenya, Rwanda, Nigeria, Zimbabwe, South Africa and China worked with the aim of empowering the population with knowledge and to advocate for treatment as prevention in their countries that targeted the community and policy makers. Methods: Advocacy took place in the span of a year where three advocates from Kenya, Rwanda and China worked to increase the demand and access of PrEP in their countries. They targeted key populations consisting of sex workers, gay men, male sex workers and sero-discordant couples, policy makers and civil society. In the communities they created awareness for the program among the recipients and other non-governmental organizations and advocated for government to adopt the program for the country. In Zimbabwe, South Africa, Nigeria and Kenya, four advocates increased demand and awareness of treatment as prevention and good participatory practice in clinical trials. The Kenya, Zimbabwe and Nigeria advocates increased demand by advocating for early initiation of people living with HIV, initiation of sero-discordant couples regardless of CD4 count in Kenya, increase of post exposure prophylaxis access to sex workers in Zimbabwe and in South Africa increase of effective engagement of communities in trials through the development of treatment guidelines and change in policy in Kenya, Zimbabwe and Nigeria and engaging researchers and communities is south Africa. Results: There is need for scale up of PrEP programs in countries with key populations as consultations in the targeted countries showed a demand for the services. There is need to create more community engagement in programs to also increase access among populations. Early initiation and lifelong treatment among mothers is timely and the demand among communities is also high, strides should be taken to fully roll out these programs. Conclusions: Civil society groups in other countries should also seek to create demand among communities on new prevention technologies

376 Posters Posters 43: Policy, Advocacy and Modeling P43.3 Ending HIV among Men who Have Sex with Men (MSM) in Kenya: Community Recommendations on Scaling up HIV Prevention and Treatment Jeffrey W. Wambaya, Leonard Mutisya 2, Gaudensia Mutua 3, Jack Ndegwa 4, George O. Owino 5 Ishtar MSM, Nairobi, Kenya, 2 UHAI EASHRI, Nairobi, Kenya, 3 KAVI, Nairobi, Kenya, 4 KANCO, Nairobi, Kenya, 5 IAVI, Nairobi, Kenya P43.4 Using Advocacy for the V condom - A New Female Condom Coming to South Africa - Building Support for Female Condoms as an MPT Marion Lynn Stevens,2, Penny Parenzee, Jane Bennett 2, Kimberly Whipkey 3 WISH Associates, Cape Town, South Africa, 2 African Gender Institute, University of Cape, Cape Town, South Africa, 3 PATH, Washington, DC, United States Background: MSM in Kenya contribute 5% of new HIV infections yearly even when only % of men report same-sex sexual activity. There has been no conclusive participatory strategy that has engaged MSM nationally in developing HIV prevention responses for and by MSM. Kenyan MSM community organisations purposed to design and advocate for a combination package to advise planning, coordination and evaluation of HIV prevention, treatment, research and advocacy and development of the Kenya National AIDS Strategic Plan IV Methods: Between August and December 203 with leadership of NACC and NASCOP and support from Ishtar MSM, IAVI and LVCT Health, participatory community discussions with 27 MSM-serving organizations designed a package of approaches to ending HIV among MSM. Results: There was consensus that a combination package for MSM be defined and guidelines written. The package must prioritize strategies to improve outreach and access to HIV testing, adopting a find-testtreat-retain model to immediately link HIV-infected MSM to treatment regardless of CD4 count. Additionally, MSM be provided a broad range of prevention options including scaling up access to condoms and condom-safe lubricants, STI testing and treatment, post-rape care, PrEP and PEP. Legal reforms and democracy are critical to fighting stigma and discrimination in the public and among service providers. We must also increasingly and meaningfully engage MSM in critical decision-making in service delivery and research design and implementation, including on PrEP, vaccines and microbicides. Conclusions: Funding, research and advocacy strategies informed by community insight should be prioritized to make HIV prevention investments efficient and sustainable. MSM-led health organizations should be supported to better sustain their work and to integrate and scale up biomedical interventions and research in their existing package of behavioural and structural interventions because of their community familiarity and grassroots outreach strategies. Background: South Africa (SA) launched a new contraception policy in 204 that highlights linkages between HIV prevention and sexual and reproductive health (SRH). HIV-prevention campaigns in SA have focused primarily on male condoms, while female condoms the only available woman-initiated multipurpose prevention technology (MPT) have not been strongly supported. Research suggests that knowledge about female condoms is limited and myths and mistrust persist, which contributes to limited access. The V condom (developed by PATH as the Woman s Condom) is being evaluated for early market introduction in SA. Methods: WISH Associates, AGI/UCT, and PATH collaborated on activities to strengthen the policy and advocacy environment for female condoms overall, and for the V condom in particular. We conducted a policy/advocacy analysis in SA, and identified opportunities for strengthening female condom programming. From , we implemented strategic activities engaging policymakers, health care workers, journalists/media experts, researchers, SRH advocates, and potential consumer groups. These activities raised awareness and generated interest in female condoms overall, and the new V condom specifically. Results: A range of activities strengthened the environment for female condom promotion and awareness, including: developing a cadre of female condom ambassadors who promote female condoms among diverse communities, organizing public awareness days, crafting media guidelines to strengthen reporting on female condoms, and a compilation of digital stories that describe why female condoms are important to women and men in South Africa. Conclusions: Advocacy partnerships and interest from decisionmakers for female condom programming suggest that support for female condoms has gleaned media and public interest. Advocacy helped pave the way for V condom introduction and offers new hope in a country where women have limited options for protecting their reproductive health. 376 HIV Research for Prevention 204 HIV R4P

377 Posters 43: Policy, Advocacy and Modeling Thursday, 30 October P43.5 LB Modeling the Impact of Targeting Treatment and Prevention to the Migrant Population of Male Miners in a Generalized Epidemic Setting Daniel J. Klein, Anna Bershteyn, Kevin Oishi, Philip Eckhoff Institute for Disease Modeling, Bellevue, WA, United States Background: Migrant populations are thought to have played a significant role in the initial spread of HIV in sub-saharan Africa. Miners in particular have been identified as a key driver of the HIV epidemic in South Africa due to their circular migration patterns and elevated risk behaviors. We used a computer model to quantify the potential impact of targeting treatment and prevention to male miners. Methods: We augmented an individual-based network model, EMOD- HIV v0.8, to include a migrant population representative of South African miners. Simulated miners routinely migrate between their home community and a mining community, where an external incidence source replaces sexual transmission. Results: Targeting miners with treatment, perfect prevention, or combination treatment and prevention averted 2.7, 203.3, and 29.8 thousand infections over a 20-year period with 3% discounting, respectively. These results come from baseline assumptions that 5% of the male population engages in mining and that each miner visits home monthly for one week. In comparison, universal test and treat resulted in 3.8 million discounted infections averted over the same time period. We increased the frequency of home visits, however the impact changed only marginally. We then exaggerated the size of the male mining population to 20%, resulting in 62.5, 890.6, and thousand discounted infections averted from treatment, prevention, and combination intervention. Finally, we changed the timing of the prevention intervention from 205 back to 985 in -year increments. To halve the incidence rate in 2020, prevention needed to start in 989. Result may underestimate impact by not modeling transmission at the mine. Conclusions: While targeting miners can be cost effective, here we see that these interventions will have a modest impact on the overall incidence in a generalized epidemic. Nonetheless, targeting migrants remains a promising tool for mitigating the future burden of HIV in lowlevel and concentrated epidemics

378 Posters Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44.0 P44.02 N-alkyl/aryl-4-(2-Substituted-3- Phenylpropyl) Piperazine--Carbothioamide as Vaginal Microbicide with RT Inhibition: Synthesis, Docking and PK Studies Veenu Bala, Bhavana Kushwaha 2, Yashpal S. Chhonker 3, Shagun Krishna 4, Kavita Rawat 5, Atul Krishna 6, Praveen K. Shukla 6, Raj K. Tripathi 5, Mohammad I. Siddiqui 4, Rabi S. Bhatta 3, Gopal Gupta 2, Vishnu L. Sharma CSIR-Central Drug Research Institute, Medicinal and Process Chemistry Division, Lucknow, India, 2 CSIR-Central Drug Research Institute, Endocrinology Division, Lucknow, India, 3 CSIR-Central Drug Research Institute, Pharmacokinetics & Metabolism Div., Lucknow, India, 4 CSIR- Central Drug Research Institute, Molecular & Structural Biology Division, Lucknow, India, 5 CSIR-Central Drug Research Institute, Toxicology Division, Lucknow, India, 6 CSIR-Central Drug Research Institute, Microbiology Div., Lucknow, India Background: The growing health and economic burden of STIs, HIV, and population growth in resource-poor countries have led to integrate them and vaginal microbicides have emerged as an option with additional advantage of women control. Methods: Fifteen N-alkyl/aryl-4-(3-substituted-3-phenylpropyl) piperazine--carbothioamide (3-27) derivatives were synthesized as topical vaginal microbicides and evaluated for anti-trichomonas, spermicidal, antifungal and reverse transcriptase (RT) inhibitory activities. To assist the sulfhydryl (SH) binding molecular mechanism of synthesized compounds, hexokinase and DTNB assays were carried out because -SH group present over sperm and Trichomonas is vital for their survival. All compounds were tested for safety through cytotoxic assay against human cervical cell line (Hela) and compatibility with vaginal flora, Lactobacillus. Docking study of most promising vaginal microbicide (3) was carried out to find out its docking position and orientation in comparison to known RT inhibitor Nevirapine. The preliminary in vivo pharmacokinetics of compound 3 was performed in female NZ-rabbits to evaluate systemic toxicity in comparison to clinically used spermicide Nonoxynol-9. Results: Compound 3 appeared as most favorable multiple active scaffold, comprising of RT inhibition (72.30%), spermicidal (MEC 0.0%), anti-trichomonas (MIC 7.78 µg/ml) and antifungal (MIC µg/ml) actions. The compound s low toxicity to HeLa cells and Lactobacillus growth would eventually favor vaginal administration. Hexokinase and DTNB assays proved the molecular mechanism involving SH binding. Compound 3 docked on RT in a position and orientation similar to the Nevirapine. In vivo pharmacokinetics of 3 suggested minimal side effects in comparison to N-9. Conclusions: The study resulted into novel compound (3) as women controlled topical vaginal microbicidal spermicide possessing RT inhibitory activity in order to empower women to deal independently with their reproductive health and fertility. IND-directed Pharmacology and Toxicology of IQP-0528, a Novel HIV- Topical Microbicide Robert W. Buckheit Jr., Christian F. Freguia, Anthony Ham, Karen W. Buckheit ImQuest BioSciences Inc., Frederick, MD, United States Background: In the absence of a vaccine, topical microbicides are considered one of the best strategies for HIV prevention. Here, we describe the development of a novel microbicide, IQP-0528, formulated into a vaginal gel for the pre-exposure prophylaxis of sexually transmitted HIV. Methods: The efficacy and toxicity of IQP-0528 was evaluated using in vitro cell-based assays. IQP-0528 vaginal gel was characterized by rheological analysis. IND-enabling safety and toxicity studies were performed as per FDA recommendations. Results: IQP-0528 possesses good chemical and metabolic stability and ease of synthesis. IQP-0528 potently inhibits the replication of all clinical HIV subtypes (except O) with an EC 50 of nm and TC 50 of approximately 400 um and is active in the presence of seminal and vaginal fluids with minimal toxicity to cell lines, explant tissues and vaginal flora. Formulation into a gel for vaginal delivery showed favorable physicochemical properties, long term stability, sub-nanomolar potency and minimal toxicity. IQP-0528 did not cause acute toxicity in mice and rats and exhibited no genotoxicity. IQP-0528 vaginal gel (up to 0% w/v concentration) dosed daily for 28 days in rats and rabbits was well tolerated with no adverse effects and minimal systemic absorption at a clinical dose (%). Vaginal tissue concentrations were in the ug range. The gel did not elicit skin sensitization reactions in guinea pigs, no teratogenic effects were observed in prenatal development studies, and no cardiovascular effects were identified. The NOAEL of the IQP-0528 vaginal gel is a dose of 0%. Conclusions: IQP-0528 is a novel and ideal clinical microbicide candidate due to its high potency, ease of manufacturing, long-term stability, low toxicity, poor systemic but favorable local drug absorption and low cost of goods. Overall our data provide a rationale for continued advancement of this molecule to human clinical trials. 378 HIV Research for Prevention 204 HIV R4P

379 Thursday, 30 October Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44.03 Preventing Drug Resistant HIV Infection in Colonic Tissue using Tenofovir and Maraviroc Combination Topical Rectal Gels Charlene Dezzutti,2, Julie Russo 2, Lin Wang 2, Brid Devlin 3, Ian McGowan,2, Lisa C. Rohan,2 University of Pittsburgh, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 3 IPM, Silver Springs, MD, United States P44.04 Preclinical Evaluation of Multi-targeting Antiretroviral Drug Based-combinations as Candidate Microbicides Carolina Herrera, Natalia Olejniczak, Javier García Pérez 2, José Alcamí 2, Charles Kelly 3, Robin Shattock Imperial College, Infectious Diseases, London, United Kingdom, 2 Instituto de Salud Carlos III, Madrid, Spain, 3 King s College London, London, United Kingdom Background: The paradigm for topical microbicide development has been a single agent product. Interest is now turning toward combination products. A % tenofovir (TFV)/0.% maraviroc (MVC) combination rectal gel has been developed. We hypothesize the combination gel should provide greater mucosal tissue protection against drug resistant virus than single agent products. Methods: Polarized colonic explants were treated apically with dilutions (:50 to :5000) of TFV only or TFV/MVC gel product along with 0 4 TCID 50 HIV- wildtype or drug resistant (DR) viruses. Wildtype JR-CSF and DR JR-CSF (K65R) and transmitter/founder viruses, wildtype THRO and DR CH077 (K65R/Y8C), were tested. After an overnight culture, the explants were washed and the basolateral medium was replenished. Every 3 to 4 days medium was harvested, stored and replenished. HIV- replication was monitored in culture supernatant by p24 ELISA. Results: For JR-CSF, TFV gel protected 5 of 6 explants at 328 µm (:00 dilution of the gel) while twice as much TFV was needed to protect explants against JR-CSF (K65R); a 2-fold difference in activity. The TFV/MVC gel was completely protective at 65.5 µm TFV/3.9 µm MVC (:500 dilution of the gel) and was more than 50% effective at preventing HIV infection at 32.8 µm TFV/2.0 µm MVC (:000 dilution) for both JR-CSF and JR-CSF (K65R). When tested against the transmitter/ founder viruses, 65.5 µm TFV/3.9 µm MVC was needed to protect all explants against THRO. However, CH077 was more resistant to the TFV/MVC protecting only 2 of 6 explants at 328 µm TFV/9.5 µm MVC (:00 dilution). Conclusions: TFV/MVC gel was 5-fold more potent than TFV gel likely reflecting the activity of MVC and it was equally potent against JR-CSF and JR-CSF (K65R). While THRO showed similar activity to the TFV/MVC gel as both JR-CSF viruses, CH077 showed marginal activity toward the TFV/MVC combination which may reflect other compensatory mutations. Additional testing is needed to confirm DR virus susceptibility to TFV/MVC gel. Background: Antiretroviral (ARV) drug combinations are highly effective in HAART and may also be more effective as microbicides than single drugs. This study aims to assess the activity of tenofovir, dapivirine and darunavir combinations against wild-type and resistant HIV-, SIVmac32H and RT-SHIV isolates Methods: Antiviral efficacy of dual and triple combinations of a nucleotide reverse transcriptase inhibitor (NRTI), tenofovir, a non- NRTIs, dapivirine and a protease inhibitor, darunavir; was evaluated. The combinations were assessed in cellular (TZM-bl cells, PM- CD4+T cells, C866 cells and activated PBMCs) and colorectal explant models. Preincubation of cells or tissue with the drugs individually or in combination, for one hour was followed by addition of virus. Wild type isolates (BaL, YU.2, SIVmac32H and RT-SHIV) and NRTI-escape mutants (point mutations K65R +/- M84V introduced in YU.2 and SIVmac32H) were used. Infection was determined by measurement of luciferase expression (in TZM-bl cells) or p24 viral antigen in culture supernatants Results: All dual combinations tested in cellular and tissular models against all isolates showed an increase of activity for at least one of the drugs included in the combination compared to the drugs used alone. The triple combination further augmented the inhibitory activity. All combination tested inhibited NRTI-resistant isolates. The introduction of point mutations affected the viral replication fitness in HIV and SIV. Interestingly, dose-response curves were affected by the preclinical model used, emphasizing the need to use multiple cellular and tissue models during screening of candidate microbicides Conclusions: The positive results obtained against HIV- and SIV/SHIV isolates allow the design of in vivo studies in macaques before clinical trials. The activity against HIV-, SIV/SHIV indicate drug combinations inhibiting reverse transcription and viral maturation have a great potential as effective microbicides able to inhibit HIV- transmission 379

380 Posters Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44.05 P44.06 Mini CD4-heparan Sulfate Mimetic Conjugates Display Sub Nanomolar Anti-HIV- Activity and Protect Macaques against a SHIV62P3 Vaginal Route Challenge Françoise Baleux, Kevin Arien 2, Delphine Desjardins 3, Jo Michiels 2, Yves-Marie Coic, Bridgette J. Connell 4, David Bonnaffé 5, Kawthar Bouchemal 6, Roger Le Grand 3, Guido Vanham 2, Nathalie Dereuddre-Bosquet 3, Hugues Lortat-Jacob 4 Institut Pasteur, unité de Chimie des Biomolécules, URA CNRS 228, Paris, France, 2 Institute of Tropical Medicine, Virology Unit, Antwerp, Belgium, 3 CEA, Service d Immunovirologie - IDMIT, Fontenay-aux- Roses, France, 4 Institut de Biologie Structurale, CNRS, CEA, Université Grenoble-Alpes, UMR 5075, Grenoble, France, 5 Université Paris-Sud, Institut de Chimie Moléculaire et des Matériaux d Orsay, Orsay, France, 6 Université Paris-Sud, Faculté de Pharmacie, Chatenay-Malabry, France Background: The HIV- gp20 coreceptor binding site (CoRBS) can be targeted by heparan sulfate (HS) or HS-like sulfopeptides, however its cryptic nature limits its vulnerability. We hypothesized that molecules comprising a CD4 mimetic covalently linked to HS-like compounds would bind gp20 through the CD4 moiety, expose the CoRBS and render it available to be blocked by HS (see Connell BJ. et al. Front. Immunol. 203). Methods: CD4-mimetics (mcd4) were conjugated to synthetic HS or to sulfopeptides (PS). A Surface Plasmon Resonance platform was developed to analyze gp20 binding to HS, CD4, and detergent solubilized CXCR4 and CCR5, functionally captured on biacore chips. Inhibition of infectivity of primary HIV- by a series of mcd4-ps was measured in a standardized assay with CD4 and CoR expressing TZMbl cells. Finally in vivo activity of mcd4-ps, formulated in hydroxyethylcellulose-based gel, was assessed with female cynomolgus macaque s model of intravaginal (IVAG) single high dose challenge with SHIV62P3. Results: We demonstrated that mcd4-conjugates efficiently block the HS, the CD4 and the CoR binding sites of gp20 and as such, mimic several of the recently described broadly neutralizing antibodies. In cell culture, mcd4-ps neutralize both R5- and X4- tropic HIV- of various clades with pico to nano molar IC 50 in the absence of cellular toxicity. Moreover, cynomolgus macaques treated with mcd4.-ps one hour before IVAG challenge with SHIV62P3 were fully protected against acquisition of infection whereas all control animals were infected. Conclusions: We engineered a new class of compounds which have the unique ability to target two critical and highly conserved regions of gp20, i.e. the CD4 and the CoR binding sites, in addition to the V2, V3 loops. Chemically defined, these 5.5 kda molecules which neutralize both R5- and X4-tropic HIV- with very low IC 50 are amenable to largescale production, and in view of their in vivo activity could be considered for the development of a microbicide approach. Engineering and Characterization of a Bispecific Entry Inhibitor for Non-vaccine Biomedical Prevention of HIV- Infection Krystal Hamorsky, Adam Husk, Nobuyuki Matoba University of Louisville School of Medicine, Owensboro Cancer Research Program, Owensboro, KY, United States Background: Entry inhibitors could offer promising means for nonvaccine biomedical prevention against HIV infection, such as topical microbicides. Here, we aim to create a novel entry inhibitor by translationally fusing a broadly HIV- neutralizing monoclonal antibody (mab) and a carbohydrate-binding antiviral protein targeting the glycan shield of viral envelope. We hypothesize that such a bispecific inhibitor may effectively block and control HIV- infection. Methods: We engineered a prototype bispecific fusion protein based on the antigen-binding fragment (Fab) of the CD4 binding site-specific mab VRC0 and the oligomannose-specific, non-cytotoxic/inflammatory/ mitogenic antiviral lectin Avaren (VRC0-Av). VRC0-Av was produced in Nicotiana benthamiana plants due to the prospective scalability and cost effectiveness of plant-based expression systems. The fusion protein was purified by a series of chromatography steps, and tested for the binding properties and anti-hiv activity by surface plasmon resonance (SPR) and Env-pseudotyped virus neutralization assays, respectively. Results: The fusion protein was efficiently purified to >95% homogeneity by a two-step chromatography process. SPR demonstrated that both active sites of the VRC0-Av fusion retained affinity to a recombinant HIV- Env protein gp20. VRC0-Av had significantly stronger HIV-- neutralizing activity than a : equimolar mixture of each component (i.e., VRC0 Fab and Avaren) and respective bivalent parental molecules (i.e., VRC0 IgG and Avaren-Fc fusion) against multiple strains. We are currently developing a dry powder formulation of VRC0-Av to enhance stability and facilitate delivery system development. Conclusions: These results highlight the advantages of VRC0-Av bispecific fusion protein over a mixture of the two original entry inhibitors in terms of efficacy, manufacturing and drug development. Our data warrant further preclinical efficacy and toxicity studies of VRC0-Av. 380 HIV Research for Prevention 204 HIV R4P

381 Thursday, 30 October Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44.07 P44.08 Counteracting Semen-mediated Enhancement of HIV Infection and Enveloped Virus Infection by a Lysine-specific Molecular Tweezer Edina Lump, Laura Castellano 2, Thomas Schrader 3, Gal Bitan 4, James Shorter 2, Jan Münch Ulm University Medical Center, Institute of Molecular Virology, Ulm, Germany, 2 Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, United States, 3 University of Duisburg- Essen, Essen, Germany, 4 University of California at Los Angeles, Los Angeles, CA, United States Background: Semen contains amyloid fibrils that markedly enhance HIV- infection by sequestering viral particles and increasing cell entry rates. Thus, counteracting this proviral activity in semen presents a novel strategy of reduce or block sexual HIV- transmission. It has previously been shown that a lysine-specific molecular tweezer (CLR0) is capable of preventing the formation of Aß and α-synuclein fibrils that are associated with Alzheimer s and Parkinson s disease. Here, we explored whether CLR0 also affects the formation and function of seminal amyloids. Methods: The effect of CLR0 on seminal amyloid formation and its degradation was studied by electron microscopy and spectrofluorometry; interaction of CLR0-treated amyloid with viral particles was analyzed by zeta potential measurements and confocal microscopy; the effect of CLR0 on amyloid- and semen-mediated enhancement of HIV, HCV, and HSV-2 infection was determined by infection assays; Results: We demonstrate that CLR0 blocks the formation of seminal amyloids and even slowly disaggregates pre-formed fibrils. Furthermore, the tweezer neutralizes the positive surface charge of the fibrils thereby preventing their interaction with virions and abrogating their HIV enhancing activity. Most importantly, CLR0 also counteracts the ability of semen to boost infection of HIV transmitted/founder viruses. Unexpectedly, CLR0 also inhibits HIV and other enveloped viruses such as HCV or HSV-2 by directly disrupting virion integrity. Conclusions: CLR0 has previously been tested safe and therapeutically active in mouse models of amyloid diseases. Thus, its low toxicity and the combined anti-amyloid and anti-viral activities make CLR0 a promising candidate for microbicide development to prevent HIV and other sexually transmitted virus infection. Intravaginal Films Delivering Aminoglycosides: Safety in a Macaque Model Dorothy L. Patton, Yvonne Sweeney, Tian Zhou 2,3, Lisa C. Rohan 2,3, Alexander M. Cole 4 University of Washington, Obstetrics & Gynecology, Seattle, WA, United States, 2 Magee Women s Research Institute, Pittsburgh, PA, United States, 3 University of Pittsburgh, School of Pharmacy, Pittsburgh, PA, United States, 4 University of Central Florida, Orlando, FL, United States Background: Aminoglycosides, which restore the natural production of retrocyclins in the cervicovaginal mucosa, enhance innate antiviral immunity. Tobramycin, an aminoglycoside antibiotic, can be delivered by dissolvable vaginal films to heighten protection against sexually transmitted HIV- infection. This study assessed the safety of intravaginally delivered tobramycin in a macaque model for topical microbicide use. Methods: In test product vs placebo studies of n=6 per group, we assessed the effects of a film formulation containing 5 mg tobramycin during two weeks of daily applications. We performed baseline colposcopy, vaginal microbiology, smear and ph measurements, followed by intravaginal film placement in each animal. Thirty minutes after film placement (test or placebo), repeat vaginal flora, smear and ph measurements were made. These procedures were repeated daily for five days in two successive weeks. On the following Monday (day 5), a follow-up exam was conducted to document recovery. Results: Colposcopic observations demonstrated that repeated exposures to the 5mg tobramycin film did not lead to serious adverse findings, i.e. cervicovaginal tissue abrasion and/or friability, in any of the 6 animals. Microbiologic assessment revealed populations of H 2 O 2 producing lactobacilli and viridans streptococci fluctuated over the course of the experiment, but no clear trend of growth or suppression was noted for either organism. The incidence of enteroccocus and E. coli remained low, yet each was more prevalent at the end of the study than at baseline. In both study arms, vaginal ph measures fluctuated modestly throughout the study, and on average decreased from neutral ph at baseline to ph at the final visit. No product related induction of neutrophil infiltration was noted by Gram stain. Conclusions: Repeated vaginal exposures to the aminoglycoside film were well tolerated in the macaque model. Studies assessing systemic absorption and distribution of tobramycin in this experiment are underway. 38

382 Posters Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44.09 P44.0 RV306, an Evaluation of a 48 Week ALVAC- HIV AIDSVAX B/E Vaccination Regimen in Thailand: Participation Rates for Optional Specimen Collections Punnee Pitisuttithum, Sorachai Nitayaphan 2, Suwat Chariyalertsak 3, Nicos Karasavvas 4, Jaranit Kaewkungwal 5, Viseth Ngauy 4, Sandhya Vasan 4, Merlin L. Robb 6, Nelson L. Michael 6, J. Kendall Brown 6, Charla Andrews 6, Benjaluck Phonrat, Jittima Dhitavat, Jean Louis Excler 6, Jerome H. Kim 6, Robert J. O Connell 4, on Behalf of the RV306 Study Team Vaccine Trial Centre, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand, 2 Royal Thai Army Clinical Research Center, AFRIMS, Bangkok, Thailand, 3 Research Institute for Health Sciences (RIHES), Chiang Mai University, Chiang Mai, Thailand, 4 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand, 5 BIOPHICS - Center of Excellence for Biomedical and Public Health Informatics, Bangkok, Thailand, 6 U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Bethesda, MD, United States Background: Vaccination with the RV44 regimen showed a waning efficacy of 60% at one year and 3.2% three years post-last vaccination. Risk correlated with plasma anti-env binding antibody responses, but specimen volumes and collection sites limited analysis. HIV-vaccine naïve volunteers were vaccinated with the RV44 regimen plus a 2 month boost for immunologic assessment in multiple systemic and mucosal compartments. Methods: Volunteers from 3 sites in Thailand were randomly enrolled into 4 placebo-controlled groups receiving the RV44 regimen plus no boost, ALVAC-HIV+AIDSVAX B/E, AIDSVAX B/E, or ALVAC at 2 months. Immune responses are being assessed both systemically and for additional site specimen collections in a subset of willing volunteers. Results: Of 63 screened participants, 362 were enrolled (48% male and 52% female) with a mean age of 28 years. Consent to provide rectal secretions was provided by 69/72 (40%) of male volunteers, ranging from 2-90% among the three sites, while 35/72 (78%) consented to provide semen. Female participants consented for cervico-vaginal secretion collection (VTC: 69/95 (73%); RTA: 39/76 (5%); RIHES: 9/9 (47%), all sites: 7/90 (62%). Volunteers in two sites also consented for a single invasive procedure per participant as follows: Cervical biopsy (VTC: 44/95 (46%); RTA: 5/76 (20%); all sites 59/7 (35%), sigmoid biopsy in males [VTC: 29/57 (5%); RTA: 3/74 (8%); total 42/3 (32%)], leukapheresis: (VTC: 30/52 (20%); RTA 2/50 (4%); total 5/302 (7%) and bone marrow aspiration [VTC: 9/52 (6%); RTA: 5/50 (3%); total: 4/302 (5%)]. Conclusions: Mucosal collections and invasive procedure participation rates were high in this clinical trial conducted in Thailand, demonstrating both favorable disposition to research participation and effective counseling at the sites. Differences in participation in these procedures among sites provide insights that may lead to strategies to improve participation in future trials. Candidate Microbicide 5-hydroxytyrosol (5- HT) Inhibits Productive R5 HIV- Infection of Human Cervical Tissue Explants (CTE) Elisa Saba, Massimo Origoni,2, Gianluca Taccagni, Claudio Doglioni,2, David Auñón 3, Eduardo Gomez-Acebo 3, Josè Alcami 4, Guido Poli,2, Elisa Vicenzi San Raffaele Scientific Institute, Milan, Italy, 2 Vita-Salute San Raffaele University, School of Medicine, Milan, Italy, 3 Seprox Biotech SL, Madrid, Spain, 4 Instituto de Salud Carlos III University of Alcala, Madrid, Spain Background: Despite the encouraging results obtained in the CAPRISA trial by using Tenofovir gel to prevent HIV- sexual transmission, the development of an effective microbicide remains a major priority. Among candidate new microbicides, we are investigating the phenolic phytochemical compound 5-HT (previously shown to be possess anti- HIV-, anti-inflammatory and anti-oxidant activities in isolated cell systems) in human cervical tissue explants (CTE). Methods: CTE established from HIV- seronegative women undergoing hysterectomy for benign tumors, were infected ex vivo with R5 HIV- BaL in the presence or absence of increasing concentrations of 5-HT or, as control, of 3TC. Productive infection of HIV- was measured in terms of p24 Gag expression in the culture supernatants. In addition, total viral DNA was quantified by RT-PCR in the different experimental conditions. Results: A decrease of p24 Gag release was observed in the culture supernatant after incubation with 200 µm of 5-HT and this effect was also associated with 50% decrease of total viral DNA, as determined on tissue blocks after 2 days of culture. No modulation of CD4 and CCR5 or of activation markers (CD69, CD25, CD38) was observed in CD4 + lymphocytes incubated with up to 200 µm of 5-HT. Ongoing experiments are aimed at evaluating the potential effect of 5-HT on the release of pro/anti-inflammatory cytokines and chemokines in CTE culture supernatants. Conclusions: These results confirm that 5-HT bears the potential to be developed as an anti-hiv microbicide as it inhibits HIV- replication in CTE, in addition to isolated cell systems, without inducing cytotoxicity or promoting T cell activation. 382 HIV Research for Prevention 204 HIV R4P

383 Thursday, 30 October Posters 44: Preclinical Evaluation of Biomedical Agents in Animal Models Tissues Explants P44. P44.2 LB Mucosal Susceptibility to HIV- and Tenofovir Protection in Explanted Cervicovaginal Tissues from Postmenopausal Compared to Premenopausal Women Andrea R. Thurman, Neelima Chandra, Nazita Yousefieh, Sharon M. Anderson, Susana Asin 2,3, Christianne Rollenhagen 2,3, Betsy C. Herold 4, Pedro Mesquita 4, Ashley Huber 4, Gustavo F. Doncel CONRAD, Eastern Virginia Medical School, Norfolk, VA, United States, 2 Department of Veteran s Affairs, Department of Microbiology and Immunology, White River Junction, VT, United States, 3 Dartmouth Medical School, Immunology and Microbiology, Hanover, NH, United States, 4 Albert Einstein College of Medicine, Infectious Diseases and Pediatrics, Bronx, NY, United States Background: In order to obtain data on the mucosal effects of tenofovir (TFV) on postmenopausal women, biologic endpoints related to HIV- acquisition in cervicovaginal (CV) explants from pre (PreM) and postmenopausal (PostM) women were compared. Methods: CV explants from PreM and PostM women were obtained within hour of elective surgery and treated for 24 hours with growth medium (GM) or 00 µg/ml of TFV. We assessed immune cell numbers and phenotype in explants. Additional tissues were infected ex vivo with HIV- BaL and p24 antigen production was assayed over 2 days. Tissue culture supernatants were assayed for inhibition of HIV- in TMZ-bl cells in vitro. Results: Based on preliminary data from ex vivo HIV- infection (mean p24 endpoints and categorical analyses), PreM and PostM tissues do not appear to show differences in their propensity to be infected by HIV. TFV protected both tissues fully. Anti-HIV activity of PostM explant supernatants demonstrated a trend (p = 0.06) toward reduced HIV inhibition (PostM=38.5 ± 36.2% v. PreM=60.8 ± 9.%). The supernatants of explants from PreM and PostM women incubated with TFV showed potent and equal activity against HIV (p = 0.6). Treated with GM, both types of explants produced similar levels of cytokines and chemokines. TFV increased the secretion of these immune mediators. PostM explants had significantly greater number of HIV- target CD4+ cells (p = 0.02) and CDa cells (p=0.03) and a trend towards higher numbers of HLA-DR+ activated cells (p = 0.06). Conclusions: PreM tissues have thicker epithelium, slightly fewer epithelial immune cells (especially CD4 and CDa), and similar basal cytokine/chemokine production. They also produce secretions that inhibited HIV infection in an in-vitro cell-based model to a greater extent. Although these conditions would theoretically endow prem mucosal tissues with higher resistance to infection, thus far, we have not seen increased susceptibility of postm explanted tissues to ex vivo HIV- infection. Tissue Permeability and Drug-drug Interactions of Darunavir in Intact Rabbit Colo-rectal Tissue Abhinav Kumar, Magda Swedrowska, Charles Kelly 2, Ben Forbes King s College London, Institute of Pharmaceutical Science, London, United Kingdom, 2 King s College London, Oral Immunology, London, United Kingdom Background: Cell culture models for studying drug disposition and drugdrug interactions are good but they cannot reflect the heterogeneous cellular coexistence typical of colo-rectal mucosa. The aim of this study was to establish an ex vivo excised tissue model which is closer to the more complex in vivo situation and apply it for studying the transport and interactions of darunavir (DRV) with other ARVs as reports suggest that combination microbicides is more effective. Methods: Colo-rectal tissue from a male New Zealand white rabbit was excised and transferred to oxygenated ice cold Kreb s buffer. A strip of the colo-rectal segment was opened with a lateral cut thus making a planar sheet, rinsed free of luminal contents with Kreb s buffer and stripped of the muscle layers before being mounted between twodiffusion half-cells containing oxygenated transport buffer solutions. The permeability of darunavir was measured; interactions with transporter inhibitor elacridar and effect of co-administration with tenofovir were evaluated in 3 independent experiments. Results: The respective absorptive and secretory permeability of DRV across rabbit colo-rectal mucosa was.8± cm/s and 4.6± cm/s (ER-efflux ratio 3). Modulation of the transport of DRV (0 µm) by elacidar ( µm) showed that absorptive transport increased by 4.4 fold and secretory transport decreased by 0.3 fold (ER 0.2). Coadministration of TFV (00 µm), with DRV resulted in.6 fold increase in absorptive direction and 0.5 fold decrease in secretory direction as compared to the transport of singlet agent (ER 0.8). Conclusions: Transport of DRV was vectorial and affected by transporter inhibitors, suggesting that DRV is a substrate of P-glycoprotein. DRV transport was affected by the presence of tenofovir. The intact colorectal tissue model is a valuable technique and this could be extended to include excised tissue samples from non-human primates as well as applied for assessing cervico-vaginal drug transport

384 Posters Posters 45: Pregnancy and Prevention of Mother to Child Transmission P45.0 Role and Trend of Maternal Antiretroviral Therapy in Preventing Mother-to-Childtransmission of HIV in Sub-Saharan Africa P45.02 Prevention of Mother-to-Child-Transmission: Antiretroviral Coverage and Access to Virological Testing during Breastfeeding Olatunji O. Adetokunboh, Mojisola Oluwasanu 2 Stellenbosch University, Division of Community Health, Cape Town, South Africa, 2 University of Ibadan, Faculty of Public Health, Ibadan, Nigeria Olatunji O. Adetokunboh, Tolulope Balogun Stellenbosch University, Division of Community Health, Cape Town, South Africa Background: Pediatric HIV infections are largely due to mother-to-child transmission (MTCT) in utero, during delivery, or via breastfeeding. The highest burden of global pediatric HIV infections are in sub-saharan Africa (SSA) where many of the countries still have high MTCT rates and new pediatric HIV infections. This study seeks to determine the role and trend of antiretroviral therapy in HIV positive women with respect to reduction in MTCT rate in the high burden countries of SSA countries since 2009 when call for virtual elimination of pediatric HIV was made. Methods: Data were obtained from the Joint United Nations Programme on HIV/AIDS (UNAIDS) 203 progress report on the Global Plan. The data were that of 2 SSA priority countries from 2009 to 202. Our analysis focused on final MTCT rates, percent of women receiving antiretroviral agents (ARVs) to prevent MTCT, percent of women or infants receiving ARVs during breastfeeding to prevent MTCT and percent of HIV positive pregnant women receiving ART for their own health. Results: The final MTCT rate reduced from 27% to 9%, p= Percentage of women receiving ARVs to prevent MTCT was 63% in 202, mean difference MD 30%, p=0.000 and the percentage of women or infants receiving ARVs during breastfeeding was 43%, MD 33%, p= The percentage of HIV positive pregnant women receiving ART was 54% in 202, MD 34%, p= The final MTCT rate was strongly negatively correlated with the percentage of women receiving ARVs to prevent MTCT (r= ; p< 0.000). Conclusions: There has been a significant increase in ART coverage among HIV infected women in SSA in the last few years. Countries that have higher ART coverage tend to have lower MTCT rates thereby preventing many new infections. However there is still lot of work to be done to achieve total elimination. Background: Sub-Saharan Africa accounts for about 90% of new HIVinfections in children with most of them contracting the infection via mother-to-child-transmission (MTCT). Transmission during breastfeeding accounts for about 40% of new infections. Access to antiretroviral (ARV) regimens by the HIV positive mothers and their children during breastfeeding will improve the chances of these children to be free of HIV infection. Ensuring early infant virological diagnosis of HIV infection will help in identifying those who are HIV-exposed but uninfected and making sure they are free of the disease during breastfeeding period. We will like to evaluate the coverage level of women or infants receiving ARVs during breastfeeding to prevent MTCT and access to virological testing in sub-saharan countries. Methods: Data were obtained from United Nations Children s Fund (UNICEF) 203 Children and AIDS Sixth Stocktaking Report. We analysed data from 2 SSA priority countries. Our focus was on the percentages of mother or child on ARVs to prevent MTCT during breastfeeding ( ) and early virological diagnosis of HIV in HIV exposed infants (202). Results: In 202, infants had early virological testing for HIV infection with South Africa recording 5.6% of the total number. The average percentage of infants tested was 32.2% with 5 of the countries recording < 50% coverage. Percentage of women or infants receiving ARVs during breastfeeding increased from 0.2% to 42.5%. The mean difference was 32.2%, 95%CI: , p = Ghana made a remarkable progress with a difference of 95% while 2 countries were at < 0% level in 202. Conclusions: The African priority countries made significant progress in the provision of ARVs for mothers and infants during breastfeeding period although some countries are still lagging behind. The level of virological testing is still very low, additional effort is needed to improve the early infant diagnosis in countries with poor coverage. 384 HIV Research for Prevention 204 HIV R4P

385 Thursday, 30 October Posters 45: Pregnancy and Prevention of Mother to Child Transmission P45.03 Ethics of HIV Prevention Research with Pregnant Women Liza Dawson NIH/NIAID, Division of AIDS, Bethesda, MD, United States P45.04 Barriers to Uptake of Polymerase Chain Reaction Testing for HIV Exposed Infants at Six Weeks among PMTCT Mothers at the State Hospital Osogbo, Nigeria Elizabeth Edoni, PMTCT Mothers Niger Delta University, Bayelsa, Nigeria Background: Pregnant women need safe and effective HIV prevention methods. In many countries, pregnant women are at high risk of HIV acquisition; countries with high rates of HIV incidence and generalized epidemics are also largely countries with high fertility rates. Also, social and structural barriers to women s autonomous decision-making may hinder pregnant women from protecting themselves. While the HIV prevention field is moving rapidly, testing prevention interventions in trials with pregnant women remains daunting. Ethical and regulatory barriers are significant, and community, provider, and individual women s perspectives are complex. The consequences of failing to conduct research with pregnant women are serious. Absent robust data, pregnant women must either eventually use these products without knowing if they are safe and effective, or avoid the products and continue to be exposed to high risks of HIV acquisition. Methods: Significant barriers and opportunities must be identified. Human research regulations place stringent restrictions on research with pregnant women. Although the heaviest burden of HIV lies in sub-saharan African countries, each with its own research regulations, US regulations have a significant impact. The US is a major funder of biomedical HIV prevention research, and any study funded by the US government must adhere to US regulations. Researchers may have difficulty meeting criteria for approval under this rule. Results: To overcome these challenges, a more robust ethical approach to research with pregnant women is needed. This paper explores the key ethical parameters that must be considered in finding an appropriate balance for inclusion of pregnant women in research, and discusses how this might fit under current regulatory standards. Conclusions: Research with pregnant must advance to extend the benefits of HIV prevention to this key population. A new and more appropriate ethical framework will help accomplish this goal. Background: Expand access to early infant diagnosis and earlier and improved pediatric treatment are essential in order to improve survival rates and health outcomes for children. Non-adherence in the first few weeks may lead to the development of the premature development of drug-resistant virus. This study therefore designed to document barriers to Uptake of Polymerase Chain Reaction Testing (PCR) and initiation of Cotrimoxazole (CTX) for HIV-exposed Infants at Six Weeks among PMTCT Mothers at the State Hospital, Osogbo, Nigeria Methods: A Case file of 30 PMTCT mothers who delivered two months prior to this study were randomly selected and reviewed. The HIV prevalence of pregnant women in this facility was 6.0%. Client information was reviewed and telephone interviews were conducted with each client. Counseling was provided to mothers to encourage a clinic visit for infant HIV PCR testing and CTX prophylaxis. Results: Few (7.8%) of the mothers were not aware that they need to bring their infant for HIV PCR testing and these women had brought their babies for immunizations but the infants were not identified as HIV-exposed. Some (36.9%) of the infants had received HIV PCR testing, but CTX was not dispensed. Reported barriers among respondents were distance to the clinic (78.9%), spouse disapproval (43.7%), and stigma and discrimination (39.0%). Conclusions: In order to improve early diagnosis and treatment of children, linking of children from PMTCT to care has to be strengthened. This calls for more recruitment and training of Staff. Delivery facilities should also have clear traceable records of the number of HIV-positive women who deliver

386 Posters Posters 45: Pregnancy and Prevention of Mother to Child Transmission P45.05 P45.06 The MTN-06 Pregnancy Registry: Baseline Characteristics of Enrollees from the VOICE Study and Reasons for Non-enrollment of Eligible Women Samuel Kabwigu, Lisa Noguchi 2, Jothi Moodley 3, Thes Palanee 4, Kenneth Kintu, Lulu Nair 5, R Panchia 6, Pearl Selepe 7, Jennifer E. Balkus 8, Kristine Torjesen 9, Jeanna Piper 0, Rachel Scheckter 9, Rohan Hazra, Richard Beigi 2 MU-JHU Research Collaboration, Kampala, Uganda, 2 MTN/Johns Hopkins University, Baltimore MD USA, Baltimore, MD, United States, 3 MRC, Durban, South Africa, 4 Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 5 CAPRISA, Durban, South Africa, 6 PHRU, Soweto, South Africa, 7 Aurum Institute, Klerksdorp, South Africa, 8 FHCRC and University of Washington, Seattle WA, WA, United States, 9 FHI 360, Durham, NC, United States, 0 US NIH/DAIDS, Bethesda, MD, United States, US NIH/NICHD, Bethesda, MD, United States, 2 University of Pittsburgh/Magee-Womens Hospital, Pittsburgh, PA, United States Background: As pregnant women are at risk for HIV and women at risk of HIV may become pregnant, it is important to assess the safety of candidate HIV prevention products in both non-pregnant and pregnant women. Methods: The MTN-06 pregnancy registry is a prospective observational cohort in which participants who became pregnant during MTN effectiveness studies or those with planned exposures in pregnancy safety studies are monitored for adverse obstetric outcomes; infants from these pregnancies are followed through the first year of life. Registry enrollment systematically excludes termination of pregnancy and early pregnancy loss, including early non-viable pregnancies with a transient positive test at a monthly visit, as these data are captured in parent protocols. For VOICE participants enrolled into the registry from Uganda, Zimbabwe and South Africa, we describe baseline demographic characteristics and key reasons for non-enrollment as reported by site investigators. Results: Among 5029 VOICE participants with over 5425 person-years (py) of follow-up, there were 424 pregnancies (7.8/00 py) and 20 live births. Of these pregnancies, 26 (62%) were eligible for MTN-06. The average age of women who became pregnant during VOICE was 24, with 24% of women married at baseline. Of these, 23/26 (82%) women and 85/20 (92%) of their infants enrolled in MTN-06 (average maternal age 25 years, 33% married). Reasons for non-enrollment of eligible participants into MTN-06 included additional study visit burden and employment. Cultural customs related to temporary relocation to rural areas during the postnatal period and the limitation of public access to newborns were also cited as barriers to enrollment. Conclusions: In this pregnancy exposure registry for candidate HIV prevention agents, the majority of eligible women from VOICE and their infants were enrolled. Study visit burden and local cultural customs may impact the enrollment of mothers and their infants into a pregnancy registry protocol. Impact of ARVs and Public Health Awareness on the Trend of HIV Infections among Infants Born to HIV Infected Mothers Olipher Makwaga, Anne Muigai 2, Freda Andayi, Tom Mokaya, Matilu Mwau 3 Kenya Medical Research Institute - CIPDCR, Busia, Kenya, 2 Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya, 3 Kenya Medical Research Institute - CIPDCR/Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya Background: Although ARVs and public awareness is being used to eliminate new HIV infections among infants, limited information indicating their impact on the trend of HIV prevalence among infants born to HIV infected mothers exists. This study determined the trend of HIV infection among infants born to HIV infected mothers in relation to specific ARV administration. Also assessed the knowledge about HIV transmission between mothers and infants. Methods: In a cross-sectional study, dried blood spot samples (200,n=420; 20,n=4093; 202, n=4686; 203, n=3080) collected from infants aged 8 months. These samples were analyzed at KEMRI- CIPDCR using PCR assay. Some mothers who received public health education were pregnant. All mothers were put on anti-retroviral drugs during and after delivery. Results: Out of the total number of samples tested 9.7%; 8.5%; 7.9%; 7.2% were HIV positive in 200; 20; 202; 203 respectively. The prevalence of HIV in infants whose mothers had been put on ARVS was as follows: AZT+NVP+3TC was 0.4%, 9.%, 6.3%, 6.0% in 200; 20; 202, 203 respectively. HAART was 7.%; 6.%; 4.7%; 3.9% in 200; 20; 202, 203 respectively. SdNVP was 0.3%; 7.4%; 6.2%; 6.0% in 200; 20; 202; 203 respectively. Those mothers who had not been given any ARV had their infants with HIV prevalence reaching.2%; 2.6%; 2.7%; 2.9% in 200; 20; 202; 203 respectively. Those mothers who were aware that ARVs reduce transmission of HIV from mother to child were 30%vs70%; not breastfeeding were 37%vs8% and Delivery at the hospital were 43%vs89% before and after public health awareness respectively. Conclusions: Direct relationship between specific ARV administration and HIV infection among infants exists. This is evident by the fact that HIV prevalence appeared to decrease with subsequent years of provision of specific drugs. HAART seemed to provide greater impact in HIV prevention compared to others. There is need for intensified awareness among reproductive women to help in reduction of transmission. 386 HIV Research for Prevention 204 HIV R4P

387 Thursday, 30 October Posters 45: Pregnancy and Prevention of Mother to Child Transmission P45.07 P45.08 Obstetric and Infant Outcomes Following Planned Maternal Third Trimester Exposure to Tenofovir % Vaginal Gel Lisa M. Noguchi,2, Joseph Biggio 3, Katherine Bunge 2,4, James Dai 5, Karen Isaacs 6, Kristine Torjesen 6, Samuel Kabwigu 7, Jill Schwartz 8, Juan Vargas 9, Fernando Scaglia 0, Cindy Jacobson 2, D H. Watts, Jeanna M. Piper 2, Richard H. Beigi 2,4, for the MTN- 002, MTN-008 & MTN-06 (EMBRACE) Study Teams Johns Hopkins Bloomberg School of Public Health, Epidemiology, Baltimore, MD, United States, 2 Microbicide Trials Network, Pittsburgh, PA, United States, 3 University of Alabama at Birmingham, Birmingham, AL, United States, 4 University of Pittsburgh/Magee-Womens Hospital, Pittsburgh, PA, United States, 5 Statistical Center for HIV/AIDS Research and Prevention, Seattle, WA, United States, 6 FHI 360, Durham, NC, United States, 7 MU-JHU Research Collaboration, Kampala, Uganda, 8 CONRAD/EVMS, Arlington, VA, United States, 9 San Francisco General Hospital/UCSF School of Medicine, San Francisco, CA, United States, 0 Baylor College of Medicine, Houston, TX, United States, Office of the Global AIDS Coordinator, U.S. Dept. of State, Washington, DC, United States, 2 US NIAID/Division of AIDS, Bethesda, MD, United States Background: Thorough drug safety evaluation includes assessing potential impact of use on obstetric (OB) and infant outcomes. The MTN- 06 study is the first pregnancy exposure registry for anti-hiv PrEP and microbicide agents. We evaluated OB and infant outcomes for registrants enrolled from third trimester TFV gel exposure studies. Methods: Data were restricted to registrants enrolled from studies with planned TFV vaginal gel exposure: MTN-002 (open label, single dose prior to cesarean) and MTN-008 (2: placebo-controlled, 7-day use). Registry study visits occurred before delivery when possible, and at <,, 6 and 2 months for infants. Infant malformation endpoints were determined by geneticists via independent review of physical exam (PE) and photo data. Results: All 6 MTN-002 and 90% (88/98) of MTN-008 mothers were registered, with 25% (n=4) of MTN-002 and 97% (n=86) of MTN-008 participants enrolling prior to known pregnancy outcome. Demographics were similar for MTN-008 enrollees and non-enrollees in the registry. Infant retention at 2 months was 88% (MTN-002) and 80% (MTN-008). One defect (ear canal) was noted in MTN-002, a rate (6%) comparable to the 3% US background rate for malformations (p=0.5); no defects were noted in infants from MTN-008. Compared to placebo (n=30), TFV gel (n=58) was not associated with preterm delivery (/58 (2%) vs. 2/30 (7%), p=0.27), postpartum hemorrhage (/58 (9%) vs. 3/30 (0%), p=0.36), non-reassuring fetal status (3/58 (5%) vs. /30 (3%), p=.0), chorioamnionitis (/58 (2%) vs. 2/30 (7%), p=0.27), gestational diabetes (0/58 (0%) vs. /30 (3%), p=0.34), or abnormal infant PE findings in the first year of life (4/58 (24%) vs. 8 (27%), p=.0). Conclusions: This first report from a novel pregnancy registry suggests third trimester TFV gel exposure is not associated with infant malformation or adverse OB or infant outcomes. Future HIV chemoprevention studies should include safety evaluation, including registry participation, for pregnant mothers and their infants. A Case Control Study of Factors Associated with HIV Infection Despite Overall Low Transmission Rates in HIV Exposed Infants in Rural Kenya Nicollate Awuor Okoko, Kevin Owuor, Jayne Lewis Kulzer,2, George Owino, Irene Ogolla, Ronald Wandera 3, Elizabeth Anne Bukusi,2, Craig R. Cohen,2, Lisa Abuogi,4 Family AIDS Care and Education Services (FACES), Research Care and Training Program (RCTP), Centre for Microbiology Research (CMR), Kenya Medical Research Institute (KEMRI), Kisumu, Kenya, 2 University of California San Francisco (UCSF), Departments of Obstetrics, Gynaecology and Reproductive Sciences; Medicine, San Francisco, CA, United States, 3 Rongo District Hospital, Ministry of Health, Rongo, Kenya, 4 University of Colorado, Department of Paediatrics, Denver, Denver, CO, United States Background: Despite the availability of Prevention of Mother-to-Child HIV Transmission (PMTCT) interventions and donor and government investments in developing country implementation, vertical HIV transmission persists. In Kenya, an estimated 37,000 to 42,000 infants are infected with HIV annually due to mother-to-child transmission (MTCT). This study explored the reasons for MTCT persistence in areas with overall low transmission rates and PMTCT service provision. Methods: A case-control study of HIV-exposed infants (HEI) enrolled in follow-up care between January and June 202 was conducted at 20 rural health facilities supported by Family AIDS Care and Education Services (FACES) in Nyanza Province, Kenya. Cases were HEI who turned HIV positive, controls were HEI who remained negative at last test; an equal number of controls were randomly selected after matching based on birth month and gender. Data were abstracted from routine PMTCT registers, HEI cards, and infant forms. Logistic regression analysis was conducted to determine factors associated with HIV infection. Results: Forty-five cases and 45 controls were compared. Maternal, clinical and infant factors associated with HIV-infected infants included poor PMTCT service uptake including late enrolment of infant to follow up, (OR = 0.4, 95%CI: ), poor infant prophylaxis adherence (OR=8.32, 95%CI ), and fewer antenatal (ANC) visits (OR = 0.62, 95% CI: ). Mothers of cases were significantly less likely to report having received clinic level HIV education and counseling compared to controls (OR 0.3, 95%CI and OR 0.2, 95% CI ). Maternal disclosure, gestation at first ANC visit, and infant feeding type were not significantly associated. Conclusions: Poor PMTCT service uptake and a reported absence of clinic level HIV education and counseling were associated with MTCT. More emphasis on PMTCT service provision including counseling and education are needed to minimize HIV transmission to infants

388 Posters Posters 46: PrEP Trials: Preparing for Demos, Participant Experiences P46.0 Preparing for Risk-reduction Counseling on Pre-exposure Prophylaxis for Women in Kenya and South Africa Amy Corneli, Emily Namey, Khatija Ahmed 2, Kawango Agot 3, Jennifer Headley, Kevin Mckenna, Joseph Skhosana 2, Jacob Odhiambo 3 FHI 360, Durham, NC, United States, 2 Setshaba Research Centre, Soshanguve, South Africa, 3 Impact Research and Development Organization, Kisumu, Kenya P46.02 Guidelines on Informed-Choice Counseling for Women Using Pre-exposure Prophylaxis (PrEP) Amy Corneli, Irina Yacobson, Emily Namey, Kawango Agot 2, Khatija Ahmed 3, Joseph Skhosana 3, Jacob Odhiambo 2 FHI 360, Durham, NC, United States, 2 Impact Research and Development Organization, Kisumu, Kenya, 3 Setshaba Research Centre, Soshanguve, South Africa Background: Our previous survey research on pre-exposure prophylaxis (PrEP) and risk compensation with women at HIV risk in Bondo, Kenya, and Soshanguve, South Africa, showed that a considerable minority of women may engage in riskier sexual behavior if taking PrEP. We conducted a small qualitative study to explore perceptions of an informed-choice approach to PrEP counseling. This approach promotes condoms but empowers women who are unable or unwilling to use condoms to make informed decisions regarding their sexual health when taking PrEP. Methods: We conducted eight focus groups with women in Bondo and Soshanguve (four per site). We introduced the counseling approach and presented participants with multiple messages on PrEP and on risk compensation when using PrEP, with some concepts framed as gain versus loss. Responses were coded on aspects of the messages that were perceived to be effective and ineffective and on perceptions of the counseling. Themes were identified and summarized. Results: Preliminary analysis suggests that participants appreciated the counseling s autonomy framework and believed messages on risk compensation would be understood by women in their communities. Participants generally liked the positive tone of the gain-frame messages and found them polite and encouraging. The loss-frame messages were viewed as harsh and threatening, but powerful. Many believed these messages cautionary tone and description of consequences would more effectively influence women s behavior, primarily in Soshanguve. In Bondo, many participants preferred the softer gain-frame messages and described clarity and comprehension as critical aspects of messaging. Conclusions: Both gain- and loss-frame messages were generally understood, although variation existed in their acceptability and perceived influence. Research among actual PrEP users is needed to understand the effectiveness of message framing and of the informedchoice counseling approach, particularly in African contexts. Background: Our previous survey research with women at risk of HIV infection in Kenya and South Africa showed that a considerable minority of women may engage in riskier sexual behavior if they take PrEP. Enhanced prevention efforts are needed to minimize risk compensation and promote informed decision-making on sexual health when using PrEP. Methods: We systematically reviewed best practices in informed choice counseling and considered how they could be applied to strengthen the decision-making process within HIV risk-reduction counseling. We also integrated essential knowledge and key counseling messages to support sexual health decisions about PrEP based on findings from a study on PrEP and risk compensation. Results: Counseling guidelines were developed for African women who are considering initiating, continuing, or stopping PrEP. The guidelines are intended to be used in conjunction with evidence-based behavioral change strategies and are framed using a decision tree approach. They take a woman s individual needs and circumstances into consideration and guide her through the decision-making process by asking essential questions and communicating key sexual health messages. The guidance encourages condom use while on PrEP, but also empowers women who are unable or unwilling to use condoms consistently to make betterinformed decisions by considering contraception, considering regular screening and treatment for STIs, and placing additional emphasis on PrEP adherence. Guidance for maintaining sexual health is also provided for women who transition off of PrEP. Conclusions: Risk-reduction counseling for women considering, continuing, or stopping PrEP must acknowledge that consistent condom use may not always be a realistic option. These women need specific knowledge and individualized messages to inform their sexual health decisions. The counseling guidelines will be disseminated to PrEP demonstration projects and country governments considering the approval of PrEP. 388 HIV Research for Prevention 204 HIV R4P

389 Thursday, 30 October Posters 46: PrEP Trials: Preparing for Demos, Participant Experiences P46.03 Perceptions and Practice of Heterosexual Anal Sex amongst VOICE Participants in Uganda, Zimbabwe and South Africa Zoe Duby,2, Miriam Hartmann 3, Elizabeth T. Montgomery 3, Christopher J. Colvin, Barbara Mensch 4, Ariane van der Straten 3 University of Cape Town, School of Public Health, Cape Town, South Africa, 2 Desmond Tutu HIV Foundation, Cape Town, South Africa, 3 RTI International, San Francisco, CA, United States, 4 Population Council, New York, NY, United States P46.04 Preparing for PrEP & Immediate Treatment: Focus Group Discussions in Advance of a Demonstration Project in South Africa Robyn Eakle,2, Goitse Manthata, Jonathan Stadler, Judie Mbogua, Maria Sibanyoni, W.D. Francois Venter, Helen Rees Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 2 London School of Hygiene & Tropical Medicine, Infectious Disease Epidemiology, London, United Kingdom Background: In VOICE, a phase IIB study testing vaginal gel and oral tablets for HIV prevention, 5029 female participants in 3 sub-saharan African countries used Audio Computer-Assisted Self-Interviewing (ACASI) to report adherence to study products and sexual behaviour. At baseline 7% of participants reported anal intercourse (AI) in the past 3 months. MTN-003D, a qualitative ancillary study, explored reasons, motivations and context of engaging in AI and rectal gel use. Methods: In-depth interviews (IDIs) were conducted in participants preferred language (Luganda, Shona, Zulu or English) with 88 systematically selected exited VOICE participants (26 Zimbabwean, 22 Ugandan and 40 South African). Results: Perceptions of heterosexual AI practice varied between countries. While AI was generally perceived to be male initiated, perceptions of women s ability to refuse AI, and narratives of pleasure and pain during AI differed between countries. Participants listed motivating factors for AI including male pleasure, relationship security, self-perceptions of having an over-stretched or over-lubricated vagina, coercion and financial incentives. Findings suggest low levels of condom use for AI and poor knowledge of condom compatible lubricant. One participant disclosed using study gel rectally; and one participant cited the belief that vaginal use of gel would provide protection for AI. Reactions to AI questions elicited varied responses amongst participants, ranging from shock, disgust, embarrassment to amusement. Conclusions: Findings provide insight into the under-researched taboo topic of heterosexual AI in sub-saharan Africa, shedding light on relationship contexts and gendered power dynamics in which AI takes place. Describing attitudes towards and practices surrounding heterosexual AI such as condom and lubricant use, the findings inform current HIV prevention priorities aimed at women in these countries, as well as future prevention efforts to address HIV transmission through AI, such as rectal microbicides. Background: The WHO and others have called for demonstration projects to understand the deliverability of ARV-based prevention methods. One project will be in South Africa through The Sex Worker Project, offering oral PrEP and immediate treatment to female sex workers (FSWs). In advance of the demonstration project, information will be collected to gain insight into how these interventions can be effectively delivered to FSWs. Methods: Focus Group Discussions (FGDs) are currently being conducted amongst FSWs at 2 demonstration project clinics. Participants are being recruited from the clinic and surrounding communities. FGDs are exploring FSWs perceptions of: optimal prevention and treatment delivery, attitudes towards & perceptions of the use of oral PrEP and immediate treatment, how a comprehensive HIV prevention & treatment programme could function as an incentive to access services, & the acceptability & feasibility of regular HIV testing every three months. Results: Preliminary data from FGDs and local stakeholder consultations point to several themes to be considered for the demonstration project, and future implementation. Risk disinhibition is a concern, as some FSWs could stop using condoms. This may affect the market rates of sex for money, as those not using condoms could charge higher prices. HIVpositive FSWs feared that clients could favour those on PrEP in instances where no early treatment was available. Possible resistance, in the case of inconsistent adherence, was also a concern. Women felt positive about the proposed service delivery mechanism, including the frequency of testing and were willing to participate even without financial incentives. Conclusions: The FGDs raised important issues for consideration. It will be crucial to develop careful messaging, especially on the importance of adherence and consistent condom use as combined protection. Overall, FSWs have been positive about the deliverability of PrEP and immediate treatment, with the right messaging

390 Posters Posters 46: PrEP Trials: Preparing for Demos, Participant Experiences P46.05 Characteristics of HIV Discordant Couples who Separated while Enrolled in a HIV Prevention Clinical Trial in Kenya P46.06 Following the Doctor s Advice: Experiences of HIV Serodiscordant Couples Enrolled in a PrEP Demonstration Project in Kenya Lawrence Mwaniki Mwihaki, Elizabeth Mugoiri Irungu 2, John Njoroge Mwathi 3, Snaidah Ongachi Ayub 3, Kenneth Kairu Ngure 4, Nelly Rwamba Mugo 4 Parters in Prevention Thika- University of Washington, Data, Nairobi, Kenya, 2 Parters in Prevention Thika- University of Washington, Clinic, Nairobi, Kenya, 3 Parters in Prevention Thika- University of Washington, Counseling, Nairobi, Kenya, 4 Parters in Prevention Thika- University of Washington, Managerial, Nairobi, Kenya Background: Relationship dissolution among HIV serodiscordant couples enrolled in clinical trials bears a negative impact on adherence to study product and retention to clinic visits. We evaluated characteristics of couples that separated while enrolled in HIV prevention clinical trial in Kenya Methods: We conducted univariate analysis to determine factors among HIV discordant couples associated with ever separating while enrolled at the Thika site in the Partners PrEP Study, a randomized controlled trial conducted in Kenya and Uganda. Participants reported relationship status at each visit. Data on relationship stability was extracted from the counselors chart notes. Results: Of the 496 HIV discordant couples enrolled, 98 (20%) reported ever separating. In relationships reporting separation, HIV uninfected persons were 34 [IQR 28, 39] years, 77% were male and duration of relationship was 3 [IQR, 7] years. Compared to couples with children together, those without were likely to separate (30.% vs 5.0%, OR=2.4, 95%CI.6-3.8, p< 0.000). Couples who did not consider themselves to be married (36.4% vs 8.6%, OR=2.5, 95%CI.2-5.2, p=0.06) and couples who had known their discordant status for less than a year at baseline (2.7% vs 3.%, OR=.8, 95%CI.0-3.4, p=0.05), were more likely to separate. Compared to monogamous relationships, people in polygamous unions were twice as likely to separate (OR=.9, 95%CI , p=0.00). Couples who had stayed together longer were less likely to separate (P=0.00). Conclusions: A significant proportion of HIV discordant couples enrolled in HIV prevention trials are likely to separate. The impact of this separation on retention to care should be explored. Kenneth Ngure,2, Kathyrn Curran 3, Sophie Vusha 4, Maria Ngutu 4, Nelly Mugo 2,5, Connie Celum 2,6,7, Bettina Shell-Duncan 8, Renee Heffron 2, Jared M. Baeten 2,6,7 Jomo Kenyatta University of Agriculture and Technology, Institute of Tropical Medicine and Infectious Diseases, Nairobi, Kenya, 2 University of Washington, Global Health, Seattle, WA, United States, 3 World Health Organization (WHO), HIV/AIDS, Geneva, Switzerland, 4 Partners in Health Research and Development, Thika, Kenya, 5 Kenya Medical Research Institute, Center for Clinical Research, Nairobi, Kenya, 6 University of Washington, Epidemiology, Seattle, WA, United States, 7 University of Washington, Medicine, Seattle, WA, United States, 8 University of Washington, Anthropology, Seattle, WA, United States Background: Pre-exposure prophylaxis (PrEP) for HIV-uninfected persons is highly efficacious for HIV prevention. Evidence-based social science research is urgently needed to inform PrEP rollout. We used qualitative methods to gather insights into couples early experiences of using PrEP within a PrEP demonstration study. Methods: From June-December 203, we conducted 6 in-depth dyadic interviews with heterosexual HIV discordant couples participating at the Thika, Kenya site of the Partners Demonstration Project, a project evaluating uptake of and adherence to PrEP and ART. PrEP is offered when couples enroll in the study until the HIV-infected partner initiates ART and achieves viral suppression [i.e. as a bridge ]. We developed and applied deductive and inductive codes and identified key themes related to early experiences of initiation and use of time-limited PrEP. Results: Of the couples interviewed, all HIV-uninfected partners had initiated PrEP and 7 of the 6 HIV-infected partners interviewed had initiated ART at time of interview. Prior to joining the study, no couples had heard of PrEP. Interviewees often described their decision to take PrEP as following the doctor s advice, and believed that doctor knows best due to the trust that had been established between the health providers and couples. The friendly environment at the clinic reportedly enabled a majority of the couples to make shared decision to initiate PrEP. Many couples reported that PrEP could reduce the risk of HIV transmission, meet their aspirations for fertility and cope with HIV discordance (i.e. a solution to their discordance challenges). Remaining HIV negative in the follow-up visits reinforced couples decisions and motivations to continue adhering to PrEP. Conclusions: Among these early adopters of PrEP for HIV prevention, confidence in provider s advice and patient-friendly services were critical to decisions to initiate PrEP. PrEP responded to couples values and preferences for reducing their HIV risk. 390 HIV Research for Prevention 204 HIV R4P

391 Posters 47: Resource Tracking and Economics P47.0 Thursday, 30 October P47.02 Tracking Investments and Expenditures in HIV Prevention Research and Development: Sustaining Funding in a Shifting International Development Landscape Emily D. Donaldson, Kevin Fisher, Reuben Granich 2, Thomas Harmon 3, Polly Harrison, Arne Post Uiterweer 3, Mitchell Warren, HIV Vaccines & Microbicides Resource Tracking Working Group AVAC, New York, NY, United States, 2 UNAIDS, the Joint United Nations Programme on HIV/AIDS, Geneva, Switzerland, 3 International AIDS Vaccine Initiative, New York, NY, United States Background: Since 2004, the HIV Vaccines and Microbicides Resource Tracking Working Group has employed a comprehensive methodology to track trends in research and development (R&D) investments and expenditures for biomedical HIV prevention, including HIV vaccines, microbicides, PrEP, treatment as prevention and medical male circumcision. Methods: R&D data were collected on annual disbursements by public, private and philanthropic funders for product development, clinical trials and trial preparation, community education and policy advocacy efforts to estimate annual investment in HIV prevention R&D. Investment trends were assessed and compared by year, prevention type, research phase, funder category and geographic location. Results: In 203, investment in HIV prevention research reflected decreased public sector budgets, revised investment strategies by philanthropic donors and a continued retreat from prioritizing HIV prevention by industry. The Working Group collated and analyzed 203 data for all areas of HIV prevention R&D. International development agency funding trends were analyzed and disaggregated by research type showing a move towards funding large-scale clinical trials and implementation research, following the research pipeline and trends towards results-based financing. Conclusions: Shifts taking place in 203 in the international development landscape in both policy and strategy may have profound effects on HIV prevention research funding. The reorganization of Canada s and Australia s development agencies into their respective departments of foreign affairs and trade and the trend towards country-ownership models for the HIV/AIDS response could have profound effects on these department s priorities. It is increasingly important to ensure continued prioritization of HIV prevention research on the global development agenda by understanding and evaluating research in the context of public, private and philanthropic funding. Local Investment in HIV Prevention Research: The Contributions of African Countries that Host Biomedical HIV Prevention Clinical Trials Emily D. Donaldson, Kevin Fisher, Mitchell Warren, Julien Burns AVAC, New York, NY, United States Background: The HIV Vaccines & Microbicides Resource Tracking Working Group collects annual data on trends in research and development (R&D) investments and expenditures for biomedical HIV prevention options, including HIV vaccines, microbicides, PrEP, treatment as prevention and medical male circumcision from public, private and philanthropic sources predominantly in donor countries. There is a need to track all investments in research, including those from countries and communities in which trials take place. Methods: Investment and expenditure data were collected from countries in sub-saharan Africa which undertake HIV prevention R&D. Data were collected on annual disbursements by public, private and philanthropic funders for product development, clinical trials and trial preparation, community education and policy advocacy. Data were also collected on non-monetary investments in-kind and trial volunteer commitment to report on the full amount invested in HIV prevention R&D. Policy and environmental considerations were analyzed by country. Results: With numerous clinical trials and demonstration projects taking place in countries across sub-saharan Africa and nearly 70 percent of all HIV prevention trial participants located in these countries, the region has invested considerably in HIV prevention R&D. Promising results in pre-exposure prophylaxis, treatment as prevention and adult male circumcision have pushed these countries to support studies that will aid in the rollout and uptake of new prevention modalities. Conclusions: HIV prevention R&D could not advance without the contributions of the countries in which trials take place, including financial, collaborative and especially, the commitment of trial participants and communities. As the state of donor funding is uncertain, engagement and investment of funders and end-users from the global South in the outcomes of R&D taking place in their countries is imperative to advance the science needed to discover, develop and deliver new HIV prevention options. 39

392 Posters Posters 47: Resource Tracking and Economics P47.03 Resource Tracking to Ensure Accountability and Sustainability: HIV Prevention, Treatment, Diagnostics and Cure Research Investment Analysis Emily D. Donaldson, Kevin Fisher, Mitchell Warren AVAC, New York, NY, United States P47.04 Cost Benefit Analysis of the Centralisation of Pharmacy Services in a Multi-study Clinical Research Site in Hillbrow, Johannesburg Clare Dott, Krishnaveni Reddy, Thesla Palanee, Helen Rees Wits Reproductive Health and HIV Institute, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa Background: Since 2004, AVAC has tracked resource flows in biomedical HIV prevention research and development (R&D) as the secretariat of the HIV Vaccines & Microbicides Resource Tracking Working Group. As the field evolves, AVAC employs a comprehensive methodology to track investments and expenditures across the HIV prevention field, from basic science to implementation research. By forming partnerships with HIV advocacy, research and funding organizations to ensure accuracy in reporting and grant allocation, AVAC expanded its resource tracking efforts to including investments towards an HIV cure, treatment as prevention, therapeutics and diagnostics. Methods: Annual investment data are collected from public, private and philanthropic funders, as well as principle investigators and scientific experts, for product development, clinical trials and trial preparation, implementation research, community education and policy advocacy efforts. Investment trends are compared by year, prevention technology type, research phase, funder category and geographic location. Partnerships with public and private agencies and organizations enable close review and analysis. Results: By employing a continuous and consistent methodology, forming partnerships and expanding its data collection areas AVAC has overcome the challenges of collection of funding information and maintained over a decade of investment data. Methods stayed consistent since 2004 allowing for accurate and consistent year-to-year comparisons of grants and overall investment figures. Conclusions: Resource tracking data is an important tool for advocates and policymakers to ensure accountability with public, philanthropic and industry funders. As donor funding flatlines and industry involvement in HIV research declines, it is increasingly important to ensure continued prioritization of HIV science on the global development and research agenda by understanding and evaluating research in the context of public, private and philanthropic funding. Background: In financially constrained research settings, it is critical to explore and implement methods for optimising resource allocation. The Wits RHI Clinical Research Site was renovated with the explicit intention of accommodating multiple concurrent studies under one roof, reducing duplicative effort and maximising resource use while providing quality care to the population it serves. Key to the design efficiency has been centralisation of pharmacy services across several studies. While clinical services are often study-specific, pharmacy services provide broad coverage and can be shared. Methods: The Wits RHI Research Centre Pharmacy services several ongoing studies including MTN 020, MTN 05, MTN 06, HARP and HIM- SA. Operational expenses are fixed regardless of the number of studies serviced by the pharmacy. While only MTN 020 is an Investigational New Drug (IND) study, participants enrolled in all five studies are provided with non-ind medications by the pharmacy. The fixed operational expenses of this multi-study pharmacy were investigated for 203 by reviewing expenses incurred for staffing, pharmacy licence, electricity, maintenance, calibrations and temperature monitoring. Results: The fixed operational costs for the pharmacy were ±R (±$80 000). If an additional pharmacy was established, operational costs would be approximately doubled, as most of these costs would be duplicated. By sharing pharmacy services, the consequent economies of scale are significant, where cost per unit of work output decreases with an increase in scale as fixed costs are spread out over several studies and operational efficiency is increased. Conclusions: Centralisation of Pharmacy Services significantly reduces costs when the service is shared across multiple budgets. It also allows for the rolling over of studies, where new studies can start before older studies have ended and staff can be retained. In addition, it reduces study staffing requirements, as pharmacy staff can be cross-trained on all studies. 392 HIV Research for Prevention 204 HIV R4P

393 Posters 47: Resource Tracking and Economics P47.05 Thursday, 30 October P47.06 Getting Multi-purpose Prevention to Women Now! A Business Case for Female Condoms Is a HIV Vaccine a Viable Option in South Africa and at What Cost? Anna Forbes,2, Sarah Thurston Global Health Visions, Brooklyn, NY, United States, 2 Independent Consultant, Kensington, MD, United States Background: In 202, the UN Commission on Life-saving Commodities for Women and Children cited female condoms (FCs) as a high-impact tool that, more widely used, could save millions of lives. Pilot projects show that creating sustained demand for FCs is feasible. With strategic investment, they could become as familiar as anti-malarials, oral rehydration salts, male condoms, and other products widely used in the developing world. This business case, commissioned by the Universal Access to Female Condoms Joint Programme, shows the economic impact that such investment could have in reducing health care costs and lost economic productivity associated with unprotected sex. Methods: The business case uses peer-reviewed, open-source modelling tools to examine the return on investment (ROI) achievable by funding FC programmes and the cost effectiveness of this investment. With key data from four countries, it assesses the impact that providing one million FCs (and the requisite marketing) could have on population health status and real economy variables. Desk research and interviews also shed light on the likely economic implications across society of such investment. Results: Excellent ROI was shown for Cameroon, Nigeria and Kenya. In Myanmar, the ROI fell short of total costs due to lower HIV prevalence. Using WHO-CHOICE recommended cost effectiveness thresholds, FCs offered a highly competitive cost per DALY in all four countries, compared to other investments. FCs provide wide-reaching economic and social benefits by improving method choice and coverage, supporting women s empowerment and reducing health disparities. Conclusions: Broad macroeconomic benefits are associated with higher rates of protected sex, lower fertility, and reduced HIV prevalence, all achievable through greater access to a broad basket of contraceptive choice, including FCs. Investment in female condoms now can unleash a virtuous cycle of increased utilization, sustained demand, better health outcomes, and stronger economies. Nishila Moodley,2, Glenda Gray, Alex Welte 3, Melanie Bertram 4 Perinatal HIV Research Unit, Vaccine Research Centre, Johannesburg, South Africa, 2 School of Public Health, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa, 3 The South African Department of Science and Technology/ National Research Foundation (DST/NRF) Centre of Excellence in Epidemiological Modelling and Analysis (SACEMA), University of Stellenbosch, Stellenbosch, South Africa, 4 Health Systems Governance and Finance, World Health Organization, Geneva, Switzerland Background: South Africa is the epicentre of the global HIV epidemic. Improved access to anti-retroviral therapy has done little to counter balance the 2.5 million people infected with HIV annually. Competing disease burdens, such as trauma, place considerable demands on limited South African health budgets. While vaccination may be the most effective way to prevent infectious disease, it is not without challenges or an opportunity cost. The study aims to define the maximum HIV vaccine price that would prove cost-effective in South African public health sector. The sensitivity of cost-effectiveness to vaccine characteristics and risk-behaviour changes among vaccine recipients will be assessed. Methods: Markov models will estimate the costs and health outcomes of existing South African HIV treatment and prevention strategies with and without HIV vaccination. Estimates will be based on a lifetime horizon period and employ a governmental perspective. Cost utility analysis will be conducted with uncertainty analysis of assumed parameters. Uncertainty will be assessed by one-way sensitivity and probabilistic sensitivity analysis. Expected value of perfect information (EVPI) analysis will determine the importance on individual parameters. Results: The analysis will indicate the price at which the HIV vaccine will become most cost-effective in the South African setting and will explore the factors that contribute the most to these costs. Conclusions: The methodology will allow for the evaluation of health related interventions not available in healthcare settings. Apart from HIV vaccine research value, the study will advise decision-makers on the economic impact of HIV vaccine adoption into policy

394 Posters Posters 47: Resource Tracking and Economics P47.07 Exploring the Impact of and Requirements for Adding a Vaccine to the Updated UNAIDS Investment Framework to End AIDS Arne Näveke, Emily Donaldson 2, Chaitra Gopalappa 3, Kevin Fisher 2, Thomas Harmon, Margaret McGlynn, John Stover 3, Mitchell Warren 2 International AIDS Vaccine Initiative, New York, NY, United States, 2 AVAC, New York, NY, United States, 3 Futures Institute, Glastonbury, CT, United States Background: An updated 20 UNAIDS Investment Framework reflecting 203 WHO treatment guidelines modeled how scaling up available treatment and prevention interventions, and adding new prevention tools, including a vaccine, could reduce new HIV infections and AIDS-related deaths until This study explores the specific impact of an AIDS vaccine depending on product and program characteristics. It also explores cost-effectiveness as a major criterion for policy makers to consider a new tool for public health programs. Methods: We used the Goals model to project the impact of vaccines of various characteristics in 24 low- and middle-income countries (LMICs) for various scenarios of scale-up of existing interventions. Vaccine characteristics explored include efficacy, duration of protection, uptake and cost. Results: Vaccines under all scenarios strongly reduce HIV incidence and AIDS-related deaths, with more efficacious vaccines of longer protection providing greatest benefit. Added to an optimized scale-up of existing interventions, a 60% efficacious and well implemented vaccine could avert up to 8.9 million infections by 2050, reducing annual new infections to 84,000 in LMICs. Added to a 50% scale-up of existing tools, the vaccine could avert up to 6 million new infections, reducing annual new infections to 367,000. Cost-effectiveness is highly sensitive to efficacy, cost, and duration of protection, but less sensitive to uptake. Added to an optimized scale-up of existing interventions the vaccine would be cost-effective at costs per regimen of less than $40 in lowincome countries. A vaccine added to 50% scale-up would be costeffective at costs per regimen below $80. Conclusions: With existing and other emerging treatment and prevention options, AIDS vaccines could be fundamental to achieving and sustaining the end of AIDS. These data provide strong evidence for sustained research and development toward efficacious AIDS vaccines at acceptable cost for incorporation into comprehensive programs. 394 HIV Research for Prevention 204 HIV R4P

395 Posters 48: Risk Perception P48.0 Significant Prevalence of Heterosexual Anal Sex among FSWs in India Highlights the Need for Specifically Tailored Interventions for HIV Prevention Mallika Alexander, Shweta Chidrawar, Sucheta Deshpande 2, Ramesh Paranjape 3 National AIDS Research Institute, Clinical Trial Unit, Pune, India, 2 National AIDS Research Institute, Pune, India, 3 National AIDS Research Institute, Director, Pune, India Thursday, 30 October P48.02 Knowledge and Perception on HIV Discordance, Transmission and Prevention among Couples in Durban, South Africa Mammekwa Mokgoro, William Kilembe 2, Hildah Shumba 2, Miriam Kamusoko, Tarylee Reddy 3, Elisabeth Dissen 2, Jonathan Davitte 4, Annie Mwaanga 2, Mark Brockman 5, Thumbi Ndung u, Susan Allen 4 HIV Pathogenesis Programme, Durban, South Africa, 2 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 3 Medical Research Council, Durban, South Africa, 4 Emory University School of Medicine, Atlanta, GA, United States, 5 Simon Fraser University, Burnaby, BC, Canada Background: Receptive anal sex increases HIV transmission risk by 20-fold per act as compared to receptive vaginal intercourse. Reports on Heterosexual Anal Sex (HAS) in Female Sex Workers (FSW), an important core population for HIV transmission are limited. HAS and its association with awareness, vulnerability, social empowerment and effect of focused intervention was studied in a large cross sectional survey among FSWs in India. Methods: Data from 809 FSWs from 9 districts of three high prevalence States of India from Round 2 Integrated Behavioural and Biological Assessment (IBBA) are presented. IBBA was aimed at studying the impact of Avahan, one of the largest HIV prevention programmes in India. Results: HAS was reported by 5.6%. FSWs who were literate (AOR.28 ( )) with good knowledge of STI symptoms (AOR.85( )) reported HAS. No other source of income (AOR.32(.0-.57)) and high volume of sex work in a week in terms of number of days (AOR.43(.6-.77)) and clients (AOR.25( )) were associated with HAS. Accessing STI treatment from Avahan clinic (AOR 2.0( )) and greater sense of empowerment (4.06( )) were positively correlated. Membership to Avahan Self Help Group (AOR 0.70( )) and STI testing at Avahan clinic (AOR 0.83( )) were protective. Conclusions: Ignorance of FSWs of HIV transmission potential through anal sex and hence need for specific interventions is highlighted. Higher report of anal sex by FSWs who accessed STI treatment emphasises need for doctors and nurses to openly discuss anal sex and treat anal STIs. Lifeskill training sessions aimed at empowering FSWs could be designed as a platform for safe anal sex messages as indicated by higher reporting of HAS by those with greater sense of empowerment. Focused service delivery and structural intervention through trained NGOs working with the core group will help in addressing HIV transmission through HAS. Findings also stress the need for rectal microbicide for women too. Background: Most incident HIV infection in sub-saharan Africa occurs in cohabiting discordant heterosexual couples. Though Couples Voluntary Counseling and Testing (CVCT) is an effective, well studied intervention in Africa, < 0% of couples have been jointly tested. Methods: As part of a 203 pilot project to expand CVCT in Durban, South Africa, a survey was conducted pre and post-cvct intervention to assess change in knowledge on HIV discordance, HIV transmission and prevention. Partners of a couple were interviewed separately using true/false responses to statements. The McNemar Chi -square test using STATA was applied to assess change in knowledge from baseline to post-cvct for each statement. Results: This study included 38 couples (636 individuals), all black South African; mostly Zulu ethnicity (86%), Christian (84%) and at least secondary level educated (76%). The mean age and range for men and women was 3 (7-59) and 29 (6-59) respectively. There was low level knowledge of possibility of HIV discordant results for a couple; only 95 individuals (3%) thought this was possible at pre CVCT compared to 578 (9%) at post-test. The survey also assessed knowledge on the benefit of CVCT; 208 (33%) thought there was at least one benefit at pre CVCT and this increased to 572 (90%) at post CVCT. Overall, there were statistically significant positive changes in knowledge on HIV transmission and prevention. In comparison between pre and post CVCT responses, most people thought that all HIV positive mothers give birth to babies with AIDS (63% and 58%; p=0.002). Most people also thought that Male circumcision does not protect HIV negative men against HIV (70% and 66%; p=0.05). Conclusions: Though the respondents had a positive attitude towards CVCT, the majority were initially unaware of the possibility of HIV discordance and there were misconceptions about HIV prevention and transmission. Future messages should target gaps in knowledge and provide information about CVCT services

396 Posters Posters 48: Risk Perception P48.03 P48.04 The Resonance Project: How Canadian Gay Men Understand, View and Incorporate Biomedical Knowledge of HIV into their Sexual Practices Marc-André LeBlanc, Ed Jackson 2, Barry Adam 3, San Patten 4, Len Tooley 2, James Wilton 2, Shayna Buhler 5, Greg Penney 6, Kim Thomas 7, Wayne Robert 8, Jody Jollimore 8, Joshun Dulai 8, Owen McEwen 9, Daniel Pugh 9, Robert Rousseau 0, Gabriel Girard 0, Alexandre Dumont-Blais 0 Resonance Project, Gatineau, QC, Canada, 2 CATIE, Toronto, ON, Canada, 3 University of Windsor, Windsor, ON, Canada, 4 San Patten & Associates, Halifax, NS, Canada, 5 Interagency Coalition on AIDS and Development, Ottawa, ON, Canada, 6 Canadian Public Health Association, Ottawa, ON, Canada, 7 Canadian AIDS Society, Ottawa, ON, Canada, 8 Health Initiative for Men, Vancouver, BC, Canada, 9 Gay Men s Sexual Health Alliance, Toronto, ON, Canada, 0 RÉZO, Montreal, QC, Canada Background: In the last 0 years, our biomedical knowledge of HIV prevention has grown tremendously and several new prevention tools are now at our disposal. Historically, gay men have been early adopters of risk reduction strategies, such as condoms. The Resonance Project is a 3-year community-based research project, funded by the Canadian Institutes of Health Research, seeking to understand how biomedical HIV knowledge is entering the discourses, prevention strategies and folk wisdom of gay men and their service providers. Methods: Using vignettes of typical online profiles and dating scenarios, we conducted 2 semi-structured focus groups with 86 gay men in Montreal, Toronto and Vancouver. The focus groups included 4 categories: ) gay men connected to the HIV sector (other than employment), 2) HIV + and - gay men in serodiscordant relationships, 3) HIV+ men who are sexually active, and 4) HIV- men who are sexually active and at high risk. We then conducted additional in-depth individual interviews with 0 of the gay men. Topics we explored included: seroadaptive behaviours, rapid and home-based testing, acute HIV infection, ARV-based prevention options, vaccines, and cures. The audio recordings were transcribed, coded, and analyzed using Interpretive Description (Thorne, 2008). Results: Gay men who participated reflected diverse attitudes, perspectives and backgrounds, based on ethnocultural origins, age, serostatus, relationship status, and city location. Depending on the extent to which the new biomedical knowledge had resonated with them, they demonstrated different levels of uptake of the information and resulting personal calculations of risk. Conclusions: Effective HIV prevention interventions need to enable gay men to develop their own risk management strategies in ways that resonate with their sexual lives. They must also take into account the ways in which gay men s biomedical knowledge of HIV can be at once sophisticated and incomplete, innovative and inaccurate, complementary and contradictory. Mobility Patterns and Concurrent Sexual Relationships among Fisher Folk along Lake Victoria, Kenya Stella Njuguna, Zachary Kwena 2, Everlyn Ombati, Margaret Mburu 2, Elizabeth Bukusi, Raphael Ondondo, Betty Njoroge Kenya Medical Research Institute, Nairobi, Kenya, 2 Kenya Medical Research Institute, Kisumu, Kenya Background: HIV prevalence in Kenya dropped from 7.4% in 2007 to 5.6% in 202. Despite this decrease, the HIV prevalence in Nyanza region remains the same, 5.%. This high prevalence may be driven by the highly migratory high risk fishing communities residing along the shores of Lake Victoria in Nyanza, with HIV prevalence at 25.6%. We examined the association between mobility patterns and perceived sexual practices of the fisher folk. Methods: Beach management units registers were used to identify and determine the number of fisher folk in every fish landing beach on the shores of Lake Victoria in Kenya. Probability proportionate to size sampling was used to draw a random sample of 2638 across 308 beaches for participation in a cross-sectional survey. Data was collected on sociodemographics, mobility patterns and reported sexual concurrency. Descriptive statistics and logistic regressions were used for data analysis. Results: In 203 mobility was mainly to Homabay and Siaya counties which are high HIV prevalent areas, ~36% and 3% respectively. Mobility was highest between April and December, 67 (33.3%) and 9 (23.7%) in Homa Bay and Siaya respectively. Approximately 36% suspected their partners had concurrent partners and 566(46.%) of the respondents reported that they were currently in a concurrent relationships. Mobility was significantly associated with reported sexual concurrency (OR % CI p = 0.009). After controlling for age, gender, education level, income, reported condom use, age of last born child, marital status, alcohol use and duration of relationship with the most recent sexual partner, mobility was significantly associated with concurrency (AOR.74 95% CI p=0.046). Conclusions: Mobility and concurrent sexual practices among fisher folk may contribute the sustained high HIV prevalence in Nyanza and calls for targeted HIV prevention and care services. 396 HIV Research for Prevention 204 HIV R4P

397 Posters 48: Risk Perception P48.05 Thursday, 30 October P48.06 How Canadian Service Providers Understand, and Incorporate, Biomedical Knowledge of HIV into their Prevention Work and their Own Sexual Practices San Patten,2, Ed Jackson 3, Barry Adam 4, Marc-Andre Leblanc 5, Len Tooley 3, James Wilton 3, Shayna Buhler 6, Kim Thomas 7, Greg Penney 8, Wayne Robert 9, Jody Jollimore 9, Joshun Dulai 9, Owen Mcewen 0, Daniel Pugh 0, Robert Rousseau, Gabriel Girard, Alexandre Dumont-Blais San Patten and Associates, Inc, Halifax, NS, Canada, 2 Resonance Project, Halifax, NS, Canada, 3 CATIE, Toronto, ON, Canada, 4 University of Windsor, Windsor, ON, Canada, 5 Resonance Project, Gatineau, QC, Canada, 6 Interagency Coalition on AIDS and Development, Ottawa, ON, Canada, 7 Canadian AIDS Society, Ottawa, ON, Canada, 8 Canadian Public Health Association, Ottawa, ON, Canada, 9 Health Initiative for Men, Vancouver, BC, Canada, 0 Gay Men s Sexual Health Alliance, Toronto, ON, Canada, REZO, Montreal, QC, Canada Background: In the last 0 years, our biomedical knowledge of HIV prevention has grown tremendously and several new prevention tools are now at our disposal. Historically, gay men have been early adopters of risk reduction strategies, such as condoms. The Resonance Project is a 3-year community-based research project, funded by the Canadian Institutes of Health Research, seeking to understand how biomedical HIV knowledge is entering the discourses, prevention strategies and folk wisdom of gay men and their service providers. Methods: We conducted 3 semi-structured focus groups with 22 service providers who work with gay men in Montreal, Toronto and Vancouver. This was followed by 20 one-on-one interviews with service providers, 0 of which were with service providers who identify as gay men. Topics we explored included: the challenges of conveying biomedical information to gay men; the organizational supports and constraints faced by service providers; and, the complex dual professional/personal position of service providers who are gay men. The audio recordings were transcribed, coded, and analyzed using Interpretive Description (Thorne, 2008). Results: Service providers reflected a wide variety of attitudes, level of understanding, degree of comfort and willingness to incorporate biomedical aspects of HIV prevention into: ) their HIV prevention messages and interventions; and 2) in the case of service providers who are gay men, their own sexual practices. Some of the latter service providers experienced challenges in reconciling their own sexual practices with the advice they give to gay men in their communities, vis-à-vis their emerging biomedical knowledge of HIV risk and prevention. Conclusions: Service providers play a critical role in implementing HIV prevention interventions that are reflective of gay men s lives and new biomedical understandings of HIV. Special attention is required to recognize and mobilize the dual position of service providers who are gay men in shaping prevention responses. Risk Disinhibition: The Effect of Microbicide Communication Materials on Future Prevention Behavior Elizabeth E. Tolley, Allison P. Pack, Sam Field 2, Elizabeth Ryan 3, Bockh Emily 4, Caroline Mackenzie 5, Alice Olawo 5, George Githuka 6 FHI 360, Social & Behavioral Health Sciences, Durham, NC, United States, 2 FHI 360, Quantitative Sciences, Durham, NC, United States, 3 FHI 360, Social Marketing and Communication, Washington, DC, United States, 4 FHI 360, Global Health, Population & Nutrition, Washington, DC, United States, 5 FHI 360, Country Office, Nairobi, Kenya, 6 National AIDS & STI Control Programme, Nairobi, Kenya Background: Vaginal gel use may provide a new HIV prevention tool for women who cannot negotiate condom use. Yet, current microbicide formulations are less effective than consistent condom use. Before introducing such options, it is essential to develop and test communication materials that encourage gel use in the absence of condom use, while promoting condoms-only or gel+condoms among those already using condoms to avoid reduced protection. In Kenya, we developed and assessed the effect of a minimum package of materials on preferences for future HIV prevention and possible risk disinhibition. Methods: We compared participants reports of ever condom use with a current/recent partner to future HIV prevention preferences (no product, gel-only, condoms-only, or gel+condom) among 746 sexuallyactive women and men in two Kenyan cities, who received informationonly, or viewed either HIV-framed or non-hiv framed awarenessraising materials (posters, TV and radio spots). We also assessed future prevention preferences among 99 young women and female sex workers (FSWs) pre- and post- participation in educational sessions using tailored flipcharts. Results: Risk disinhibition was low regardless of framing or material. After viewing awareness-raising materials, 80% of participants would maintain condom use, or increase their protection by adding gel to condom use or moving from no protection to gel, condoms or both. Married monogamous participants were significantly less likely than others to increase protection; 2% reporting past condom use preferred using no protection or gel-only. Preference for less protection after participating in educational sessions remained low (2% pre and post-test for FSWs) or decreased (from 7.5% to 2.5% in young women). Conclusions: Microbicide introduction must position use as possible, even desirable, by stable couples, while reinforcing the idea of gel as added protection for condom users or casual couples. Accomplishing these goals requires careful message development and testing

398 Posters Posters 48: Risk Perception P48.07 Conceptualizations of Uncertainty and Risk and Implications for Biomedical HIV Prevention Technologies in Sub-Saharan Africa: A Systematic Review Emily Warren, Pauline Paterson, Shelley Lees 2, Robyn Eakle 3, Jonathan Stadler 3, Heidi Larson London School of Hygiene and Tropical Medicine, Epidemiology and Population Health, London, United Kingdom, 2 London School of Hygiene and Tropical Medicine, Public Health and Policy, London, United Kingdom, 3 Wits Reproductive Health & HIV Institute, Hillbrow, South Africa Background: Two HIV pre-exposure prophylaxis (PrEP) trials, Voice and Fem PrEP, have recently been stopped due to lack of efficacy, often attributed to poor adherence. While oral PrEP was shown to be effective in other settings, its lack of efficacy, particularly for African women has prompted an urgent re-examination of assumption about HIV risk perception and the quotidian context in which sexual transmission of HIV occurs. Our aim is to understand how healthcare providers and individuals considered in the general or high-risk populations define and classify risk and navigate uncertainty in relation to sexual transmission of HIV, and identify how those conceptualizations are associated with the use biomedical prevention interventions. Methods: We are conducting a systematic review and meta-ethnography of qualitative findings, which allow us to keep the explanatory context intact. Six databases, Medline, Embase, PsychInfo, Africa Wide Info, CINAHL, and Global Health, were searched. To be included, papers employ qualitative methods to explore people s perception of risk, uncertainty, hope, trust, fear and/or decision making in relation to biomedical HIV prevention interventions. Because of feasibility concerns, we limited our included articles to only those that examined sexual transmission of HIV. Results: After duplicates were removed from our search results, two authors screened 8,826 articles. Screening is ongoing and more detailed results will be available before October. Conclusions: To the best of our knowledge, this is the first systematic review to explore different conceptions of risk and uncertainty in relation to the use of HIV biomedical technologies. This is especially important given the increased number of prevention interventions currently being researched. 398 HIV Research for Prevention 204 HIV R4P

399 Posters 49: Sexually Transmitted Infections P49.0 Socio-demographic and Behavioural Characteristics Associated with HSV-2 Seroprevalence in High Risk Women in KwaZulu- Natal, South Africa Nathlee S. Abbai, Handan Wand 2, Ramjee Gita,3 South African Medical Research Council, HIV Prevention Research Unit, Durban, South Africa, 2 The Kirby Institute, University of New South Wales, Sydney, Australia, 3 London School of Hygiene & Tropical Medicine, Department of Epidemiology and Population Health, London, United Kingdom Thursday, 30 October P49.02 Burden and Predictors of HIV /Hepatitis B Coinfection in Rural Uganda Clara Wekesa, Gershim Asiki, Ivan Kasamba, Rebecca N. Nsubuga, Robert Newton, Liz H. Young 2,3, Manjinder Sandhu 2,3, Alex Karabarinde, Anatoli Kamali MRC/UVRI, Uganda Research Unit on AIDS, Kampala, Uganda, 2 Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 3 University of Cambridge, Public Health and Primary Care, Cambridge, United Kingdom Background: The World Health Organization estimated that 536 million people aged 5-49 are infected with Herpes simplex virus type 2 (HSV-2), the causative agent of genital herpes. The aim of this study was to investigate the role of behavioral and demographic factors that contribute to the high HSV-2 sero-prevalence among women participating in a HIV prevention trial. Methods: The Methods for Improving Reproductive Health in Africa (MIRA) study assessed the effectiveness the latex diaphragm and lubricant gel on HIV prevention among women in South Africa and Zimbabwe. At screening an interviewer administered questionnaire on demographics and sexual behaviour was obtained. HSV-2 serum antibodies was detected using HerpeSelect TM2 ELISA IgG. Statistical analysis was performed using STATA release 2.0. Univariate and multivariate analyses were performed using logistic regression for both. Results: In this study, the prevalence of HSV-2 was estimated at 73% with 4% of the women also co-infected with HIV. In the multivariate analyses, older women (35 years and older) were more likely to be HSV-2 sero-positive as compared to younger women (OR: % CI: 2.7, 4.49 and OR:.82 95% CI:.49, 2.22) respectively. Women with a lower level of education were also considered to be more likely to be HSV2 positive (OR:.26 95% CI:.03,.53). Additionally, having > life-time sexual partners (OR: 2.48, 95%.92, 3.20) parity > (OR:.95 95% CI:.92, 3.20) and being HIV positive (OR: 6.3, 95% CI 5.06, 7.88) were all significantly associated with HSV-2 infection. Conclusions: The high sero-prevalence of HSV-2 in the studied population is of great public health importance since this high risk population could act as a reservoir for future infections particularly HIV transmission. The identification of risk factors for HSV-2 infection could aid in the development and implementation of personalized prevention messages for this infection. Background: Hepatitis B (HBV) and HIV have similar transmission patterns and thus HBV/HIV co-infection is common and this increases morbidity, mortality and risk of transmission. We estimated the burden of HBV/HIV co-infection and determined the associated factors in rural Uganda to inform the design of targeted preventive interventions. Methods: : In 20 a population based survey was conducted among residents of an established rural general population cohort (aged 3 years) in SW Uganda. Data on demographic characteristics, risky sexual behaviour (multiple and casual sex partners, condom use) were collected and all participants were tested for HIV (ELISA+ Western Blot), HBV (HBsAg) and liver function. Participants with confirmed infections were referred for care where viral loads, CD4 count (HIV+ only) were done. We determined the proportion of HBV/HIV co-infection and explored associated factors by logistic regression. Results: Of 8078 participants tested, 7% had HIV infection only; majority were female (66%) and the mean age was 38 years (±.5SD), 36% were on ART with a mean CD4 count of 367.6cell/mm 3 (±.8SD). The HBV prevalence was 4.6% (27/584) among those infected with HIV and 3.0% among HIV uninfected participants. Compared to those infected with HIV only, HBV/HIV co-infected participants were more likely to have had sex with a casual partner in the last 2 months (3.8%vs 4.4% p=0.04) and to have elevated liver transaminases (p=0.04). Sex, number of sexual partners, CD4 count, HIV RNA viral load and use of ART were not associated with co-infections. Conclusions: In this population we found high risk sexual behaviour as a predictor of HBV/HIV co-infection. The presence elevated liver enzymes may be a surrogate marker for acute hepatitis reflecting a higher risk of liver damage among co-infected participants. Preventive interventions that target both HIV and HBV are needed among high risk groups

400 Posters Posters 49: Sexually Transmitted Infections P49.03 Factors Associated with Sexually Transmitted Infections and HIV Risk in the Carraguard Phase 3 Trial Barbara A. Friedland, Doug Taylor 2, Khatija Ahmed 3, Thesla Palanee 4, Marlena Plagianos 5, Lauren Katzen 5 Population Council, HIV and AIDS Program, New York, NY, United States, 2 FHI 360, Durham, NC, United States, 3 Setshaba Research Centre, Soshanguve, South Africa, 4 Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 5 Population Council, New York, NY, United States P49.04 Patterns of Drug Use among People who Inject Drugs (PWID) and their Implications for Sexually Transmitted Infections in Northern Nigeria Desmond A. Iriaye, Ibrahim Suleiman, Otibho Obianwu, Sylvia Adebajo, George Eluwa, Jean Njab, Ayodeji Oginni, Babatunde Ahonsi Population Council, HIV/AIDS, Abuja, Nigeria Background: To describe the burden of sexually transmitted infections (STIs), correlates of prevalent infection, and associations between prevalent infection and HIV risk among South African women regularly tested and treated for curable STIs in an HIV prevention trial. Methods: We conducted a secondary analysis of data from 6004 women who participated in the Carraguard Phase 3 trial between 2004 and 2007 to further assess the burden of STIs and related risk factors. Women were treated for STIs based on positive test results or if clinically indicated, and were given referral letters for partners to access testing and treatment. We classified prevalent gonorrhea, chlamydia, trichomoniasis and syphilis in 6-month follow-up periods (up to 24 months). We used adjusted logistic regression and survival analysis to assess relationships between time in study, types of partnerships, previous STI status and other time-varying factors on prevalent and incident STIs. Results: STI prevalence fell significantly from baseline (25%) to Months -6 (2%) and Months 7-2 of follow-up (2%), after which prevalent infection with one or more STIs remained steady through Month 24. Younger age was associated with higher prevalence of gonorrhea and chlamydia (p< 0.0); the opposite relationship was observed for syphilis. Previous STI (except syphilis) and reports of new partners were positively associated with prevalence of STIs. Prevalent gonorrhea, chlamydia and trichomoniasis were also positively associated with risk of HIV seroconversion in adjusted models (HR=3.0, 2.7 and 2.4, respectively; P< 0.0). Conclusions: We observed a significant reduction in prevalence of curable STI over time in a trial with routine testing, treatment and partner referral. The overall STI burden remained substantial, however, emphasizing the ubiquitous nature of STIs in South Africa. Further reductions in STI prevalence would require including tests of cure and actively ensuring partner treatment, which may not be feasible in trial settings. Background: Drug use impairs judgment and leads to sexual risk behaviours. The prevalence of HIV among PWID in Kano (4.8%) and Kaduna (5.8%) respectively are higher than the national HIV prevalence among PWID (4.2%). However the national prevalence of sexually transmitted infections (STI) among PWID is about 6%. In this study we sought to determine drug use profile, prevalence and correlates of STIs among PWID in Northern Nigeria. Methods: Data of drug use, sexual and injecting risk behaviors were collected from structured risk assessment forms between May and October 203 and evaluated in a cross sectional study. Chi-square test was used to compare differences between categorical variables and logistic regression analysis was conducted to identify factors associated with patterns of drug use and sexual risk behaviour. Results: A total of 385 drug users (9% Females and 9% Males) with mean age of 30 years were interviewed. More than 94% injected drugs in the last 2 months. Mean age of drug use debut was5.4 years, while mean duration of drugs use was 5 years. Types of drugs injected included Heroine (30%), Cocaine (24%), Amphetamine (8%), Crack (0.4%) and Pentazocine (39.2%). Over 80% of heroin users and 72.3% of cocaine users, had unprotected sex with casual partner in the last 6 months. About 28% of drug users experienced one form of sexually transmitted infection in the last 6 months. Logistic regression analysis showed that amphetamine users were times more likely to have STI (Adjusted OR=.52, 95%CI= ) followed by Heroin users (Adjusted OR= 4.99, 95%CI= ) and Pentazocine users (Adjusted OR=2.86, 95%CI=.4-5.8). Conclusions: This study strongly suggested that drug use significantly predisposes PWID to sexual risk behaviours and STIs. In addition, exposure to STIs seems to significantly vary with the type of drugs used. Therefore, there is a need for appropriate HIV/STI prevention programing for PWID in Northern Nigeria 400 HIV Research for Prevention 204 HIV R4P

401 Posters 49: Sexually Transmitted Infections P49.05 Thursday, 30 October P49.06 Prevalent and Incident Gonorrhea and Chlamydia Infections among Oral Contraceptive and Depot Medroxyprogesterone Acetate Users in MTN- 003 (VOICE) Flavia Matovu Kiweewa, Lisa Noguchi 2, Barbra Richardson 3, Jen Balkus 3, Betty Kamira, Brenda Mirembe Gati, Clemensia Nakabiito, Gita Ramjee 4, Thesla Palanee 5, Gonasagrie Nair 6, Pearl Selepe 7, Ravindre Panchia 8, Patrick Ndase 9, Edward Livant 0, Ariane Van der Straten, Kailazarid Gomez 2, Devika Singh 9, Mary Glenn Fowler,2, Jeanna Piper 3, Zvavahera Mike Chirenje 4, Jeanne Marrazzo 9 MU-JHU Research Collaboration, Kampala, Uganda, 2 Johns Hopkins University, Baltimore, MD, United States, 3 SCHARP-FHCRC, Seattle, WA, United States, 4 MRC, Durban, South Africa, 5 Wits RHI, Johannesburg, South Africa, 6 CAPRISA, Durban, South Africa, 7 Aurum Institute, Klerksdorp, South Africa, 8 PHRU, Soweto, South Africa, 9 University of Washington, Seattle, WA, United States, 0 Magee-Womens Research Institute, Pittsburgh, PA, United States, RTI International, San Francisco, CA, United States, 2 FHI 360, Durham, NC, United States, 3 DAIDS/ NIAID/NIH, Bethesda, MD, United States, 4 UZ-UCSF, Harare, Zimbabwe Background: Oral contraceptive (OC) and depot medroxyprogesterone acetate (DMPA) use are common in areas with high sexually transmitted infection (STI) incidence. Recent STI increases risk of HIV acquisition and evidence suggests Chlamydia trachomatis (CT) risk may be higher among OC and DMPA users compared to users of no hormonal contraception. Data are limited regarding whether STI incidence differs between OC and DMPA users. We compared CT and Neisseria gonorrhoeae (NG) rates directly between OC and DMPA users. Methods: We analyzed prospective data from VOICE, a randomized trial of oral and topical HIV chemoprophylaxis in Uganda, Zimbabwe and South Africa. Analysis was restricted to 2,548 women who used OC (756) or DMPA (,792) exclusively during follow-up (f/u). Screening and treatment for CT and NG occurred at baseline (BL), annually, when clinically indicated, and at exit. We used site-stratified Andersen-Gill proportional hazards models to quantify CT and NG risk, excluding periods of no injectable or oral contraception and censoring at pregnancy and HIV seroconversion. Results: Compared to DMPA users, OC users were older (26.2 vs years, p< 0.00) and had lower BL prevalence of CT, although NG prevalence was similar (CT: 9.9% vs. 4.0%, p=0.0) (NG: 2.5% vs. 3.5%, p=0.23). Over 2,80 person-years (py) of f/u, 405 cases of CT (4.4/00 py) and 93 of NG (3.3/00 py) were detected. While incidence for both CT and NG was lower among OC compared to DMPA users (CT: 9.4 vs. 6.5/00 py; hazard ratio [HR] 0.59, 95% CI ) (NG:.5 vs. 4./00 py, HR: 0.37, 95% CI ), no significant differences were noted for CT or NG after adjusting for age, recent partner change, and other factors (CT adjusted HR [ahr]: 0.7, 95% CI ; NG ahr: 0.54, 95% CI ). Conclusions: Differences in CT and NG risk between OC and DMPA users may be related to demographic and behavioral factors. Providers should emphasize condoms and other STI prevention strategies for all women and their partners. Uptake of HBV Vaccination and Incident HBV Infection in Women of Reproductive Age and at Risk of HIV- Infection in the VOICE (MTN 003) Study Nyaradzo M. Mgodi, Cliff W. Kelly 2, Brenda Kakayi 3, James Y. Dai 2, Gonasagrie Nair 4, Thesla Palanee 5, Kailazarid Gomez-Feliciano 6, Jeanne Marrazzo 7, Jeanna Piper 8, Zvavahera Mike Chirenje University of Zimbabwe - University of California San Francisco Collaborative Research Programme, Harare, Zimbabwe, 2 SCHARP - FHCRC, Seattle, WA, United States, 3 Makerere University-Johns Hopkins University Research Collaboration, Kampala, Uganda, 4 University of KwaZulu Natal, CAPRISA, Durban, South Africa, 5 University of the Witwatersrand, Wits Reproductive Health and HIV Institute, Johannesburg, South Africa, 6 FHI 360, Durham, NC, United States, 7 University of Washington, Seattle, WA, United States, 8 DAIDS/NIAID/ NIH, Bethesda, MD, United States Background: In 99, the Global Advisory Group of the Expanded Programme on Immunization recommended that in areas of high HBV endemicity like Sub Saharan Africa (SSA), HBV vaccine be integrated into national immunization programs. Most SSA countries found this roll-out challenging given financial constraints and competing public health priorities. VOICE, a multi-site HIV PrEP trial, was conducted in SSA where high HBV and HIV prevalence rates exist. We aimed to determine the uptake of HBV vaccine and HBV incidence in VOICE. Methods: A total of 5029 women enrolled into VOICE. Prior HBV exposure was determined at screening (HbsAg, HBsAb). Screening exclusion criteria included HIV infection and HBV infection. Eligible HBsAg negative women were randomized equally to daily oral tenofovir disoproxil fumarate (TDF), TDF/emtricitabine, oral placebo, vaginal tenofovir or placebo gel; all were offered HBV vaccine if not immune (HBsAb negative) or counselling only if immune (HBsAb positive). We summarize HBV testing, vaccination uptake/completion and HBV incidence. Results: Three thousand six hundred and sixty seven (72.9%) women were not immune at enrolment. Of these, 3648 initiated the vaccination schedule at baseline or sometime during follow-up, yielding a total rate of HBV vaccination uptake of 99.5%. Two thousand nine hundred and eighty eight (8.5%) completed the 3-dose vaccination course as scheduled. A total of 330 (90.3%) completed > 3 doses, including cases where the vaccination course was interrupted and later restarted. Among the 3667 who were susceptible at baseline, 5(0.4%) acquired HBV during follow-up, with each HBV infection occurring after receipt of > 2 doses of vaccine. Of the 5, (20%) had HBV and HIV co-infection. Conclusions: Among VOICE participants, most were susceptible to HBV at enrolment, and uptake of vaccination was high. Despite HIV incidence of 5.7% during the study, the incidence of HBV in women at risk for HIV was low, presumably due to high uptake of effective vaccination. 40

402 Posters Posters 49: Sexually Transmitted Infections P49.07 Analysis of Griffithsin (GRFT) Binding by Proteins in Cervical Vaginal Lavage and the Influence of Vaginal Microflora Bernard J. Moncla,2, Lara K. Mahal 3, Brian M. Debo 2, Catherine Chappell 4, Katherine E. Bunge 4, Leslie Meyn 4, Jordan J. Szpara 2, Lisa C. Rohan 2,5, Sharon L. Hillier 2,4 University of Pittsburgh, Ob./Gyn and Reproductive Sciences, Pittsburgh, PA, United States, 2 Magee-Womens Research Institute, Microbiology, Pittsburgh, PA, United States, 3 New York University, Chemistry, New York, NY, United States, 4 University of Pittsburgh, Ob/Gyn and Reproductive Sciences, Pittsburgh, PA, United States, 5 University of Pittsburgh, Pharmacy, Pittsburgh, PA, United States P49.08 Prevalence of HIV, Sexual Transmitted Infections and High Risk Sexual Behaviors among Female Sex Workers in Kigali, Rwanda Jeannine Mukamuyango, Rosine Ingabire, Etienne Karita, Julien Nyombayire, Robertine Sinabamenye, Nuri Ahmed, Jean Nepo Nduwamungu, Sukaina Radjabari, Marydale Oppert 2, Matt Price 3, Frances Priddy 3, Pat Fast 3, Amanda Tichacek,2, Susan Allen,2 Rwanda Zambia HIV Research Group-Projet San Francisco, Kigali, Rwanda, 2 Emory University, School of Medicine, Department of Pathology and Laboratory Medicine, Atlanta, GA, United States, 3 IAVI, New York, NY, United States Background: Griffithsin (GRFT), an anti-hiv protein isolated from the red alga Griffithsia sp., is being developed as a microbicide. GRFT binds N-linked glycans on gp20 to prevent HIV entry. Earlier studies evaluating GRFT binding in cervicovaginal lavage (CVL) samples using lectin microarrays led us to investigate how GRFT interacts with glycoproteins in the vaginal microenvironment Methods: CVL was collected in PBS from 35 healthy asymptomatic reproductive aged women and stored at -70 C until used. Vaginal smears were evaluated using the Nugent criteria for bacterial vaginosis (BV). The samples were evaluated for GRFT binding in an ELISA using anti- GRFT primary antibody and a horseradish peroxidase labeled secondary antibody. The ELISA data were analyzed using one-way analysis of variance. GRT binding proteins in the CVLs were identified by SDS-PAGE and Western blotting. Results: GRFT bound at higher levels in the CVLs obtained from women having a Lactobacillus-predominant flora (Nugent score 0-3) or intermediate flora (Nugent 4-6) compared to BV flora (Nugent 7-0) in pg/ng protein: , , pg/ng protein, respectively, P value = < 0.00(Normal vs BV P< 0.00; Intermediate vs BV P=0.00). Western blots of normal CVL revealed five GRFT binding proteins at apparent molecular weights (in kda) of 85, 42, 20, 8 and 7, while 2/4 BV CVL demonstrated no GRFT binding proteins and two had binding to proteins with apparent MWs of 54 to>250. The CVL from women with BV had reduced levels of these GRFT binding proteins. Conclusions: GRFT binds to at least 5 proteins found in CVL, with different protein patterns in women with normal flora vs those with BV. The lower binding of GRFT in the CVL of women with BV suggests a protective function for GRFT binding proteins. It also suggests that a GRFT based microbicide may be bound to endogenous vaginal proteins reducing availability to compete with the HIV gp20 protein in women with normal flora. Background: Female sex workers (FSW) are at high risk of transmitting/ acquiring HIV and other Sexually Transmitted Infections (STIs). Despite widespread education, counselling, and condom distribution interventions among FSW, unprotected sexual intercourse persists. This study aimed to assess the level of HIV prevalence among FSWs who consulted VCT services at Projet San Francisco in Kigali, Rwanda. Methods: From September 202-March 204, Projet San Francisco provided HIV voluntary counseling and testing, STI screening and treatment as well as Long Acting and Reversible Contraceptive (LARC) methods to FSW invited from known hot spots of commercial sex activity in Kigali. HIV negative women are offered enrollment into a prospective study on HIV incidence, while HIV positive women are referred to government health centers for care and treatment. Results: During the study, 605 FSW received services at PSF, of whom 263 (43%) tested HIV negative. The mean age was 3 years with a range of 6-5. HIV prevalence rose with age from 3% in < 24 year olds to 56% in those aged 25-29, 58% in year olds and 70% among those >35. At baseline, 264 (44%) FSW were diagnosed with at least one STI. The most commonly diagnosed STI was Syphilis (5 cases; 25%), followed by Trichomoniasis (84 cases; 4%), Pelvic Inflammatory diseases (33 cases; 5%), and Gonorrhea (5 cases; %). 2 women were treated for more than one STI. The average number of sex clients per month was 44 (range: 0-500), the median was 20 clients per month and the proportion of FSW who reported consistent condom use for all sex acts in the last month was 75%. Conclusions: FSW in Kigali have a very high prevalence of HIV and other STI. Increasing HIV testing and counseling is especially needed at an early age to decrease risk of new HIV infections. Specific interventions for this high-risk population are needed to limit the spread of the HIV/ AIDS epidemic. 402 HIV Research for Prevention 204 HIV R4P

403 Posters 49: Sexually Transmitted Infections P49.09 Sexually Transmitted Infections: Co-infections and Link to Variations in HIV Prevalence and Disease Management Outcomes across Regions in Kenya Thursday, 30 October P49.0 Prevalence and Determinants of Hepatitis B Virus (HBV) Infection in a Cohort of HIV- Discordant Couples in Western Kenya: Implications for HIV Care Joseph Mwangi,2, Joyceline Kinyua, Nancy Lagat, Raphael Lihana, HIV Coinfection and Molecular Study Group Kenya Medical Research Institute, Nairobi, Kenya, 2 Insititute for Tropical Medicine and Infectious Disease, Jomo Kenyatta University of Science and Technology, Nairobi, Kenya Background: Whereas many STIs have gained significance over the years and especially with regard to HIV and AIDS, Herpes Simples Virus 2 and Syphilis are some of the most important. Herpes simplex virus 2 (HSV-2) and syphilis are important etiologic agents of sexually transmitted infections. In this era of HIV and AIDS, the two infections have become a major public health concern due to their association with HIV acquisition and pathogenesis by driving viral replication and facilitating transmission. In this study testing for the two infections was carried out to determine levels of infections and possible associations including CD4 counts since differences in HIV infections have been indicated across regions in Kenya. Methods: Three sites across 3 regions in Kenya were included in the study where 7 HIV positive participants were included. CD4 count, HSV and Syphilis status were tested using FacsCalibur and serological tests respectively. Results: Among the study participants, 66.9% were female and 33.% Male. Test results indicated 67.5% of the participants were HSV Positive while 32.5% were Negative with females having a higher prevalence. Syphilis sero-status was at 2.4% and 87.6% positive and negative respectively.cd4 count ranged from to 700 counts /ul of blood with differences noted between genders, test results status as well as between sites/regions. Conclusions: Higher prevalence of STIs, including HSV and syphilis has potential to contribute to increased HIV transmission and could reverse the gains being made with treatment for preventions especially in HIV discordant relations and increased incident noted among certain population groups and regions in Kenya. Dismas C.O. Oketch, Eunice Kaguiri, Nereo Murgor 2, Cosmas Apaka 3, Paul Ayuo 4, Edwin Were 5, Kenneth Fife 6 Moi University School of Medicine, Partners in Prevention, Eldoret, Kenya, 2 Moi University School of Nursing, Eldoret, Kenya, 3 Moi University College of Health Sciences, Center of Excellence for Cardiovascular & Respiratory Diseases, Eldoret, Kenya, 4 Moi University School of Medicine, Department of Internal Medicine, Eldoret, Kenya, 5 Moi University School of Medicine, Department of Reproductive Health, Eldoret, Kenya, 6 Indiana University School of Medicine, Department of Medicine & Infectious Diseases, Indianapolis, IN, United States Background: Hepatitis B is a major global health problem with over 350 million people suffering from chronic HBV infection. Infection with HBV negatively impacts the outcome of HIV infection and yet there are no national clinical guidelines for diagnosis, treatment and care for HBV- HIV co-infection. This study sought to determine the prevalence and determinants of HBV infection in a cohort of HIV- discordant couples in Western Kenya. Methods: A cross sectional study of healthy heterosexual HIV- discordant couples from Western Kenya referred for possible recruitment into the Partners PrEP Study was conducted between September 2008 and October 200. All participants were screened for hepatitis B surface antigen (HBsAg) and socio-demographic data were collected. Descriptive statistics were used to determine frequencies while the association between HBsAg and the independent variables were evaluated using logistic regression. Results: Data on 834 adults aged 8-64 are presented. HBsAg was positive in 77 [4.2%] individuals. Men were 40% less likely to be infected compared to women [OR 0.6; 95% CI ; p value ]. Those from rural areas were almost 2 times more likely to be infected than those from urban areas [OR.92; 95% CI ; p value 0.008]. Hepatitis B prevalence did not differ by HIV status [HIV positive 3.9% vs negative 4.5% [OR=.5; 95% CI of p value = 0.56]. There was also no significant correlation between hepatitis B and participants age, alcohol intake, CD4 counts, income or level of education. Immunity to hepatitis B (as measured by the presence of antibody to HBsAg) was identified in 25.6% [469] with the majority [73%] of the participants being in their 3 rd and 4 th decades. Conclusions: Whereas HBV vaccination should be scaled up for both adult men and women irrespective of their HIV status, more effort should be directed towards adult rural women of child bearing age especially those below 25 years

404 Posters Posters 49: Sexually Transmitted Infections P49. Burden of Sexually Transmitted Infections Among High Risk Women Screened for a Phase III Microbicide Trial in Southwestern Uganda Martin Onyango, Andrew Abaasa, Anita Kabarambi, Faith Naddunga, Sylvia Kusemererwa, Neliette Van Neikerk 2, Annalene Nel 2, Anatoli Kamali Medical Research Council/ Uganda Virus Research Institute, Uganda Research Unit on AIDS, Entebbe, Uganda, 2 International Partnership for Microbicides, Paarl, South Africa P49.2 Number Needed to Screen for Chlamydial and Gonococcal Infection among Asymptomatic Men Who Have Sex with Men in Bangkok, Thailand, Sarika Pattanasin, Punneeporn Wasinrapee, Jaray Tongtoyai, Wannee Chonwattana, Anuwat Sriporn, Pikunchai Luechai, Wichuda Sukwicha, Anupong Chitwarakorn 2, Timothy H. Holtz,3, Marcel Curlin,3 Thailand-MoPH U.S. CDC Collaboration, Nonthaburi, Thailand, 2 Ministry of Public Health, Department of Diseases Control, Nonthaburi, Thailand, 3 Division of HIV/AIDS Prevention, U.S. Centers for Disease Control and Prevention, Atlanta, GA, United States Background: Sexually Transmitted Infections (STIs) facilitate transmission and acquisition of HIV infection. Screening for STIs is recommended in HIV microbicide trials and untreated STIs delay enrolment. We determined the burden of STIs among high risk women screened for a Phase III microbicide trial in Southwestern Uganda. Methods: The Medical Research Council/Uganda Virus Research Institute (MRC/UVRI) in collaboration with International Partnership for Microbicides (IPM) is evaluating safety and efficacy of a dapivirine vaginal ring among healthy HIV-negative high risk women in a multicentre Phase III trial. These women were identified from bars, lodges, restaurants, and hair salons in townships and fishing communities within 50 km from the research centre. High risk was defined by presence of STIs, self-reported unprotected sex with multiple sex partners, and frequent use of recreational drugs (marijuana, alcohol) in the past 3 months. All consenting female volunteers were screened for STIs regardless of clinical symptoms. Cervico-vaginal samples were taken for T. vaginalis (OSOM Rapid test), C. trachomatis/n. gonorrhoeae (Roche PCR test), and blood samples taken for syphilis (RPR/TPHA). Treatment was as per the Uganda National guidelines for syndromic management and CDC STI treatment guidelines. The proportion of STIs among the women screened was determined. Results: Of the 236 women screened between August 203 and March 204, the mean age was 28 years (range 8 to 42 years). About half (, 47%) had T. vaginalis, 23 (0%) C. Trachomatis, 20 (8%) N.gonorrhoeae and 2 (5%) syphilis (RPR and TPHA positive). A total of 43 (8.2%) had at least 2 or more STIs. Conclusions: There is a high burden of STIs among high risk women. Though such populations are considered suitable for vaginal microbicide efficacy trials, the high STI burden impacts greatly on trial enrolment. There is a need for regular STI screening, partner identification and treatment, and condom promotion in this population. Background: Asymptomatic infection caused by Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) are common among men who have sex with men (MSM). The U.S. Centers for Disease Control and Prevention (CDC) recommends screening sexually active MSM at least annually for CT and NG infection. We calculated the number of asymptomatic MSM needed to screen (NNS) to detect one MSM with CT and/or NG, stratified by sexual role. Methods: Between 2006 and200, we enrolled Thai MSM, age 8 years, from the Bangkok metropolitan area. Participants completed sexual behavior questions using audio computer-assisted self-interviews, and underwent physical examination, and rectal, urethral, and pharyngeal screening for CT and NG using nucleic acid amplification testing (NAAT). We calculated NNS as the reciprocal of the proportion of asymptomatic MSM with unrecognized infection detected by NAAT. Results: Of,744 participants enrolled, 593 (9%) had no symptoms or signs of CT/NG at baseline. We detected CT infection in 26 (4%), NG infection in 99 (6%), and CT/NG co-infection in 40 (2.5%). The overall NNS to detect CT and/or NG at any sites was 5 (95% Confidence Interval [CI] 5-6). Among insertive-only MSM (n=285), the chance of detecting urethral CT infection (NNS, 95% CI 8-9) was higher than detecting urethral NG infection (NNS 7, 95% CI , p< 0.05). Among versatile MSM (n=284), the NNS for infection at any site was 7 (95% CI 6-8) for CT, and 4 (95% CI 2-8) for NG. Among receptive only MSM (n=287), the NNS for infection at rectal was 6 (95% CI 5-9). Conclusions: Asymptomatic CT and NG infection in this population is high. Given that the NNS in asymptomatic sexually active MSM is low, annual screening using NAAT for all sites can diminish progression of diseases and disrupt transmission. In settings where screening all specimen sites is not available, the rectal swab test should be a priority. 404 HIV Research for Prevention 204 HIV R4P

405 Posters 49: Sexually Transmitted Infections P49.3 ABSTRACT WITHDRAWN Thursday, 30 October P49.4 HIV- + /HSV-2 + Co-infection Is Associated with Persistence of CD4 + and DC-SIGN + Antigen Presenting Cells at the Mucosa Independent of HSV Recurrences Gloria C. Preza, Marla J. Keller 2, Maria T. Ochoa, Betsy Herold 3 University of Southern California, School of Medicine, Los Angeles, CA, United States, 2 Albert Einstein College of Medicine, Pediatric Infectious Diseases, Bronx, NY, United States, 3 Albert Einstein College of Medicine, Pedriatic Infectious Diseases, Bronx, NY, United States Background: Prior studies demonstrate that HSV-2 seropositivity is associated with higher HIV plasma viral loads & increased genital HIV shedding. We hypothesize that in HIV + /HSV-2 + women, genital HSV outbreaks will be associated with an increase in antigen presenting cells (APCs), which may facilitate recruitment of HIV target cells & contribute to replication & transmission in HIV + /HSV-2 + women. Methods: A 2-week study was conducted among HIV + /HSV-2 +, HIV + / HSV-2 -, HIV - /HSV-2 + & HIV - /HSV-2 - women to examine the impact of HSV- 2 reactivation on mucosal immunity. Vaginal biopsies were collected at baseline &from a lesion & contralateral uninvolved site during a clinical recurrence. Sections from 4 HIV + /HSV-2 + & 4 HIV - /HSV-2 + women were evaluated for APCs & results quantified by in situ IHC imaging analysis. Results: At baseline, co-infected women had significantly more CD209 + (DC-SIGN) & CD4 + (monocytes/macrophages) cells compared to HIV + / HSV-2 - &HIV - /HSV-2 + women. After an outbreak, there was an increase in these APCs in HIV - women at the lesional but not the contralateral site (p<.02). However, the response in co-infected women was blunted. For example, CD209 + expression increased from ± 78.6 cells/mm 2 at baseline to only 32.3 ± 99.4 cells/mm 2 in lesions, but increased in lesions from HIV - women from 25.3 ± 92 cells/mm 2 at baseline to ± 2 cells/mm 2 (p<.004). Similarly, there was little or no increase in CD4 + cells in lesions in co-infected women whereas CD4 increased in HIV - women from 54.5± 8.9 to 29.6 ± 9.7 cells/mm 2 (p<.04). Conclusions: Co-infection is associated with an increase in CD209 + & CD4 + APCs in biopsies independent of HSV disease. During clinical HSV recurrences, there is an increase in APCs at the lesional but not the contralateral site, which is more pronounced in HIV - women. The largest APCs observed were CD4 + monocytes & CD209 + macrophages. These findings provide a potential mechanism for the increased risk of HIV transmission in the setting of HSV-2 co-infection

406 Posters Posters 50: Tenofovir Gel: Acceptability and Adherence P50.0 P50.02 FACTS 00: Characteristics of Participants Enrolled in a Phase III Randomised Controlled Trial of Tenofovir Gel for Prevention of HIV- and HSV-2 Sinead Delany-Moretlwe, Emilee Smith 2, Landon Myer 2, Khatija Ahmed 3, Busi Nkala 4, Rebone Maboa 5, Cynthia Gama 6, Sydney Sibiya 7, Modulakgotla Sebe 8, William Brumskine 9, Linda-Gail Bekker 0, Maposhane Nchabeleng, Carl Lombard 2, Glenda Gray 4,2, Helen Rees, FACTS 00 Study Group University of the Witwatersrand, Wits Reproductive Health & HIV Institute, Johannesburg, South Africa, 2 University of Cape Town, Centre for Infectious Diseases Epidemiology & Research, Cape Town, South Africa, 3 Setshaba Research Centre, Soshanguve, South Africa, 4 University of the Witwatersrand, Perinatal HIV Research Unit, Soweto, South Africa, 5 University of the Witwatersrand, Wits Reproductive Health and HIV Institute, Johannesburg, South Africa, 6 University of the Witwatersrand, MATCH, Durban, South Africa, 7 Qhakaza Mbokodo Research Centre, Ladysmith, South Africa, 8 Aurum Institute, Tembisa, South Africa, 9 Aurum Institute, Rustenburg, South Africa, 0 University of Cape Town, Desmond Tutu HIV Centre, Cape Town, South Africa, Medunsa, Medunsa Clinical Research Unit (MeCRU), Ga-Rankuwa, South Africa, 2 Medical Research Council of South Africa, Cape Town, South Africa Background: FACTS 00 is a phase III licensure trial to assess the safety and effectiveness of tenofovir % gel applied intravaginally before and after sex in preventing HIV- and HSV-2 in women in South Africa. We report on the baseline characteristics of the trial population. Methods: Participants were screened for eligibility for the trial at nine sites across South Africa from October 20 until March 204. Women aged 8-30 years, who were HIV- negative, sexually active, using a reliable modern contraceptive, with no travel plans, no recent history of enrolment in another HIV prevention trial, and without evidence of renal or pelvic disease, raised liver enzymes, or pathological bone fractures, who consented to all study procedures, were considered eligible for the trial. Participants were enrolled irrespective of HSV-2 status at enrolment. Results: 3846 participants were screened and 2059 were enrolled in the trial. The main reasons for exclusion were HIV- positive, not willing or able to consent to all procedures, and evidence of proteinuria or glycosuria. At the time of enrolment, the mean age of participants was 23 years, and the majority of participants were unmarried (88%), lived with parents (62%), had completed school (43%), were unemployed and earned < R000 in the past three months (73%). Most participants reported having only one regular partner (89%) and a median of 9 (IQR 5-6) sex acts in the past 28 days. The overall prevalence of HSV-2 at enrolment was 42%. Conclusions: The baseline characteristics of FACTS 00 enrolled trial population reflect those most at risk for HIV infection in South Africa, and in need of new HIV prevention technologies. Meta-analysis of the Effectiveness of Tenofovir Gel in Preventing HIV Infections Anneke Grobler CAPRISA, Congella Durban, South Africa Background: Three clinical trials have been conducted to test whether % Tenofovir gel prevents heterosexual transmission of HIV. The CAPRISA 004 study found a reduction of 39% and the VOICE study found a reduction of 4.7% in HIV incidence comparing the Tenofovir gel arm to the placebo arm. The FACTS study is still under way and results are expected early next year. The objective is to estimate the combined efficacy of tenofovir gel using all three studies. Methods: A meta-analysis was done of the CAPRISA and VOICE results. Additional meta-analyses also included hypothetical results of the FACTS study. The Cochran-Mantel-Haenszel common relative risk was calculated for each meta-analysis. The CAPRISA and FACTS studies had coitally dependent dosing (the so-called BAT24 strategy), while the VOICE study had daily dosing. Results: A meta-analysis of the gel arms in VOICE and CAPRISA 004, estimates that Tenofovir gel provides 24% protection (95%CI: 2-4%,p-value=0.03). If the results of the FACTS study was similar to the VOICE study, the meta-analysis estimates 2% protection (95%CI: 2-36%,p-value= 0.03) and if it was similar to CAPRISA 004 the metaanalysis estimates 28% protection (95%CI:-42%,p-value=0.003). If the FACTS study showed a null-result (the same number of infections in the Tenofovir gel and placebo arms) then the meta-analysis estimates that Tenofovir gel provides 8% protection (--34%,p-value=0.068). In all instances the test for heterogeneity did not detect heterogeneity between the studies. Conclusions: If the results of the FACTS study lie between the results of the CAPRISA and the VOICE studies, i.e. provides between 4.7% and 39% protection from HIV infection, one expects the combined efficacy estimate of the three studies to lie between 2% and 28%, with a p-value smaller than This would indicate that Tenofovir gel is efficacious in preventing HV infection, when all available efficacy trials are combined. 406 HIV Research for Prevention 204 HIV R4P

407 Posters 50: Tenofovir Gel: Acceptability and Adherence Thursday, 30 October P50.03 P50.04 Tenofovir Gel as a Component of the HIV Prevention Mix: Perspectives from Participants, Partners and Community Men in KwaZulu-Natal, South Africa Kathleen M. MacQueen, Sarah Dlamini 2, Brian Perry, Eunice Okumu, Steve Sortijas, Chitra Singh 2, Diantha Pillay, Sharon Watson FHI 360, Durham, NC, United States, 2 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa Background: Many questions exist with regard to how % tenofovir gel could be integrated with existing HIV prevention methods, if it is approved. We explored perceptions of women and men about the gel relative to other prevention methods in the context of CAPRISA 008, an ongoing open-label follow-on trial of tenofovir gel that enrolled uninfected CAPRISA 004 trial participants. Methods: In-depth interviews (n=63) and focus groups (n=8) were held with CAPRISA 008 participants; male partners of 3 women who fully disclosed trial participation and gel use were also interviewed. Four focus groups were held with community men who were not partners of women in the CAPRISA 008 trial. Perspectives on HIV prevention options including tenofovir gel were assessed using an applied thematic analysis approach. Results: Male condoms and male circumcision were perceived to be the most effective means of HIV prevention by both female and male respondents; however, the difficulty of consistent condom use often negated perceived efficacy while male circumcision faced cultural aversion and was viewed as mainly protecting men. Few respondents reported use of female condoms; most were averse to the thought of using them. Tenofovir gel was generally viewed as an effective means of HIV prevention by both women and men, though there was concern that it is still not licensed by the government and that it is not 00% effective. Use of tenofovir gel was particularly considered beneficial for women if their partners refuse to wear condoms and was described as easy to use relative to other HIV prevention methods. Few participants reported cases of condom migration. Some men noted that their partner s use of tenofovir gel provided risk reduction benefits to them as well. Conclusions: Both men and women in KwaZulu-Natal, South Africa perceive tenofovir gel as offering the potential to address limitations in the current HIV prevention method mix. Tenofovir Gel Acceptability and Adherence among Pregnant Women in the United States Elizabeth T. Montgomery, Lisa Noguchi 2, James Dai 3, Jason Pan 3, Joseph Biggio 4, Karen Isaacs 5, Heather Watts 6, Jill Schwartz 7, Jeanna Piper 8, Richard Beigi 2 RTI International, Womens Global Health Imperative, Los Angeles, CA, United States, 2 University of Pittsburgh Medical Center and Magee-Women Research Institute, Pittsburgh, PA, United States, 3 SCHARP - FHCRC, Seattle, WA, United States, 4 University of Alabama at Birmingham, Birmingham, AL, United States, 5 FHI 360, Durham, NC, United States, 6 NIH/NICHD, Bethesda, MD, United States, 7 CONRAD/ EVMS, Arlington, VA, United States, 8 NIH/NIAID, Bethesda, MD, United States Background: The MTN-008 trial, the first multi-dose study of a microbicide gel (2: randomized to tenofovir % or HEC placebo gel) during pregnancy, included daily pregnant participant gel insertion at home for 5 days. Because pregnancy may pose unique challenges to consistent gel use and acceptability these factors were assessed. Methods: Participants completed a web-based computer-assisted selfinterview (CASI) at Days 0 and 6 about gel attitudes and behaviors. At Day 6 trained research staff conducted a short interviewer-administered questionnaire with both structured and open-ended questions. Frequencies of quantitative data were tabulated in SAS; qualitative data were manually summarized. Results: CASI data include 96 participants in both active and placebo gel arms. Self-reported adherence was high with 88% reporting daily gel use and the majority reporting no difficulty with daily use. The most common reason for nonuse was forgetting. Participants reported generally neutral perceptions of gel characteristics. Two-thirds thought the gel was runny, many complained of bothersome gel leakage and several cited this reason for not inserting a full dose. A small number of women (6-8%) reported pain, other physical discomfort, or mental discomfort associated with the process of applicator insertion, and fewer than 5% reported the same in regards to the gel itself. Although the majority were not worried the gel would cause problems for their pregnancy or babies, one-quarter to one-third felt somewhat worried about these issues. Ten percent of women said they would not use the gel in the future when not pregnant, while 38% would prefer to use gel only before sex and 32% had no preference between daily and precoital gel use. Conclusions: Self-reported adherence to gel for 5 days at home was high. Gel was generally acceptable, but many complained of a runny consistency and leakage. There were no frequent or strong concerns about the effects of the gel on the pregnancy/ fetus reported

408 Posters Posters 50: Tenofovir Gel: Acceptability and Adherence P50.05 P50.06 LB Group Adherence Support (GAS) - Experiences from CAPRISA 008 Chitra Singh, Nelisiwe Ngcobo, Sarah Dlamini, Carl Montague, Fanele Ntombela, Nabeela Warrasally, Kershia Sunjeevan, Quarraisha Abdool Karim, Leila Mansoor Centre for AIDS Programme of Research in South Africa (CAPRISA), Durban, South Africa Background: Adherence is key to ensuring effectiveness of ARV based microbicides. Adherence patterns may differ in a placebo controlled trial where efficacy is uncertain compared to an open-label trial of a product of known efficacy and also with increasing duration of follow-up. Participant experiences and challenges in adhering to using % tenofovir gel in CAPRISA 008 was assessed as part of ongoing adherence support. Methods: Five group adherence support (GAS) sessions were conducted between January and February 204 by a trained staff member. Participants who were enrolled in the study for >2 months volunteered to take part in these GAS sessions that comprised groups of 6-0 participants and lasted minutes. Detailed notes transcribed during these sessions were reviewed to identify common themes and differences within and across sessions. Results: A total of forty six women aged between years participated in these GAS sessions. Common issues emerged regarding factors affecting adherence to gel use over time. Initial confidence about product use started to be eroded with time as they learnt of participants becoming infected with HIV. In four out of five sessions it was reported that partner disclosure of gel use enhanced adherence. In three out of five sessions participants reported that unplanned coitus, partner complaints of vaginal wetness and leakage of gel, influenced adherence over time. In order to address these barriers to adherence participants suggested the following: inserting the gel earlier in the day to reduce wetness at the time of coitus especially if they have not disclosed gel use to their partner and storage of gel in convenient and multiple locations (i.e. cosmetic bag/pencil case) for easy access particularly when unplanned coitus takes place. Conclusions: These pilot GAS sessions identified both barriers and facilitators to product adherence. Participants welcomed suggestions made by peers to enhance their product adherence levels. A Phase Evaluation of the Rectal Safety, Acceptability, Pharmacokinetics, and Pharmacodynamics of Three Formulations of Tenofovir % Gel Ian McGowan, Kathy Duffill 2, Alexiy Nikiforov 2, Charlene Dezzutti, Nicola Richardson-Harman 3, Kaleab Abebe, Amy Herrick 4, Mark Marzinke 5, Ross Cranston, Lisa Rohan 6, Craig Hendrix 5, Hirot Hiruy 5, Julie Elliott 7, Christine Mauck 8, Peter Anton 7, and the CHARM Program Research Team University of Pittsburgh School of Medicine, Pittsburgh, PA, United States, 2 Magee Womens Research Institute, Pittsburgh, PA, United States, 3 Alpha StatConsult LLC, Damascus, MD, United States, 4 University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA, United States, 5 Johns Hopkins University School of Medicine, Baltimore, MD, United States, 6 University of Pittsburgh School of Pharmacy, Pittsburgh, PA, United States, 7 University of California at Los Angeles, Los Angeles, CA, United States, 8 CONRAD, Arlington, VA, United States Background: CHARM-0 was undertaken to characterize the rectal safety, acceptability, pharmacokinetics (PK), and pharmacodynamics (PD) of three formulations of tenofovir (TFV) % gel: a vaginal formulation (VF), a reduced glycerin vaginal formulation (RGVF), and a rectal formulation (RF) with osmolalities of 3,, 846, and 479 mosmol/kg respectively. The VF gel was previously used in the CAPRISA 004 and VOICE vaginal microbicide Phase 2B trial. The VF gel was used in the RMP-02/MTN-006 Phase rectal safety study. The RGVF gel was used in the MTN-007 Phase rectal microbicide trial and is currently being used in the MTN-07 Phase 2 rectal microbicide trial. Methods: Participants received 4 ml of study product in a blinded, randomized, crossover design: (i) 6 daily doses of a HEC placebo followed by a final single dose of the VF gel, (ii) 7 daily doses of the RGVF gel, and (iii) 7 daily doses of the RF gel. Safety, acceptability, compartmental PK, and explant PD were monitored throughout the trial. Results: Thirteen participants were enrolled into the CHARM-0 study. All three gels were found to be safe and acceptable. Median rectal tissue homogenate TFV-diphosphate (DP) concentration was significantly greater with the RF (0.3 ng/mg) versus the VF (below the limit of quantification) gel. Median mucosal mononuclear cell TFV-DP was significantly greater with the RF (36 fmol/0 6 cells) versus VF (9 fmol/0 6 cells) gel (p 0.05). Use of each gel in vivo was associated with significant inhibition of ex vivo colorectal explant HIV infection. There was also a significant relationship between tissue levels of TFV/ TFV diphosphate (DP) and inhibition of explant infection. Conclusions: All three formulations were safe and acceptable. There was a trend towards higher tissue levels of TFV associated with exposure to the RF gel but and all three gels were associated with significant inhibition of explant infection. The RGVF or the RF gel could be advanced into later stage development as a rectal microbicide. 408 HIV Research for Prevention 204 HIV R4P

409 Posters 50: Tenofovir Gel: Acceptability and Adherence P50.07 LB Introducing Microbicides: Estimated Cost of QI Approach and Tenofovir Gel Delivery with Family Planning Services in the CAPRISA 008 Trial Thursday, 30 October Rick Homan, Sarah Dlamini 2, Leila Mansoor 2, Imraan Valodia 3, Steve Sortijas, Sharon Watson, Kristine Torjesen FHI 360, Durham, NC, United States, 2 CAPRISA/University of KwaZulu Natal, Durban, South Africa, 3 University of Witwatersrand, Johannesburg, South Africa Background: The CAPRISA 008 implementation trial of % tenofovir (TFV) gel randomizes participants to receive TFV gel with family planning (FP) services in a primary health care (PHC) or research clinic setting in South Africa. The CAPRISA 06 ancillary study estimated the cost of using a quality improvement (QI) approach to strengthen FP services and integrate TFV gel provision in the PHC clinic at Vulindlela between June 202 and January 204. Methods: QI activity data were extracted from 86 documents and estimates of eligible client volume, gel uptake and FP visit recall were developed from PHC and CAPRISA service statistics. Unit costs were applied to resources using KwaZulu-Natal Department of Health (DOH) data and other sources. Financial labor costs (additional payment required) were separated from opportunity labor costs (time spent during routine work). Results: The value of resources required at the district level per PHC clinic for the first year of gel delivery is estimated at 9,946 ZAR ($89,500) and for the QI approach is 90,94 ZAR ($8,660). Recurring annual financial cost for gel delivery is estimated at 895,264 ZAR ($87,850) per clinic. Major financial cost at the district level is for TFV gel procurement with minor labor costs for staff training. The value of resources required at the PHC level per clinic for the first year of gel delivery is 309,587 ZAR ($30,400) and for the QI approach is 26,96 ZAR ($2,460). For gel delivery, recurring annual financial cost is 282,74 ZAR ($22,740) per clinic for additional labor, HIV test kits and other supplies. For the QI approach, recurring annual financial cost is,545 ZAR ($52) with opportunity costs estimated at.25 nurse days/month and 2.25 data clerk days/month. Conclusions: The resources required for introducing TFV gel are dominated by cost of the product, as well as additional labor to provide services at PHC clinics. Compared to the costs of TFV gel, introducing a QI approach to support gel delivery imposes minimal cost to the DOH

410 Posters Posters 5: Therapeutic Vaccine and Viral Latency P5.0 Partial Protection by Vaccination Does Not Prevent Chronic Neuro-inflammation in an SIV Model P5.02 In vitro Latent Virus Reactivation Potential of a Novel Triterpenoid Compound and Two Cytostatic Gold(III) Complexes Debbie H. Ferguson, Claire Ham, Atze Das 2, Ben Berkhout 2, Andrea Meiser 3, Steve Patterson 3, Neil Berry, Neil Almond National Institute of Biological Standards and Control, Virology, Potters Bar, United Kingdom, 2 Academic Medical Centre, University of Amsterdam, Amsterdam, Netherlands, 3 Imperial College London, London, United Kingdom Pascaline Fonteh, Petrina Kapewangolo 2, Debra Meyer University of Pretoria, Department of Biochemistry, Pretoria, South Africa, 2 University of Namibia, Department of Chemistry and Biochemistry, Windhoek, Namibia Background: With antiretroviral therapy (ART), HIV is now a chronic viral infection. However, suppression of viral load does not prevent chronic inflammation and the development of co-morbidities such as HIV-associated neurocognitive impairment. Using a non-accelerated ART-free SIV model that exhibits reproducibly controlled viral kinetics, we have demonstrated early neuroinvasion and chronic progressive neuropathology occurs even when peripheral viremia is well controlled. Here we investigated the impact on chronic neuroinflammation of either switching off a conditional live SIV or modifying early viremia. Methods: Immunohistochemical analysis of macaque brain sections was performed. Firstly, groups of macaques were infected with SIVrtTA, a doxycycline (DOX) dependent, conditional-live SIV. We compared groups on daily DOX treatment for 40 weeks with those where DOX was removed at week 3. Secondly, we compared groups of macaques immunised with an SIVgag based vaccine regimen. Vaccination did not reduce set point viral loads, however, it did significantly blunt primary viremia compared with naïve challenge controls. Results: Study. Chronic neuroinflammatory responses were detected in all SIVrtTA infected macaques even 37 weeks after DOX treatment was removed. Indeed, there was evidence astrocytosis and microgliosis were lower in macaques receiving the anti-inflammatory doxycycline throughout the study. Study 2. Suppression of primary viremia reduced neuroinflammation compared with that seen in naïve challenge controls. Conclusions: In conclusion, primary viremia alone is sufficient to initiate chronic neuroinflammation. Blunting primary viremia modulates subsequent neuropathology but does not prevent it. The neuroinflammatory legacy of acute infection even with subsequent potent viral control implies patients may suffer significant co-morbidities despite improved ART or even following eradication of HIV by functional cure. Background: Virus elimination by combining latency activators and highly active anti-retroviral therapy (HAART) is one way of eradicating HIV that is currently under investigation. Here, we present a comparative study of the virus reactivation potential of a novel triterpenoid plant product () derived from Ocimum labiatum to that of two synthetic cytostatic bis(thiosemicarbazonate) gold(iii) complexes (2 and 3). Methods: Viral reactivation was quantified by measuring p24 antigen production from a chronic infection and viral latency model of HIV (U cells). To exclude toxicity effects in the reactivation study, a tetrazolium dye was used to determine cytotoxicity. The mechanism of viral reactivation was determined by measuring the effect of, 2 and 3 on protein kinase C (PKC) and histone deacetalyse enzyme activity in an ELISA and a fluorometric assay respectively. A cytometric bead array kit was used to quantify the endogenous production of TNF-α. Results: The CC 50 of, 2 and 3 on the U cells was found to be 8.2 ±0.µg/mL, 0.53±0.2 µm and.0±0.4 µm respectively. Non-toxic concentrations (exhibiting >90% viability) of, 2, and 3 significantly reactivated virus (p< 0.05) by 3.5, 2.7 and 2.3 fold from U cells. Compound and complexes 2 and 3 minimally activated PKC levels but did not inhibit HDAC indicating that the reactivation mechanism does not involve these enzymes. A notable observation was the increase in TNF-α by up to 7 fold, possibly as a result of generalised immune activation by these compounds. Conclusions: The upregulation of TNF-α by, 2 and 3 is considered an off target effect which is part of a non-specific mechanism of viral reactivation. These findings suggest that the novel triterpene and the cytostatic bis(thiosemicabazonate) gold(iii) complexes could potentially be incorporated in drug cocktails aimed at activation/elimination of latent HIV but should be modified using techniques such as nanotechnology to improve selectivity and exclude off target effects. 40 HIV Research for Prevention 204 HIV R4P

411 Posters 5: Therapeutic Vaccine and Viral Latency Thursday, 30 October P5.03 P5.04 Reactivation of Latent HIV- by Bryostatin- in the Presence of Antiretroviral Drugs Marta Martínez-Bonet, Maria Isabel Clemente, Susana Alvarez, Laura Diaz, Dolores García-Alonso, Ana Lopez, Santiago Moreno 2, Eduardo Muñoz 3, Mariangeles Munoz-Fernandez Hospital General Universitario Gregorio Marañón, IISGM, Networking Research Center of Bioengineering, Madrid, Spain, 2 Hospital Universitario Ramón y Cajal, and IRYCIS, Madrid, Spain, 3 Instituto Maimónides de Investigación Biomédica de Córdoba, Universidad de Córdoba, Cordoba, Spain Background: Despite the fact that the antiretroviral therapy (ART) successfully controls HIV- in most infected patients, virus latency established early upon infection impedes HIV- eradication. Bryostatin- (BRYO) in vitro inhibits HIV--infection of CD4+T lymphocytes and, at the same time, reactivates HIV- through PKC- NF-kB pathway. The potential effects of BRYO in combination with antiretrovirals, such as maraviroc (MVC) and Atripla (ATP) is needed to be determined in vitro prior to design clinical trials. Methods: Jurkat-LTRGFP-R5 cell line and two latent and reactivatable HIV--infected J89GFP lymphocytic or THP89GFP monocytic clonal cell lines were used as latency models. Results: BRYO reactivates HIV- reaching levels >80% in THP89GFP cells, even in combination with MVC or ATP. Moreover, when MVC or ATP pre-treated reporter TZM-bl cells were co-cultured with BRYO treated with THP89GFP cells, new infections of reactivated X4/R5-HIV were inhibited 50% or 80%, respectively. Remarkably, when antiretrovirals were combined with BRYO, the combinations maintained the antiviral effect of the drug with the maximum rate of inhibition on its own. Also, BRYO-mediated downregulation of surface CD4 and CXCR4 in primary blood mononuclear cells (PBMC) was not affected when it was used along with the other antirretrovirals and no hiperactivation or high proliferation effects were observed in these PBMC. Significantly, BRYO was also tested ex vivo for HIV- induction in CD4+ T lymphocytes isolated from HIV- infected patients receiving ART and was found to exhibit potent viral induction activity. Conclusions: This work is the first to demonstrate that antiretrovirals combinations with BRYO do not interfere in the BRYO activity neither the BRY combination with antiretrovirals interferes in the antiviral activity of the antiretrovirals. Thus, we propose BRYO to purge the viral reservoirs, and this treatment should be combined with present ART. Bryostatin Activates HIV- Latent Expression in Human Astrocytes through a PKC and NFkB-Dependent Mechanism Laura Diaz, Marta Martinez-Bonet, Javier Sanchez-Rodriguez, Alejandra Fernández-Pineda, Esther Alonso, Jos Luis Jiménez, Eduardo Muñoz 2, Santiago Moreno 3, Susana Alvarez, Mª Ángeles Muñoz-Fernández Hospital General Universitario Gregorio Marañón, IISGM, Networking Research Center of Bioengineering, Madrid, Spain, 2 Instituto Maimónides de Investigación Biomédica de Córdoba, Universidad de Córdoba, Cordoba, Spain, 3 Hospital Universitario Ramón y Cajal, and IRYCIS, Madrid, Spain Background: Previous studies have suggested that the central nervous system (CNS) is an important reservoir for HIV and that proviral DNA and productive infections can be detected in this region. In particular, the brain represents a sanctuary for HIV- latency because the provirus can persist in this area due to the variable and poor penetration of antiretrovirals (ARV). Despite an undetectable viral load in patients treated with potent ARV, current therapies are unable to purge the virus from these latent reservoirs. Combinatory strategies to eliminate HIV- reservoirs using selective activators of viral expression could lead to a decline in HIV- levels in reservoirs that would be sufficient to efficiently control the infection. Methods: Human primary astrocytes (NHA) isolated from the cerebrums of 5-month-old human foetuses and astrocytic cell line U-87 were used in all experiments. Results: To broaden the inhibitory range and effectiveness of current ARV, the potential of bryostatin was investigated as a latent HIV- activator. Bryostatin reactivates latent viral infection in NHA and in U-87 via activation of protein kinase C (PKC)-alpha and -delta because the PKC inhibitors rottlerin and GF09203X abrogated the bryostatin effect. We did not find any alteration in cell proliferation. Moreover, bryostatin strongly stimulated LTR transcription by activating the transcription factor NF-κB. The effect of bryostatin could be especially important in a cellular reservoir such as astrocytes because it may be a beneficial adjunct to the treatment of HIV-infection. Conclusions: Bryostatin can directly increase HIV- LTR activity in human astrocytes in vitro via the PKC pathway and an NF-κB-dependent mechanism. Thus, it is plausible that HIV--infected astrocyte cells exposed to bryostatin may contribute to HIV- latency activation and will provide a foundation for future novel HIV--purging strategies from tissue reservoirs such as the CNS. 4

412 Posters Posters 52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV P52.0 P52.02 Couples Voluntary HIV Counseling and Testing (CVCT) followed by Treatment as Prevention (TasP) for Discordant Couples: The Impact of Each Step Susan Allen, William Kilembe 2, Mubiana Inambao 3, Rachel Parker 4, Tyronza Sharkey 2, Naeemah Munir 3, Linda Kimaru 3, Sam Scherber 2, Kristin Wall 5,6, Amanda Tichacek 4, Bellington Vwalika 2, Eric Hunter 7, Joseph Mulenga 8, Elwyn Chomba 9 Rwanda Zambia HIV Research Group-Emory University, Pathology & Laboratory Medicine, School of Medicine, Atlanta, GA, United States, 2 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Lusaka, Zambia, 3 Rwanda Zambia HIV Research Group, Zambia Emory HIV Research Project, Ndola, Zambia, 4 Rwanda Zambia HIV Research Group, Department of Pathology & Laboratory Medicine, SOM, Emory University, Atlanta, GA, United States, 5 Emory University School of Public Health, Epidemiology, Atlanta, GA, United States, 6 Rwanda Zambia HIV Research Group, Lusaka, Zambia, 7 Emory University School of Medicine, Department of Pathology & Laboratory Medicine, Atlanta, GA, United States, 8 National Blood Transfusion Services, Lusaka, Zambia, 9 Ministry of Community Medicine, Maternal & Child Health, Permanent Secretary, Lusaka, Zambia Background: Most new adult HIV infections in Africa are acquired from steady partners and WHO recommends that couples be tested and counseled together as an HIV prevention strategy. ART reduces transmission from the HIV+ partner in discordant couples. Methods: CVCT programs established in 70 Zambian government clinics include follow-up testing for discordant and concordant negative (M-F-) couples. We assess seroincidence in discordant couples, confirm linkage through sequencing, and compare rates before (antigen positive at CVCT or seroconverting within 6 weeks of CVCT) and after CVCT and on or off ART. We also compare the overall number of seroconversions in discordant and M-F- couples, and compare the cost of averting one HIV infection through CVCT vs. TasP. Results: 48,839 couples were tested, of whom 7,69 were discordant (3,229 or 8.3% on ART at the time of CVCT) and 09,677 were concordant negative (M-F-). Of 2 seroconversions, 62 occurred in discordant (5.4/00PY, 95% CI ) and 50 in M-F- (0.98/00PY, 95% CI ) couples. Among discordant couples, excluding infections acquired from outside partners, the rate before CVCT was 8.9/00PY (6.5-.8) compared with 2.3/00PY after CVCT (95% CI The overall transmission rate in discordant couples not on ART was 8.6/00PY (95% CI ), compared with 3.7 (95% CI ) when the infected partner was on ART. Among M-F- couples, incidence was.4/00py (95%CI.0-.9) before and 0.2/00PY (95% CI ) after CVCT. Using program costs and rates, the cost of averting one infection with CVCT was <$500, compared with an annual cost of >$0,000 for TasP in discordant couples. Conclusions: CVCT prompted a 74% reduction in new infections among discordant couples and a 9% reduction in M-F- couples. Among discordant couples TasP was associated with a 58% reduction. The combination of CVCT + TasP in discordant couples is ideal, but CVCT has significant impact even when TasP is not available, and at 5% of the cost of TasP. The Impact of Biological Monitoring by CD4 and Viral Load in Guiding the Decision to Initiate ART Therapy in HIV- Infected People Georgia Elna Ambada Ndzengué CIRCB, Yaoundé, Cameroon Background: Helper CD4 T cell count is utilized in the CDC categorization of HIV/AIDS disease progression and in guiding the decision to initiate antiretroviral treatment (ART) in many low income countries. However, in disease intense region exclusion of people from ART by CD4 values might not reflect the reality. Methods: From 20 to 203, HIV disease progression was monitored in a cohort of treatment -naïve HIV- infected people by CD4 count and plasmatic VL. Eligibility for treatment was assessed according to the WHO 200 guidelines. Results: Amongst 249 participants analyzed 40 % were eligible to treatment during the first visit when both helper CD4 count (257±70) and HIV- VL (5.2±0.79) was taken into consideration. However, when CD4 count alone was considered we recorded just 32% eligibility to treatment implying that 7.63% of eligible participants are left out. During the course of three years a further 5.3 % of the remaining participants became eligible to therapy by VL (5.5±0.77) and CD4 count (275±7) with 2 % being eligible by CD4 counts. With regards to the 200 WHO recommendations well over 49 % of participants were eligible to treatment during the course of three years. Conclusions: When the recent WHO 203 recommendations for the initiation of HAART are considered well over 40% of the remaining members of the cohort should be eligible to treatment. This suggests that close to 90 % of treatment naïve HIV- infected people are eligible within three years by this new recommendation. Thus a significant number of people would be eligible to treatment within a short period of time implying that the test and treat strategy should be envisaged in Cameroon. 42 HIV Research for Prevention 204 HIV R4P

413 Thursday, 30 October Posters 52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV P52.03 P52.04 Treatment as Prevention (TasP) in MSM and Transgender Women: Successful Detection of Acute/Recent Infection and Linkage to Care in Lima, Peru Ann Duerr, Javier Lama 2, Hugo Sanchez 3, Robinson Cabello 4, Jorge Sanchez 2 Fred Hutchinson Cancer Research Center, Seattle, WA, United States, 2 IMPACTA, Lima, Peru, 3 Epicentro, Lima, Peru, 4 Via Libre, Lima, Peru Background: The overall objective of this study is to improve the efficacy of Treatment as Prevention (TasP) strategies by increasing detection of acute and recent HIV infection. This research is being conducted in men who have sex with men (MSM) and transgender women (TW) in Lima, Peru, where knowledge about the symptoms of acute HIV infection and its high infectivity is low. Methods: We will enroll 800 HIV-uninfected MSM and TW at risk for HIV infection (STI in the past 6 months, self-identify as a sex worker, no condom use at last anal intercourse, anal intercourse with >5 partners in past 6 months, sex with an HIV-infected partner) into an acute infection cohort. Cohort participants are tested for HIV every month using a pointof-care third-generation EIA; negative specimens are pooled and tested for HIV RNA twice daily by real-time PCR. Participants who test positive for HIV RNA are contacted the next business day by peer counselors for linkage to the study site. Results: 2,027 MSM/TW were screened between July, 203 and April 30, 204. HIV-infected MSM/TW (5%) were referred to care and HIV-negative MSM/TW were enrolled into the cohort. In this period, we detected 27 acute HIV infections (HIV seronegative, HIV RNA positive). Thirty-eight participants had recent infections (seropositive: 7 had tested HIV negative in the past month and 3 within the past 3 months). Nine HIV+ participants did not continue in the study: one elected to be treated at a non-study clinic, two had psychiatric problems that precluded enrollment, 5 refused to participate due to logistics and other reasons, and we were unable to re-contact. Slightly over half of the HIV-infected participants were linked to care within 7 days of HIV diagnosis. Conclusions: Detection of acute and recent HIV infection and rapid linkage to care is possible. Successful linkage of such individuals to HIV care including ART can maximize TasP intervention and individual clinical benefits. This approach may be important for future cure strategies. Geographic Utilization of Gift Cards Used for Financial Incentives to Encourage Viral Suppression: Findings from HPTN 065 Theresa Gamble, Padraig Corcoran 2, Jill Stanton, Phaedrea Watkins, Elizabeth Greene, Jennifer Farrior, Richard Elion 3, Susan Amenichi-Enahoro 4, Wafaa El-Sadr 5,6, for the HPTN 065 Study Team FHI 360, Durham, NC, United States, 2 Massachusetts Institute of Technology, Cambridge, MA, United States, 3 Whitman-Walker Health, Washington, DC, United States, 4 Morris Heights Health Center, Bronx, NY, United States, 5 Columbia University Mailman School of Public Health, ICAP, New York, NY, United States, 6 Harlem Hospital, New York, NY, United States Background: HPTN 065 (TLC-Plus) evaluated the feasibility and effectiveness of providing quarterly $70 financial incentive gift cards to HIV infected patients on antiretroviral therapy who maintained viral suppression (HIV RNA< 400 copies/ml). A total of 39,359 FI gift cards were dispensed over 2 years by 9 HIV care sites in the Bronx, NY and Washington, DC. Methods: Data for each gift card disbursed included dispensing site, transaction amount, and location of transaction (zip code). Cards never used and transactions without valid zip codes were excluded. ZIP Code Tabulation Areas were used to map the location of transactions by venue (Bronx/DC) and dispensing site. Python programing and Microsoft Excel 200 were used for all analysis and visualization. For transactions that occurred outside of the local jurisdictions of DC and the Bronx, a random sample of transactions was examined to identify transactions made in person versus by internet/phone. Results: Of 78,529 transactions for gift cards distributed in the Bronx, 3,6(4.6%) transactions occurred outside of NY, linked to,852 unique gift cards of the 23,268 distributed. Of 62,022 transactions for gift cards distributed in DC, 3,928 (6.3%) occurred outside of DC, Maryland (MD) and Virginia (VA), linked to,987 unique gift cards of the 6,09 distributed. Transactions occurred in all 50 US states, Puerto Rico, the Virgin Islands and international locations. Of gift cards distributed in the Bronx but used outside of NY state, 62% were used in person and 74% of the gift cards distributed in DC, but used outside of DC, MD and VA, were used in person. Conclusions: Gift cards distributed in this study were primarily used locally; however, about 5% of transactions were outside the local jurisdictions and mostly in person. These data suggest that HIV infected individuals in the Bronx and DC travel throughout the US and beyond; thus, research is needed to understand their migration/travel patterns and the implications for interventions using ART for prevention. 43

414 Posters Posters 52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV P52.05 HIV Ascertainment through Repeat Homebased Testing in the Context of a Treatment as Prevention Trial (ANRS 2249 TasP) in Rural South Africa P52.06 Frequency of Achieving Viral Suppression among HIV-infected Persons in Serodiscordant Partnerships Initiating Antiretroviral Therapy Joseph Larmarange,2, Éric Balestre 3, Joanna Orne-Gliemann 3, Collins Iwuji 2, Nonhlanhla Okesola 2, Marie-Louise Newell 4, François Dabis 3, France Lert 5, TasP ANRS 2249 Group IRD, CEPED (UMR 96 Paris Descartes Ined IRD), Paris, France, 2 University of KwaZulu Natal, Africa Centre for Health and Population Studies, Mtubatuba, South Africa, 3 Inserm, ISPED (Unité 897 Inserm Université Bordeaux Segalen), Bordeaux, France, 4 University of Southampton, Faculty of Medicine, Faculty of Social and Human Sciences, Southampton, United Kingdom, 5 Inserm, CESP (Unité 08), Villejuif, France Background: The ANRS 2249 TasP cluster-randomised trial evaluates whether HIV testing of all members of a community, followed by immediate antiretroviral treatment (ART) for infected people, will prevent onward sexual transmission and reduce HIV incidence at population level. Ascertaining the HIV status of a high proportion of the population regularly and repeatedly is key to the success of any universal test and treat strategy, as the first step of the HIV cascade. Methods: Between March 202 and March 204, we implemented three six-monthly rounds of home-based HIV counselling and testing in ten local communities (clusters). At each home visit, individual questionnaires were administered and a rapid HIV test offered to all trial participants. We report early results on rates of HIV ascertainment, defined as undergoing a rapid HIV test or HIV-positive self-report. Results: Of 2,9 eligible individuals (resident in the trial area and 6 years), 0,007 were successfully contacted at least once. At first contact, HIV status was ascertained for 7,628 (76.2% [95% CI: ]) individuals. At second contact, among the 5,885 individuals contacted a second time, HIV status was ascertained for 2,829 (85.0% [95% CI: ]) of the 3,328 tested negative at first contact and for 543 (45.7% [95% CI: ]) of the,88 who refused a rapid test at first contact. Overall, HIV ascertainment rate was 89.0% (5,239/5,885 [95% CI: ]) among trial participants contacted twice. Conclusions: Repeat home-based HIV testing is acceptable and feasible in this rural area. Socio-demographic characteristics, behaviours, attitudes, household characteristics and experience of HIV infection and ART in the household will be explored for their association with HIV ascertainment uptake. This will inform whether this intervention reaches the individuals at higher risk in a rural South African region. Andrew Mujugira, Connie Celum, Allan Ronald 2, Nelly Mugo,3, Jared Baeten University of Washington, Seattle, WA, United States, 2 University of Manitoba, Winnipeg, MB, Canada, 3 Kenya Medical Research Institute, Nairobi, Kenya Background: Antiretroviral therapy (ART) markedly reduces the risk of HIV transmission in serodiscordant partnerships. The concentration of HIV RNA in plasma is the principal measure of ART adherence and is also the prime determinant of HIV transmission risk. Methods: We used data from a prospective study of 928 HIV-infected persons from Kenya and Uganda who had a known heterosexual HIVuninfected partner, to assess viral suppression after ART initiation. Plasma was archived 6-monthly and later tested for HIV RNA quantity. Kaplan-Meier methods, and Cox regression models, were used to identify independent predictors of viral suppression. Results: The median age was 35 years (interquartile range [IQR] 29, 4), and 502 (54%) were women. The median CD4 count and HIV RNA plasma concentration prior to ART start were 270 cells/µl (IQR 23, 370) and 4.34 log 0 copies/ml (IQR 3.65, 4.85), respectively. The cumulative probabilities of achieving viral suppression at 6 and 2 months were 82.8% and 90.8% for HIV RNA concentration < 400 copies/ml. Female gender (adjusted hazard ratio [AHR].9; 95% CI:.02,.39; p=0.03) predicted viral suppression. Older age (AHR.04 per-5 year increase; 95% CI: 0.99,.08; p=0.03), higher education (AHR.4; 95% CI: 0.99,.3; p=0.07 for >7 years), and higher CD4 count (AHR.6; 95% CI: 0.97,.38 for >200 compared with < 200 cells/µl) were of borderline significance. Conclusions: Three-quarters of HIV-infected persons with known HIVuninfected partners achieved viral suppression within 6 months of ART initiation. These results support World Health Organization guidance recommending ART initiation for persons with known HIV-uninfected partners, which could be optimized by viral load monitoring to identify those needing adherence support. 44 HIV Research for Prevention 204 HIV R4P

415 Thursday, 30 October Posters 52: Treatment as Prevention: Initiation, Adherence, and Monitoring on ARV P52.07 The Cascade of HIV Care: The Tool for Assessing Treatment as Prevention in Russian Federation P52.08 Patient Retention in Antiretroviral Therapy Programs up to Three Years on Treatment in Uganda, : A Retrospective Review Anastasia V. Pokrovskaya, Vadim V. Pokrovsky, Natalia N. Ladnaya, Oleg G. Yurin, Karl Emerole 2 Federal Budget Institution of Science Central Scientific Research Institute of Epidemiology of Rospotrebnadzor, Russian Federal AIDS Centre, Moscow, Russian Federation, 2 Peoples Friendship University of Russia, Department of Infectious Diseases, Moscow, Russian Federation Background: Success of treatment as prevention (TasP) depends on effective engagement of HIV-positive persons in care. The objective of this study was to estimate the cascade of HIV care and to identify gaps that impede realization of TasP in Russia. Methods: We defined 7 steps in the cascade of care framework: HIV infected (estimation data), HIV diagnosed, Linked to HIV care, Retained in HIV care, Need ART, On ART, Viral suppressed (VL <,000 copies/ml during 2 month ART). Information was extracted from the Federal AIDS Centre database (period 989 year - 3st of December 202) and also from the National monitoring forms of Rospotrebnadzor. Results: 6,858 HIV diagnosed Russian residents were alive by the end of 202, which consisted 47% of the estimated,300,63 people living with HIV. Among the HIV diagnosed patients, 470,892 (77%) were linked to care while 428,279 (70%) were retained. Of 29,87 (2% of HIV diagnosed) patients who were eligible for ART, 25,623 (97%) were on treatment and 02,383 (8%) of them had viral suppression. However, only 7% of HIV diagnosed patients acheived viral suppression which is necessary to prevent viral transmission. The most significant leakages of patients were on steps: HIV infected HIV diagnosed (loss - 53%), HIV diagnosed Linked to care (-23%) and Retained in care Need ART (-70%). Conclusions: The stages of HIV diagnosis and estimation of ART eligibility were the most vulnerable to leakage. Additional efforts are needed to encourage HIV testing to reduce the number of undiagnosed cases. Also earlier initiation and enlarging the recommendations to commence ART among patients retained to care is one of the key aims to optimize treatment as HIV prevention initiative in Russian Federation. Livingstone Ssali, Charles Ngobi 2, Bernard Michael Etukoit 2, Aggrey Egessa 3, Celestine Bakanda 4, Jonathan Wangisi 5, Stephen Okoboi 5 The AIDS Support Organisation,TASO, Monitoring and Evaluation, Kampala, Uganda, 2 The AIDS Support Organisation,TASO, Programme Management, Kampala, Uganda, 3 The AIDS Support Organisatio,TASO, Programme Management, Kampala, Uganda, 4 The AIDS Support Organisation,TASO, Planning and Strategic Information, Kampala, Uganda, 5 The AIDS Support Organisation,TASO, Research, Kampala, Uganda Background: Uganda is considered a success story in HIV and AIDS programming, but the extent to which adolescents have been retained in care still not well understood. Retention in care is important for positive clinical outcomes. The major of objective of the study was to assess the socio-demographic factors associated with retention in care, treatment and support programs among adolescents. Methods: We identified cohort of patients within TASO Management Information System, who were initiated on Antiretroviral Therapy in We conducted a retrospective review of data of adolescents aged 0-9 years who received ART services at TASO service centers from 2006 to 2009 Using Cox proportional hazard model, Hazard ratios (HR), and Adjusted Hazard ratios were computed. Confidence Intervals and appropriate p-values were computed to control for confounding. We estimated the overall retention level at 36 months after ART initiation including patients who died, lost to follow-up, transferred patients. Results: Female 390(63%), Male 227 (37%), age disaggregation 0-4 years 344(56%), 5-9 years 273(44%). In multivariate analyses, male clients had reduced probabilities to attrition adjusted hazard ratio (AHR) 0.79 (95% CI: 0.64 to 0.98), (p=0.040) compared to female. Adolescents who received ART at the health facility had increased risk to attrition, adjusted hazard ratio (AHR).3 (95%CI:.05 to.6), (p = 0.03) compared to those who received services at the Community drug distribution points (CDDPs). Adolescents with higher CD4 at ART initiation (cells/ml) CD cells/ml AHR.36 (95%CI:.05 to.77)) and >=250 cells/ml, AHR.33 (95%CI:.02 to.75), (p=0.036) had a high risk to attrition compared to adolescents with lower CD4 Conclusions: The study indicated that using Community Drug Distribution Points (CDDP) is an effective way to improve patient retention amongst adolescents receiving ART in a low-income setting. 45

416 Posters Posters 53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora P53.0 P53.02 Male Condom Functionality in the Presence of a Vaginal Ring Mildie Leuvennink, Neliette Van Niekerk, Terri Walsh 2, Ron Frezieres 2, Annalene Nel International Partnership for Microbicides, Clinical Affairs, Paarl, South Africa, 2 California Family Health Council, Los Angeles, CA, United States Background: IPM is evaluating the safety and efficacy of a dapivirine vaginal ring for the prevention of male to female HIV- transmission. Once the safety and efficacy of the ring have been proven and it is available for general use, women will be counselled to use male condoms together with the ring for maximum protection against HIV- infection. The compatibility of male condoms with the vaginal ring was evaluated. Methods: The total clinical failure rate of male condoms in the presence and absence of a placebo ring was assessed in an openlabel, randomized, crossover non-inferiority trial. Seventy healthy, monogamous, heterosexual couples, 8-45 years (females) or 8-55 years (males) were enrolled, and used 4 condoms when the female was wearing the ring and 4 condoms when the female was not wearing the ring. The total clinical failure rate was defined as the number of condoms that slipped off the penis or broke during intercourse, divided by the number of condoms used. The safety, tolerability and acceptability of male condoms in the presence of the ring were assessed. Results: 275 condoms were used with and without the ring. The total clinical failure rate was 2.2% with the ring and 4.0% without the ring. The difference with ring-without ring was -.9% (95% CI: -5.3%;.5%). The upper bound of the CI was less than the pre-defined non-inferiority margin of 3%. One male urogenital discomfort (severe burning) occurred during condom use without the ring, due to the condom being too tight. One participant reported a product-related adverse event (AE) of mild penile pain during the period of vaginal ring use. No serious AEs were reported; no AE led to trial discontinuation. More male than female participants felt the vaginal ring during intercourse, but the majority reported not being bothered by the ring. Conclusions: Male condom use was safe and well tolerated with vaginal ring use. The presence of the ring did not negatively affect the total clinical failure rate of male condoms or condom use experience. Baseline Characteristics of HIV-negative Women Enrolled into the Ring Study - A Clinical Trial of the Dapivirine Vaginal Ring for HIV- Prevention Annalene Nel, Neliette Van Niekerk, Allison Carter, Hannelie Carstens, Michelle Isaacs, Val Kidd, John Steytler, Linda-Gail Bekker 2, Cynthia Gama 3, Katherine Gill 2, Anatoli Kamali 4, Philip Kotze 5, Cheryl Louw 6, Nokuthula Miti 7, Hugo Tempelman 8, Saidi Kapiga 9,0 International Partnership for Microbicides, Clinical Affairs, Paarl, South Africa, 2 Desmond Tutu HIV Foundation, Masiphumelele Clinic, Cape Town, South Africa, 3 Maternal, Adolescent and Child Health, Edendale, South Africa, 4 MRC/UVRI Uganda Research Unit on AIDS, Masaka, Uganda, 5 Qhakaza Mbokodo Research Clinic, Ladysmith, South Africa, 6 Madibeng Centre for Research, Brits, South Africa, 7 Prevention for HIV/ AIDS Project, Pinetown, South Africa, 8 Ndlovu Care Group, Elandsdoorn, South Africa, 9 London School of Hygiene and Tropical Medicine, London, United Kingdom, 0 Mwanza Intervention Trials Unit, Mwanza, Tanzania, United Republic of Background: Multiple HIV prevention trials are being conducted in sub-saharan Africa as women remain vulnerable to HIV- infection. The dapivirine vaginal ring, currently being evaluated for safety and efficacy, shows potential as a topical microbicide to prevent HIV- infection in women. Methods: The Ring Study (IPM 027) is a Phase III, multi-centre, randomized, double-blind, placebo-controlled safety and efficacy trial of a dapivirine vaginal ring in healthy women. Initiated in March 202, HIV- sero-negative women, aged 8-45 years, are recruited at 7 research centres in South Africa and Uganda. Eligible participants must be sexually active, have normal serum chemistry and haematology profiles and a normal gynaecological examination. Women must not be pregnant, be on stable contraception, and not planning to become pregnant for the trial duration. Post-enrolment, participants attend monthly visits for HIV- counselling and testing, ring adherence counselling and are issued with an HIV risk-reduction care package and a new vaginal ring. STI tests are done 2-weekly and infections treated appropriately. Results: As of March 204, enrolment is ongoing with 483 women enrolled. The screen:enrolment ratio is 2:, the most common reason for screen failure being HIV-infection (up to 55%) and other sexually transmitted infections (up to 4%). The median age is 26 years with 57% < 25 years and 9% >35 years. At enrolment 64% reported secondary school education, and 6% tertiary education. 9% of the participants are unmarried, and 00% reported a primary partner in the last 3 months; 38% reported no condom use with the last sex act. The most commonly used contraceptive at baseline is long-acting injectable progestins (83%) followed by oral contraceptive pills (9%). Conclusions: It is imperative that HIV prevention methods be available to women to end the HIV epidemic. It is possible to recruit, screen and enrol eligible HIV-negative participants at risk for acquiring HIV into a Phase III microbicide vaginal ring trial. 46 HIV Research for Prevention 204 HIV R4P

417 Thursday, 30 October Posters 53: Vaginal Rings: Baseline Characteristics, Impact on Condoms and Flora P53.03 Baseline Characteristics of HIV- Negative Women Enrolled into a Clinical Trial of Dapivirine Vaginal Ring for HIV- Prevention P53.04 Effects of a Vaginal Ring Containing Maraviroc and or Dapivirine Worn for 28 Days on the Vaginal Microflora Thesla Palanee, Katie Schwartz 2, Elizabeth Brown 3, Vaneshree Govender 4, Nyaradzo Mgodi 5, Flavia Matovu Kiweewa 6, Gonasagrie Nair 7, Felix Mhlanga 5, Samantha Siva 4, Linda-Gail Bekker 8, Zakir Gaffoor 4, Nitesha Jeenarain 4, Sarita Naidoo 4, Francis Martinson 9, Jessica Phillip 4, Arendevi Pather 4, Bonus Makhanini 0, Lydia Soto-Torres, Jared Baeten 2, on behalf of the ASPIRE Study Team Wits Reproductive Health and HIV Institute, Johannesburg, South Africa, 2 FHI 360, North Carolina, NC, United States, 3 SCHARP - FHCRC, Seattle, WA, United States, 4 Medical Research Council, Durban, South Africa, 5 University of Zimbabwe-University of California San Francisco Collaborative Research Programme, Harare, Zimbabwe, 6 Makerere University-Johns Hopkins University Research Collaboration, Kampala, Uganda, 7 CAPRISA/University of Kwa Zulu Natal, Durban, South Africa, 8 Desmond Tutu HIV Foundation, Cape Town, South Africa, 9 UNC, Lilongwe, Malawi, 0 College of Medicine-John Hopkins University Research Project, Blantyre, Malawi, DAIDS/NIAID/NIH, Washington, DC, United States, 2 University of Washington, Seattle, WA, United States Background: Women s vulnerability to HIV- remains high in sub- Saharan Africa where transmission occurs mainly through heterosexual sex.antiretroviral prophylaxis, is a promising biomedical HIV- prevention strategy. Methods: ASPIRE - A Study to Prevent Infection with a Ring for Extended Use - is a phase III, randomized, double-blind, placebo-controlled trial testing the safety and effectiveness of the dapivirine vaginal ring for prevention of HIV- infection. Initiated in August 202, HIV- seronegative women between 8-45 years are being recruited from 5 trial sites in Malawi (MW), South Africa(SA), Uganda(UG), and Zimbabwe(ZIM). Eligible participants must be sexually active, have normal serum chemistry and hematology profiles, gynecologic examinations, not be pregnant nor planning to fall pregnant for the duration of participation. Post enrolment, participants attend monthly visits for HIV- counseling and testing, provision of a HIV- risk-reduction care package, adherence counseling, and provision of a new vaginal ring, to be worn continuously for the following month. Results: As of March 204, enrolment is ongoing with 2303 HIV- negative women enrolled in a screen:enrol ratio of 2.:. The median age is 26 years with 39%< 25 years and 4%>35 years. Overall, 00% reported having a primary partner in the 3 months prior to enrolment and 8% reported other partner. Eight percent of SA participants report being married, with 84% in ZIM, 82% in MW and 66% in UG. Forty percent report no condom use with the last sex act prior to enrolment. Chlamydia and gonorrhoeae prevalence is 4% and 4% respectively. Baseline contraceptive use reflects a wide method mix: 4% injectable depot medroxyprogesterone acetate, 9% contraceptive implants, 4% injectable Norethisterone enanthate, 3% intrauterine devices, and % oral contraceptive pills. Conclusions: African seronegative women at risk of HIV - infection can be successfully enrolled into a trial of dapvirine vaginal ring for HIV- prevention. Lorna Rabe, Leslie Meyn, Beatrice A. Chen 2, Lori Panther 3, Craig Hoesley 4, Sharon L. Hillier,2 Magee-Womens Research Institute, Pittsburgh, PA, United States, 2 University of Pittsburgh, Obstetrics, Gynecology, and Reproductive Sciences, Pittsburgh, PA, United States, 3 The Fenway Institute/Fenway Community Health, Boston, MA, United States, 4 University of Alabama at Birmingham, Birmingham, AL, United States Background: Vaginal rings with the capacity to release drugs over time may help with adherence to a microbicide for prevention of HIV. The objective of this study was to assess the impact of vaginal rings containing a microbicide on the microflora of the vagina and to assess the quantity of biofilm adherent to the rings. Methods: 48 Non-pregnant, HIV negative, sexually abstinent women aged 8-40 were enrolled in a double-blinded, 4 armed (2 per arm) randomized controlled trial. Women were recruited in Pittsburgh, PA; Boston, MA; and Birmingham, AL. Vaginal rings containing 25 mg dapivirine, 00 mg maraviroc, a combination of the two drugs, or placebo were worn for 28 days. The rings were removed and a subset of 6 was assessed for biofilm formation with electron microscopy. Vaginal swabs and smears were collected at baseline, 7, 28, and 52 days and cultured for aerobic and anaerobic bacteria and assessed for Nugent score. Modified Poisson regression and generalized estimating equations were used to assess the effect of ring use on the prevalence and concentration of vaginal microflora. Results: 47 women had cultures for at least 3 visits. The prevalence of pigmented anaerobic Gram negative rods increased significantly during the use of the placebo ring (RR 2.28, CI , P= 0.02) and dapivirine ring (RR.67, CI , P=0.04). Overall there was a decrease in prevalence of E. coli during ring use (RR 0.50, CI , P=0.04). However, no arm was statistically significant. There was no significant change of the Nugent score over the 52 days. The amount of biofilm ranged from scant to confluent but there was no significant difference in the quantity of biofilm stratified by the type of ring. Conclusions: Although there was an increase in prevalence of anaerobic pigmented Gram negative rods in the placebo and dapivirine users, there was no overall change in biofilm development or in Lactobacillus or microflora when assessed by Nugent score. 47

418 48 HIV Research for Prevention 204 HIV R4P

419 Author Index A Aaron, Erika Z....OA23.SY Abaasa, Andrew M.. OA02.04, P05.05, P09., P.04, P2.0, P36.0, P36.02, P40.2, P49. Abbai, Nathlee S....P4.09, P23.0, P26.0, P38.0, P38.05, P38.06, P39.04, P40.2, P49.0 Abbas, Ume...OA27.05 Abdool Karim, Quarraisha..OA9.06, P.02, P3.05, P23.06, P38.02, P40.2, P50.05 Abdool Karim, Salim S....OA06.05, OA2.0, OA2.02, OA30.03, PD05.02, P3.05, P34.08, P38.02, P40.03, P40.7, P40.9, P40.2 Abdullah, Fareed...RT04.03 Abdullahi, Hafiz...P3.08 Abebe, Kaleab...OA27.06 LB Abel, Kristina...P4.08 Abimiku, Alash le...p3.2, P30.05, P40.26 Abou, Max...PD02.02, PD02.05, P40.9 Aboud, Said...P26.08 Aboulker, Jean-Pierre...P26.0 Abrahams, Fatima...P34.03, P34.06 Abrahams, Melissa-Rose...P25.09 Abullahi, Bala...P3.08 Abuogi, Lisa...P45.08 Acharya, Priyamvada...OA0.06, P0., P5.2 Achilles, Sharon L....OA20.05 LB Achiro, Lilian...P30.04 Ackerman, Margaret E....OA20.03, P03.04 LB, SY06.02 Ackland, Jim...PD03.04 LB Adam, Barry...P48.03, P48.05 Adam, Lucille...P27.02 Adams, Andrew E....P39.0 Adams, Cameron...P34.3 LB Adams, Elizabeth...P26.02 Addo, Marylyn M....P24.8 Adebajo, Sylvia...P09.09, P09.2, P49.04 Adedokun, Olanrewaju...P43.0 Aderem, Alan...OA6.06 Adetokunboh, Olatunji O.... P3.0, P45.0, P45.02 Adhikary, Rajatashuvra...P28.0 Aerts, Joeri L....P27.0 Agot, Kawango...OA5.05, P46.0, P46.02 Aguiar, Renato...P5.03 Aguilar-Jiménez, Wbeimar... P04.02, P2.5, P37.0 Ahani, Bahar...P25. Ahlers, Jeff...P4.30 LB Ahmed, Hasan...OA08.04, P26.4 Ahmed, Khatija... OA5.05, P2.03, P46.0, P46.02, P49.03, P50.0 Ahmed, N...P05.08 Ahmed, Nurelign...P3.0 Ahmed, Nuri...OA28.04, P49.08 Ahmed, Nurilign...P05.03, P09.0, P3.02, P42.0 Ahmed, Tina...P26.05 Ahonsi, Babatunde...P09.2, P49.04 Ajbani, Seema...P4.0 Akanni, Olayide...P07.0 Akapirat, Siriwat...OA.05, P26.3 Ake, Julie...P23.02, P26.02 Akello, Carolyne A....OA5.02, P05.0, P42.02, P42.05 Akolo, Maureen...P9.0 Alagbe, Sikeade...P3.3 Alaka, Oluwatosin B... P07.0, P02.08 Alam, Munir...P0.2 Alam, Muntasir...P34.05 Alam, S. M....OA2.06 LB Alcamí, José...OA22.05, P35.04, P4.02, P44.04, P44.0 Alcantara, Sheilajen...P25.0 Aldunate, Muriel...P40.0, P40.29 Alexander, David L....P0.0 Alexander, Mallika...P48.0 Alexandre, Kabamba B.... P33.0, P33.03 Algate, Paul...OA2.02 Alhassan, Emmanuel...OA28.05, PD0.03 Alicea, Candido...OA6.05, OA24.04, OA29.06 Aling, Emanuel...P05.05, P09. Alleman, Patty...P4.0 Allen, Mary...OA05.02, OA0.03, OA.04, P42.0 Allen, Shannon...P25.02 Allen, Susan...RT04.05, OA.0, OA4.0, OA20.06, OA2.0, OA2.03, OA2.04, OA28.04, P0.0, P05.03, P05.08, P08.0, P09.0, P09.05, P3.02, P3.0, P3.4, P23.6, P24.2, P36.06, P39. LB, P40.22, P42.0, P48.02, P49.08, P52.0 Allen, Todd M....SY02.04, OA2.05, P2.03, P24.9 Almeida, Rafael...P4.04 Almond, Neil...P5.0 Alonso, Esther...P33.05, P5.04 Alonso, Maria... P4.7, P4.8 Alonzi, Dominic...OA8.02 Alt, Carsten...P5.04 Alter, Galit...SY06.02, SY.04, OA2.05, OA24.02, OA24.03, OA25.02, PD05.03, P03.04 LB, P6.07, P34.2 LB, P4.23 Altfeld, Marcus...OA04.0, OA4.0, OA2.03, OA2.05, P2.03, P2.6, P2.7 Alvarez, Susana...P5.03, P5.04 Alvarez-Fernández, Carmen..OA8.03 Alving, Carl R....P0.08 Aman, Rashid...P22.05, P25.04 Amara, Rama...OA08.05, OA29.05 LB, PD03.02 Amarasena, Thakshila H....P25.0 Ambada, Georgia...P2., P2.4, P4.22, P52.02 Ambrozak, David R OA06.04 Amenichi-Enahoro, Susan...P52.04 Amero, Molly...OA2.05, P2.03 Amico, K. Rivet...P9.02 Ampofo, Stephen...P8.0, P43.08 Amuamuziam, Augustina O....P02.08 Amuamuziam, Obiajulu A P07.0 An, Dong Sung...OA05.0 Anahtar, Melis...OA20.04, P40.08 Ananworanich, Jintanat...OA05.04 Ancuta, Petronela... OA04.06 Andayi, Freda...P45.06 Andersen-Nissen, Erica...OA.06 LB Anderson, Deborah J....OA24.02, P40.02, P40.29, P4.9 Anderson, Sarah...P23.6 Anderson, Sharon...P40.30, P44. Andersson, Soren...P26.6 Andrasik, Michele P.. OA9.0, P26.09 Andreeva, Vladanka...P09.04 Andrews, Charla...P44.09 Anenih, James...PD0.03 Angin, Mathieu...P24.8 Angus, Brian...P26. Aniagyei, Stella E....OA26.04 Ansari, Aftab A....OA7.02 Anthony, Colin...P39.05 Anton, Peter...P23.03 Antony, Joseph M....OA6.03 Anude, Chuka... P07.02, P26.09 Anzala, Omu... OA.0, OA7.03, PD03.04 LB, P0.0, P24.09, P26.04, P26.07, P40.23, PL03.03 Aoko, Appolonia...P23.02 Apaka, Cosmas...P49.0 Apidi, Winnie...P37.02 Appel, Cara...OA7.05 Appiagey, Ashey...P05.08, P09.0 Arakelyan, Anush...OA30.06 LB Aravantinou, Meropi...OA03.05, OA0.02 Archary, Derseree... P40.03, P40.09, P40.7 Archer, David F....OA3.03, P23.5 Archer, Eva J....OA06.0 Arcia, Eliuth D....P04.02 Ardón, Elvia...P22.02 Arien, Kevin...P35.0, P39.03, P40.5, P44.05 Arnold, Kelly...OA0.04, OA2.05, P40.9 Arroyo, Miguel A.... OA08.04 Arthos, James...OA7.02, OA7.03, P34.02, P39.06 Asbach, Benedikt...P0.07 Ashfield, Rebecca OA05.05 Asiimwe, Stephen...OA28.0, OA28.02, P05.06, P23.07 Asiki, Gershim...P09., P.04, P2.0, P36.0, P36.02, P49.02 Asin, Susana...P44. Asin-Milan, Odalis... OA04.06 Assob, Jule Clement Assob N...P2. Augustyns, Koen...P35.0 Auñón, David...P4.02, P44.0 Avenant, Chanel...OA20.02 LB, P05.09 LB, P33. LB Avila-Rios, Santiago...P22.02 Awuor, Nicollate...P36.08 Axthelm, Michael K....SY.03 Ayehunie, Seyoum...P40.02 Ayles, Helen...OA28.03 Ayub, Snaidah O....P46.05 Ayuo, Paul...P49.0 B Baalwa, Joshua...OA.0 Back, David...OA27.0, OA27.06 LB Baden, Lindsey..OA06.02 LB, P26.03 Badial-Hernandez, Florentino..P06.0 Baeten, Jared... OA27.02, OA28.0, OA28.02,PL0.02, PD0.0, PD02.0, P02.0, P03.0, P09.7 LB, P20.03, P24.07, P46.06, P52.06, P53.03 Baggaley, Rachel...P09.04 Bahati, Prince N....OA9.02 Bahemuka, Ubaldo...P09., P.04, P2.0, P36.0, P36.02 Bailer, Robert...OA6.0 Bailey, Adam...OA4.06 Bailey-Kellogg, Chris... SY06.02, P03.04 LB Bailor, Bob...P4.30 LB Bakanda, Celestine...P52.08 Bakari, Muhammad... P26.08, P4.2 Baker, A Cornelius... P02.07, P43.04 Baker, Jonathan R....P23.8 Bala, Manju...P2.0 Bala, Veenu...P44.0 Balán, Iván...P42. Balazs, Alejandro B....OA05.0 Bale, Shridhar...OA0.02 Balestre, Éric...P52.05 Baleux, Françoise...P44.05 Balkus, Jen...P49.05 Balkus, Jennifer E....P0.02, P05.0, P36.03, P36.04, P45.05 Ball, Cameron...OA26.05 Ball, Terry Blake...OA08.03, OA0.04, OA7.04, PD02.0, PD02.02, PD02.03, PD02.05, P03.02, P24.04, P24.05, P24.3, P37.02, P37.08, P40.3, P4.7, P4.8 Balogun, Tolulope...P45.02 Baltimore, David...OA05.0 Banchereau, Jacques...P24.0 Banda, Helman...P09.0 Bandivdekar, Atmaram...P4.0 Bangsberg, David R....OA23.02, OA23.03, P37.03, P43. Banki, Zoltan...P4.2 Bansal, Anju... P24.7, P26.7 Bansal, Manish...OA0.04, P6.06, P34.0, P4.26 Barbizan, Thais...P5.03 Barcena, Luis...OA04.04, P25.02 Barin, Burc... PD03.04 LB, P26.07, P26.2 Barin, Francis...P39.02 Barinaga, Guillermo...P27.03 Barnabas, Ruanne...P09.7 LB Barnabas, Shaun L.... OA7.0, P3.03, P40.4 Barnable, Patrick...P4.02 AUTHOR INDEX 49

420 Author Index AUTHOR INDEX Barnett, Susan W....OA06.03, OA08.06 LB, OA25.0, P03.04 LB, P0.09 Barnighausen, Till...SY03.03 Baron, Deborah...OA9.05, P23.08 Barouch, Dan... OA04.0, OA06.03 Barragan, Fatima...P40.28 Barré-Sinoussi, Françoise... P2.0, P40.20 Barrows, Brittani...OA23.0 Barry, Christina... P4.7, P4.8 Bashford-Rogers, Rachael...OA06.0 Bashir, Tahir...OA8.0 Bass, Emily... PD06.02 Basu, Debby...P39. LB Basu, Rahul...OA05.03 Bates, Adam...P25. Batorsky, Rebecca...OA2.05, P24.9 Baty, Daniel...P39.02 Bauer, Asli...P26.08 Bauman, Laurie J....P5.02, P23.7 Baumgartner, Joy N....P3.5 Bautista-Arredondo, Sergio...P06.0 Baxa, Ulrich...OA0.0 Baxter, Cheryl...P38.02 Bayingana, Roger...OA28.04, P05.03, P26.03 Bazner, Sue...OA2.05 Beamer, May A....OA20.05 LB Beaudoin, Leon...P8.05 Becker, Ericka...P0.06 Bedard, Hunter...OA2.05 Bedoya, Luis-Miguel...P4.02 Beigi, Richard...P45.05, P45.07, P50.04 Bekker, Linda-Gail...OA09.05, OA0.03, OA.04, OA7.0, P3.03, P26.03, P26.2, P42.0, P50.0, P53.02, P53.03 Beksinska, Mags...OA23.04, OA26.0, P30.06 Bellier, Bertrand...P27.05 Beltran, Manuela...P4.02 Bender, Bonnie...PD0.05 Benki-Nugent, Sarah...P22.0 Bennett, Jane...P43.4 Bennett, Kara...OA23.02 Bennett, Kathleen...P34.09 Bennie, Thola...OA7.0, P3.03 Benny, Thola...OA5.04 Benyeogor, Ifeyinwa...P40.0 Berberich, Matthew...P24.20 LB Bergin, Philip... PD03.04 LB, P26.04, P40.22, P40.23 Beri, Archana...P36.05 Berkhout, Ben...P39.03, P5.0 Berman, Phillip W P0.0 Bernard, Nicole... OA04.06, OA4.02 Berry, Neil...P5.0 Berry, Susan...P3.0, P3.02 Bershteyn, Anna...P20.04 LB, P43.5 LB Bertniz, Netanya...P39.06 Bertram, Melanie P47.06 Berzi, Angela... PD02.04 Betts, Michael...OA24.0 Bewley, Carole...P34.3 LB Beyrer, Chris...PL02.0 Bhasin, Manoj K....P24.8 Bhat, Hans R....P33.08 Bhatia, Gaurav P5.04 Bhatta, Rabi S....P44.0 Bhattacharya, Jayanta...OA0.04, P6.06, P34.0, P4.26 Bhiman, Jinal N....OA06.05, OA2.0, PD05.02 Bhosale, Ashok V....P8.07 Biasin, Mara... OA08.02, PD02.04 Biberfeld, Gunnel...OA.02, P26.08, P26.6 Bibollet, Christelle...P09.4 LB Biedma, Marina...P6.05 Bielawny, Thomas...OA08.03, P4.7, P4.8 Biello, Katie B....P08.03 Bigert, Matthew...P4.05 Biggio, Joseph...P45.07, P50.04 Billings, Erik...OA25.0, P03.04 LB Binagwaho, Agnes...RT03.0 Binello, Nicolo...OA06.03, OA25.0, P03.04 LB Binley, James M.... OA06.04 Biron, Julie P09.4 LB Birse, Kenzie...OA0.04, PD02.0, P40.04, P40.9 Birx, Deborah L....PL04.0 Bitan, Gal...P44.07 Bitangumutwenzi, Patrick...P30.0 Bizimana, Jean... PD03.04 LB, P3.02 Bjorkman, Pamela J.... SY05.0, P34.09, P39.02 Blackard, Jason T....P38.02 Blackburn, Matthew...OA06.03, OA25.0, P03.04 LB Blakney, Anna...P8.02 Blanchard, James...OA0.02 Blanchard, Kelly...P09.0 Blanco, Juliá...P6.0, P6.04 Blanda, Wendy... OA26.03, P4.03, P4.06, P4.07, P4.08 Blaschke, Terrence F....SY04.04 Blass, Eryn...OA04.0 Bockh, Emily. P02.07, P2.04, P2.05, P48.06 Boczar, Ashlee D....P5.9, P5.23 Bodwell, Jack E....P5.24 Boesch, Austin W....OA20.03 Boffito, Marta...OA27.0 Boige-Faure, Sylvaine...P09.4 LB Boily, Marie-Claude... SY04.03, P20.0 Bokhart, Mark...OA26.06 LB Boliar, Saikat...OA0.04, P6.06, P34.0 Bolton, Diane L....SY.04 Bonaventura, Clotet...P24. Bonduelle, Olivia...P2.02, P2.08 Bonnaffé, David...P44.05 Booley, Mumtaz PD03.04 LB Boonchawalit, Samatchaya...P35.03 Borggren, Marie...P4.0 Borrow, Persephone...OA.0 Borthwick, Nicola J....P26.05 Bose, Meera...OA08.04, OA2.06 LB, P25., P39.07 Bossert, Adam...P0.03 Bossi, Giovanna...OA05.05 Bouakane, Amel...P26.0 Bouchemal, Kawthar...P44.05 Boum, Yap...P43. Bourguignon, Patricia...P26.07 Bouvin-Pley, Melanie...P39.02 Bowman, Brittany...OA20.04, OA24.03 Bowman, Christine...OA07.03 Boyd, Peter... OA26.03, P4.03, P4.04, P4.06, P4.07, P4.08, P5.4 Braakman, Ineke...OA8.02 Brache, Vivian...OA3.03 Bracke, Lotte...P39.03 Brady, Martha M....P09.6 LB Brady, Michael...OA07.03 Bragg, Vivian...P42.05 Braibant, Martine...OA23.0, P39.02 Brander, Christian...OA4.02, OA6.05, OA29.06, P24.03, P27.0 Branson, Bernard...P06.03 Bråve, Andreas...OA.02 Breaker, Brett...P4.05 Brener, Jacqui...P24.9 Breton, Gaelle...P4.30 LB Brezar, Vedran...P24.0 Bright, Danielle...P5.8 Brill, Ilene...OA20.06 Brimer, Andrew...OA26.03, P4.03, P4.04, P4.05, P4.06, P4.07, P4.08 Briney, Bryan...OA0.03, OA2.02 Brochard, Patricia...OA03.04 Brockman, Mark...OA09.03, P09.05, P24.2, P33.06, P37.03, P48.02 Brod, Florian...OA25.05 Broder, Gail B....OA9.0, P07.02, P26.09 Broderick, Kate...P0.0, P26.5 Broliden, Kristina... OA7.04, PD02.03 Brostrom, Martina...OA07.02, P D04.04, P09.04 Brouckaert, Peter...OA24.03 Brown, Alison E....SY03.04 Brown, Ben... OA09.05, PD04.05 Brown, Elizabeth...RT05.04, P36.03, P42.03, P53.03 Brown, Eric...P03.04 LB Brown, J. Kendall...P44.09 Brown, Sheldon T....P06.02 Brown, Stephen J....OA05.03 Brown III, William... PD04.03 Brumme, Chanson...OA4.04 LB, P37.03 Brumme, Zabrina...OA4.04 LB, P37.03, P39.0 Brummer, Zabrin...P33.06 Brumskine, William...P50.0 Bruun, Tim-Henrik...P4.27 Buchbinder, Susan P....OA.03, PD06.03, P26.4 Buchholz, Caitlin A....P5.23, P8.03 Buckheit, Karen W.... P5.9, P5.23, P8.03, P33.0, P44.02 Buckheit, Robert...PD04.02, P5., P5.2, P5.9, P5.23, P8.03, P33.0, P44.02 Buhler, Shayna...P48.03, P48.05 Buisson, Sandrine...OA05.05 Bukenya, Regina...P23.09 Bukusi, Elizabeth A....RT05.05, OA02.0, OA28.0, OA28.02, P03.0, P24.07, P30.04, P36.08, P45.08, P48.04 Bulya, Nulu... OA28.0, OA28.02 Bunge, Katherine.. OA0.0, OA3.0, P05.0, P45.07, P49.07 Burgener, Adam OA0.04, PD02.0, PD02.05, P2.06, P40.04, P40.9 Burger, David M....P22.03 Burgers, Wendy A....P25.09 Burgess, Jacqueline S....P43.02 Burns, Julien...P47.02 Burton, Dennis R....OA0.03, OA2.02, OA25.04 Burton, Samantha...OA08.05, PD03.02 Burwitz, Benjamin J....SY.03 Busch, Michael OA.0 Byakwaga, Helen...P37.03 Byrne, Elizabeth H....OA20.04 C Cabello, Robinson...P52.03 Caccuri, Francesca...OA04.04, OA06.03, OA25.0, P03.04 LB Caceres, Carlos F....P08.02, P43.03 Cahn, Pedro...PD05.04, P24.5 Caillon, Patrick...P09.4 LB Cain, Brian...OA4.06 Cairns, Gus...P43.04 Calamante, Gabriela...P4.07 Calandra, Thierry...P0.07 Cale, Evan M.... OA06.04 Callahan, Marianne...P36.05 Cambau, Sébastien...P09.4 LB Cameron, Mark...OA04.05 LB, OA25.0, P03.04 LB, P4.30 LB Campbell, Russell D....P07.05 Cannou, Claude...P40.20 Cao, Shijie...P5.8 Capa, Laura...P4.02 Capina, Rupert...OA08.03, P37.08, P4.7, P4.8 Cara, Andrea...SY09.04 Carbajal, Candy...P22.02 Carballo-Dieguez, Alex...PD04.03, P3.7 LB, P42. Carbonetti, Sara...OA0.05, OA2.03 Cardozo, Timothy...P26.8 Carfi, Andrea...P0.09 Carias, Ann M....P40.05 Carlson, Jonathan...OA08.04, OA4.04 LB, OA2.0, P24.7, P26.7, P HIV Research for Prevention 204 HIV R4P

421 Author Index Carrasco, Esther OA8.03 Carreras, Esther...OA8.03 Carrico, Chris... OA08.04, OA2.04 Carrillo, Jorge...OA29.06, P6.0, P6.04 Carrington, Mary...OA29.04, P37.03 Carstens, Hannelie P53.02 Carter, Allison...P53.02 Carter, Darrick...P27.03 Carulei, Olivia...P4.24 Caruz, Antonio...OA08.02, P37.0 Cascioli, Roberto...P09.03 Caskey, Marina...P4.30 LB Castellano, Laura...P44.07 Castello, Stéphane...P09.4 LB Cavarelli, Mariangela.SY08.03, P03.03 LB Cefola, Raymond...P23.03 Cekwane, Themba...P40.09 Cele, Naureen...P05.07 Celum, Connie...RT03.03, OA27.02, OA28.0, OA28.02, PD02.0, P03.0, P05.0, P09.7 LB, P20.03, P24.07, P46.06, P52.06 Center, Robert...P0.4 LB Centlivre, Mireille...P2.02 Cepeda, María Victoria...P0.07 Ceulemans, Ann...P24.4 Chackerian, Bryce...PD03.05 LB Chaikummao, Supaporn... P09.3, P30.02, P36. Chakrabarti, Bimal K....OA0.04, P6.06, P34.0, P4.26 Chamberland, Annie... OA04.06 Chames, Patrick...P39.02 Chan, Brian T....P43. Chan, Jacqueline K....OA6.03 Chandhiok, Nomita...P36.05 Chandipwisa, Adlight...P4.0 Chandra, Neelima...OA0.05, P 40.30, P44. Chang, Judy...P2.6 Chan-Hui, Po-Ying...OA2.02 Chapman, Ros...P4.09 Chapon, Catherine...P27.02 Chappell, Catherine...OA0.0, P49.07 Chariyalertsak, Chonlisa...P09.02 Chariyalertsak, Suwat... P09.02, P44.09 Charles, Kelly P5.26 LB Charls, Patrick...P32.0 Charuwat, Chutima...P09.02 Chatani, Manju...P43.05 Chataway, Joanna...P9.03 Chaudhary, Nakul K....P6.06 Chaudhary, Omkar...P2.0 Chauhan, Sanjay...P36.05 Chavez Baray, Silvia...P3.7 LB Chawda, Mira M....P26.06 Cheeseman, Hannah M....P40.06 Chege, Gerald...P25.06 Chemnasiri, Tareerat... P30.02, P36. Chen, Beatrice A....OA3.03, OA3.05 LB, P53.04 Chen, Joseph...P40.28 Chen, Lei...P0.02, P0. Chen, Li-Qing...OA6.02 Chen, Pai-Lien...P25.0 Chen, Xuejun...P0.03 Chen, Xuemin...P26.9 Chen, Yanli...P6.02 Chen, Zhiwei...P4.4, P4.29 Cheng, Cheng...OA0.06, P0.03 Cheng, Helen...OA5.04, P42.04 Cheng, Lin...P4.29 Chenine, Agnès-Laurence... SY.04, OA08.0, OA2.06 LB, OA23.0, P39.07 Cherutich, Peter...RT03.02 Chetty, Paramesh... PD03.04 LB, P26.04 Chetty, Shivan...P4.09 Chhonker, Yashpal S....P44.0 Chianese, Christopher...OA09.06, P9.02 Chidiac, Christian...P09.4 LB Chidrawar, Shweta...P48.0 Chihota, Emilder...P23. Chilende, Clever...P43.06 Chinga, Jonah M....OA9.02 Chinnawirotpisan, Piyawan.... P25.08 Chinyenze, Kundai...PD03.04 LB Chiodi, Francesca...OA7.0, P3.03 Chipato, Tsungai...P4.0, P25.0 Chirenje, Z. Mike...OA0.06 LB, OA20.05 LB Chirenje, Zvavahera M....OA5.03, P05.0, P4.0, P23., P36.03, P36.04, P42.03, P49.05, P49.06 Chisembele, Maureen...OA02.04, P40.2 Chissumba, Raquel...P26.6 Chitwarakorn, Anupong... OA07.05, P09.3, P3., P30.02, P36., P49.2 Chohan, Bhavna H....P22.0 Choi, Min Joung...P27.06 Choji, Grace...P3.2, P30.05 Chomba, Elwyn...OA20.06, P52.0 Chomchey, Nitaya...OA05.04 Chonco, Sthe...P30.06 Chonwattana, Wannee...P49.2 Chou, Shih-Feng...OA26.05 Choudhary, Ipsita...P6.06 Chrisman, Cara J....RT05.0 Christopher, Miller...OA25.0 Chuchuen, Oranat...OA22.04 Chung, Amy...OA24.02, P4.23 Chung, Amy W....PD05.03, P03.04 LB Chuong, Dinh...OA03.06 LB Churchill, Melissa J....P35.02 Churchyard, G....OA06.02 LB Churchyard, Gavin...OA0.03, OA.04, P42.0 Cianci, Gianguido C....P25.02 Cicala, Claudia...OA7.02, P34.02, P39.06 Cimbro, Raffaello...OA7.02, P34.02 Claiborne, Dan OA.0 Claiborne, Daniel...OA4.0, OA2.0, OA2.03, OA2.04 Clark, Justin...OA03.03, P5.6 Clark, Lorna...PD03.04 LB Clark, Meredith...OA03.03, P4.02, P5.6 Clarke, Amanda...OA07.03 Clarke, William...OA07.06 LB Clemente, Maria Isabel...P5.03 Clerici, Mario... OA08.02, PD02.04 Climent, Núria...OA8.03, P24.03, P4.03 Clotet, Bonaventura...OA29.06, P6.0, P6.04 Cobarrubias, Kyle...P37.03 Cobb, Karen...P26.06 Cochrane, Alan...OA20.0 Cochrane, Gavin...P9.03 Coetzee, David...OA04.03, P40.4 Coetzee, Jenny...P29.0 Cohen, Craig R....PD02.0, P36.08, P45.08 Cohen, Myron S....OA22.0 Cohen, Sarah S....P38.03 Coic, Yves-Marie...P44.05 Cokley, Cheryl...P02.04 Cole, Alexander M....P44.08 Cole, Kelly S....PD03.02 Colebunders, Robert...P24.4 Colizzi, Vitorro...P4.22 Collins, Clare...PD0.04, P02.02 Collman, Ronald G....PD03.02 Colvin, Christopher J....OA02.05, P46.03 Combadiere, Behazine... P2.02, P2.08 Come, Jotamo...P29.05 LB Conde-Glez, Carlos J....P06.0 Cone, Richard... P40.0, P40.6, P40.29 Connell, Bridgette J....P44.05 Connole, Michelle...P2.09 Connors, Mark...OA6.0, P5.25, P24.06 Constantine, Brian...P33.0 Cools, Piet...P40.5 Cope, Alethea...OA08.06 LB Cope, Alethea V....P26.06 Coppens, Sandra...P39.03 Corbelli, Giulio Maria...P09.03 Corbett, Jennie...P9.03 Corcoran, Padraig...P52.04 Corey, Larry...OA06.02 LB, OA0.03, OA.06 LB, PD06.0 Corleis, Bjorn...P2.03 Corneli, Amy... OA5.05, P.0, P46.0, P46.02 Cornelissen, Marion...P39.03 Correia-Pinto, Jorge.. P4.7, P4.8 Cortez, Valerie...P26.9 Cosgrove Sweeney, Yvonne...P5.20 Costanzo, Margaret...OA2.06 LB Cotton, Laura...OA09.03 Cottrell, Mackenzie L....OA22.06 LB Coudeyras, Colette P09.4 LB Courtney, Colleen..P6.03, P34. LB Coutinho, Alex G....PL04.02 Coutinho, Helen...P26.04 Cox, Josephine PD03.04 LB, P24.09, P26.03, P26.04, P26.07 Cranston, Ross D.... SY08.04, OA27.06 LB, PD04.03 P23.03, P42. Creasy, George...P5.27 LB Crespo, Manel...OA29.06 Crispin, Max...SY05.02 Crook, Angela...OA02.04, P40.2 Crooks, Ema T.... OA06.04 Crotty, Shane...OA.0 Crucitti, Tania...P40.5 Cubas, Rafael...OA29.0 Cumbane, Victória...P26.6 Cummings, Beverley...P29.05 LB Cunha, Rodrigo...P5.03 Cunha-Neto, Edecio...P4.04 Cunningham, Pamela...OA05.03 Cunningham, Tina D....OA0.05 Cupo, Albert OA25.04 Curlin, Marcel...P25.08, P49.2 Curran, Kathryn...P09.04, P46.06 Curran, Kelly...P29.05 LB Curriu, Marta...P6.04 Cuthbertson, Jack...P0.4 LB Cu-Uvin, Susan...P40. Cuzin, Lise...P26.0 Czarnecki, Chris... P4.7, P4.8 Czyzewska-Khan, Justyna...P26.06 D Da Silva, Luis...P5.03 Dabee, Smritee...OA7.0, P3.03, P40.07 Dabis, François...P52.05 Dadem, Nancin Y....PD0.03 Dahlke, Christine...P24.8 Dai, James...OA0.06 LB, P42.05, P45.07, P49.06, P50.04 Daitiri, Ruth...P30.05 Dally, Len...PD03.04 LB, P26.03 Dang, Danielle...P05.08, P42.0 D Angelo, Lawrence J....P06.04 Daniuk, Christina...OA08.03, P4.7, P4.8 Dantas, Ezequiel...P24.02 Darko, Sam...P0.2 Das, Atze...P5.0 Das, Supratik...OA0.04 Das Neves, José...P5.22 Dash, Pradyot...P24.6 Datiri, Ruth...P3.2 Datong, Pam...P3.2, P30.05 Davis, Benjamin G....OA25.05 Davis, Isaiah...OA29.04, P24.0 Davitte, Jonathan...P09.05, P48.02 Dawood, Hassan Y....OA0.05 Dawson, Liza...P45.03 Dayton, Robyn...OA02.0 De Gioia, Luca...OA08.02 De la Mata, Francisco J.... P5.0, P33.04, P33.05 AUTHOR INDEX 42

422 Author Index De Luca, Mariacristina PD02.04 Diaz, Laura...P5.03, P5.04 Duba, Nosipho... OA02.02, PD0.02 Emerole, Karl...P52.07 AUTHOR INDEX De Oliveira, Tulio...SY03.03 De Rosa, Stephen...OA04.05 LB, OA6.06, OA7.06, PD03.0 LB, P24.08 De Rose, Robert...P25.0 De Souza, Mark... P25.08, OA05.04, OA.05 De Truchis, Claire...P2.0, P40.20 De Val, Natalia...OA0.02, P0.04 De Villiers, Marthie...OA09.02, P9.05, P23.04 Deal, Cailin...OA05.0 Debo, Brian M....OA0.0, P49.07 Debora, Bossemeyer...P29.05 LB DeCamp, Allan C.... OA08.04 Decoville, Thomas...P6.05, P6.08 Dedes, Nikos...P43.04 Deffur, Armin...P4.24 Degli Esposti, Michele...P09.03 Del Medico Zajac, Maria Paula. P4.07 Delaloye, Julie...P0.07 Delany-Moretlwe, Sinead... SY04.02, OA9.05, OA27.03, P23.08, P38.03, P40.2, P40.5, P50.0 Delinotte, Anne-Cécile...P09.4 LB Delisle, Josie...P40.0 Dellar, Rachael C....P.02 Dellon, Evan S....OA22.06 LB Delpech, Valerie C....SY03.04 Dennis, Kristine...OA2.0 Denny, Thomas...OA.0 Derby, Nina...OA03.05 Derdeyn, Cynthia.. OA08.05, PD03.02 Dereuddre-Bosquet, Nathalie... OA03.04, P44.05 Derking, Ronald...OA8.02 DeRosa, Steve...OA.06 LB Derrick, Tiffany... OA26.03, P4.05, P4.06, P8.0 Des Jardins, Delphine...OA08.06 LB Desai, Monica...OA07.03 Desaint, Corinne...P26.0 Deshpande, Sucheta..P09.08, P48.0 Deshpande, Suprit...P4.26 Desjardins, Delphine...OA03.04, P44.05 Desmond, Nicola...OA07.02 DeSouza, Mark...OA08.04, P26.02 DeVico, Anthony L....OA25.03, OA30.02, SY09.02 Devlin, Brid...OA3.0, OA22.02, OA26.03, P4.03, P4.04, P4.05, P4.06, P4.07, P4.08, P8.0, P43.08, P44.03 Deymier, Martin J....OA2.04 Dezzutti, Charlene...OA3.0, OA3.03, OA3.05 LB, OA22.02, OA22.03, OA27.06 LB, OA30.04, P5.05, P5.9, P5.23, P8.03, P44.03 Dhitavat, Jittima...P44.09 Dhlakama, Patricia M....P23. Di Pilato, Mauro...P4.05 Diallo, Dazon D....P43.07 Diaz, Gabriela...OA7.06 DiazGranados, Carlos...OA.06 LB Diefenbach, Thomas...OA24.03 Dietrich, Janan...OA09.03, P3.03, P29.0 Dil, Nyla...P40.3 Dilernia, Dario...OA2.0 Dimitrov, Dimiter S....OA6.02 Dimitrov, Dobromir...PD06.0, P20.0 Ding, Ming...P40.27 Dingens, Adam S....P26.9 Dionne, Kendall...OA25.02 Dippenaar, Nicole...P23.04 Diskin, Ron...SY05.0 Dispinseri, Stefania... OA08.06 LB, P03.03 LB Dissen, Elisabeth...P09.05, P48.02 DiStefano, William...P0.03 LB Dittmer, Ulf...SY.04 Djebali, Sarah...OA24.06 LB Dlamini, Cynthia...OA9.05 Dlamini, Sarah..... OA02.03, P40.03, P40.7, P50.03, P50.05, P50.06 LB Dlungwana, Thuleleni...P38.03 Dobard, Charles...OA03.06 LB Dodge, Brian...P3.7 LB Dodzo, Munyaradzi Kenneth...P05.02 Doglioni, Claudio...P44.0 Doherty, Kathleen...OA20.04, P40.08 Dolling, David...OA07.03 Dominguez, Karen...OA09.05 Dommaraju, Kalpana...P25. Donaldson, Emily D....PD06.02, P47.0, P47.02, P47.03, P47.07 Donaldson, Mitzi. OA25.0, P03.04 LB Doncel, Gustavo F....OA0.05, OA3.03, OA3.04, P23.5, P4.02, P25.0, P33.03, P40.30, P44. Dong, Krista...OA20.04, OA29.02, P34.2 LB, P40.08 Donnell, Deborah...OA28.0, OA28.02, P03.0, P20.03, P24.07 Dorfman, Jeffrey R.... P25.09, P34.03, P34.06 Doria-Rose, Nicole A....OA06.04, OA2.0, PD05.02 Dorrell, Lucy...OA05.05, OA29.06, P26.05, P26. Doster, Melvin N....OA04.04, OA06.03, OA25.0, P03.04 LB Dott, Clare... P43.02, P47.04 Douek, Daniel...OA06.0, OA4.0, OA6.04, P0.2 Douglass, Nicola...P4.24 Douglass, Niki...P4.09 Douville, Renee...P03.02 Dowell, Karen...P03.04 LB Drannik, Anna G....P40.26 Dreyer, Anita M....OA0.05 Druz, Alex...P5.2, P5.25 Druz, Aliaksandr...OA0.0, OA0.06, P5.28 LB Du, Victor Y....P24.7 Du Toit, Andrea...P05.09 LB, P33. LB Duan, Song...P39.0 Duby, Zoe...OA02.05, OA02.06 LB, P46.03 Ducloy, Camille...P6.08 Duerr, Ann...OA7.06, P52.03 Duerr, Ralf...P34. LB Duette, Gabriel...P24.02 Duffill, Kathy...OA27.06 LB Duffy, Ryan...OA2.06 LB Dugast, Anne Sophie...OA2.05, P34.2 LB Dulai, Joshun...P48.03, P48.05 Dumont-Blais, Alexandre... P48.03, P48.05 Duncan, Renee C....P35.02 Dunn, David...OA07.03 Dunphy, Laura...P40.9 Durgiah, Raveshni...OA30.03 Durier, Christine...P26.0 Duriez, Marion...P2.0, P40.20 Durueke, Florita...P02.08, P02.09, P07.0 Dwek, Raymond...OA8.02 E Eakle, Robyn... P3.04, P46.04, P48.07 Eamsila, Chirapa...P26.3 Earl, Patricia...OA.02, P26.02 Easterhoff, David...OA2.06 LB Eckhoff, Philip...P20.04 LB, P43.5 LB Ecuru, Julius...P07.03 Edlefsen, Paul T.... OA08.04 Edoni, Elizabeth...P45.04 Edward, Vinodh...OA.0 Edwards, Gregory...P0.03 LB Egami, Yoshika...P34.05 Egan, Michael...OA05.02, P26.07 Egerer, Lisa...P4.2 Egessa, Aggrey...P52.08 Ehrilich, Nathan...OA3.0 Eisinger, Robert W....PL04.0 Ekeruche-Makinde, Julia N....P5.26 LB El Costa, Hicham...P2.0, P40.20 Elamin, Ayssar...P26.06 Eldridge, John...OA05.02, P26.07 El-Far, Mohamed... OA04.06 Elharrar, Vanessa...P0.02 Elion, Richard...P52.04 Elizaga, Marnie...OA05.02, OA.03 Eller, Leigh Anne...OA2.06 LB, P25., P39.07 Eller, Michael...OA2.06 LB, SY.04 Ellet, Anne...P35.02 Ellington, Sascha...P03.05 LB Elliot, Debra H....OA06.05 Elliot, Tim...P26.05 El-Omar, Emad M....P3.0 El-Sadr, Wafaa... P06.02, P06.03, P06.04, P23.4, P52.04 Else, Laura...OA27.0, OA27.06 LB Elsesser, Steven...OA07.06 LB Eluwa, George...P09.2, P49.04 Emerson, Cindi W....OA22.0 Emond, Mary J....PD02.0 Ende, Zachary...OA2.04 Ericsen, Adam...OA4.06 Ernandes, Michael J....PD05.02 Erra Díaz, Fernando...P24.02 Erraïs, Yasmine...P09.4 LB Escoda, Cristina...OA8.03 Espinoza, Lilia...OA3.02, P5.02, P23.7 Esteban, Mariano... P0.07, P4.05 Estes, Jacob D....OA7.05, P2.05 Estes, Jake D PL03.0 Estrada, Hernando...P2.5 Etima, Juliane...OA02.06 LB, OA09.0, OA5.02, OA5.04, P02.04, P23.0, P42.02 Etoa, François-Xavier P2.4 Etukoit, Bernard Michael...P52.08 Eudailey, J....OA06.02 LB Euler, Zelda...P34.2 LB Evans, Abbey...P4.0, P5.26 LB, P40.06 Evans, Tristan I....P2.09 Excler, Jean Louis...OA08.0, OA.05, OA2.06 LB, P26.3, P44.09 Exner, Theresa...P09.0 Ezeamama, Amara...P36.07 F Fadda, Elisa...PD05.0 Fahey, John V.... OA20.03, P5.24, P40.24, P40.25 Fairlie, Lee...RT02.04 Falivene, Juliana...PD05.04, P24.5, P4.07 Farah, Bashir... PD03.04 LB, P24.09, P26.04, P40.23 Farmer, Paul...OA2.0 Farrior, Jennifer...P52.04 Fashemi, Titilayo...OA0.05, P4.06 Fast, Pat...OA.0, P0.0, P.03, P3.4, P2.0, P26.03, P26.04, P36.0, P40.23, P49.08 Fast, Patricia...P26.07, P36.02, PD03.04 LB Fauci, Anthony S....PL0.03, OA7.02, P34.02 Fava, Joseph...PD04.02, P5.2 Fawzi, Wafaie...P36.07 Fazilleau, Nicolas...P2.08 Fedida, David...P33.06 Fedonin, Gennady G....P25.03 Felber, Barbara K....OA6.05, OA24.04, OA29.06 Feng, Yi...P39.0 Feng, Yu...PD03.03, P0.04 Feng, Zhimin...P2.5 Fennelly, Glenn...P4.08 Ferari, Guido...P26.08 Ferguson, Debbie H....P5.0 Feria, Manuel G....P04.02 Fernandez, Christine...P09.4 LB Fernandez, Natalia...P26.07 Fernandez, Stefan...P HIV Research for Prevention 204 HIV R4P

423 Author Index Fernández-Pineda, Alejandra...P5.04 Fernández-Romero, Jose...OA03.05, P4.02, P5.27 LB, P33.02 Ferrari, Guido...OA2.06 LB, OA25.0, P03.04 LB, P03.05 LB Ferreira, Thais...P29.05 LB Ferreira, Victor H....OA20.0 Fetherston, Susan...OA26.03, P4.03, P4.04, P4.06, P4.08 Feuer, Cindra... OA9.03, PD0.04 Feyaerts, Maxim...OA03.04 Fibla, Joan...P37.0 Fichorova, Raina...OA0.05, P25.0, P40.5, P4.06 Fidler, Sarah...OA28.03 Field, Sam...P3.5, P2.04, P48.06 Fields, Sheldon D....OA9.04 Fife, Kenneth...P49.0 Filali-Mouhim, Ali...OA04.05 LB Fillekes, Quirine...P22.03 Finnegan, Trisha...P23.6 Fischer, Will...OA2.06 LB Fisher, Kevin...PD06.02, P47.0, P47.02, P47.03, P47.07 Flach, Britta...OA.06 LB, OA6.06 Flamar, Anne-Laure...P4.5 Fletscher, James...OA05.04 Flingai, Seleeke...OA30.0 Fluharty, Tayler R....P03.0, P24.07 Flynn, Jacqueline K....P35.02 Fomsgaard, Anders... P2.07, P4.0 Fong, Youyi...OA4.03 Fong Lim, Michele...PD03.04 LB Fonteh, Pascaline...P5.02 Fontu, Alemju...P30.03 Forbes, Anna...P47.05 Forbes, Ben...P3.03, P44.2 LB Ford, Nathan...PD06.05 LB Ford, Susan...OA03.02 LB Forthal, Donald N....OA25.0, P03.04 LB Fouda, Genevieve...P03.05 LB Fouda, Pierre...P4.22 Foulds, Kathryn...OA25.0, P03.04 LB Foulger, Andrew...OA06.02 LB, OA2.06 LB Fourati, Slim...OA04.05 LB, OA25.0, P03.04 LB Fouts, Timothy R....OA24.04 Fowler, Mary Glenn... P05.0, P38.07, P49.05 Fox, Julie...OA07.03 Frahm, Nicole...OA04.05 LB, OA05.02, OA.06 LB, OA7.06, PD03.0 LB, P24.06, P26.4 Francella, Nicholas...PD03.02 Franchini, Genoveffa...OA04.04, OA06.03, OA25.0, P03.04 LB Francica, Joseph...OA6.04, P0.2 Francis, Donald...OA2.06 LB Francis, Suzanna...OA02.04, P40.2 Frank, Bruce...OA3.02, P4.05 Frank, Ian...OA05.02 Fraser, Kathryn...SY.02 Frater, John...OA4.04 LB Fredricks, David...P40.3 Freguia, Christian F....P44.02 Frezieres, Ron...P53.0 Friedland, Barbara OA02.0, P02.02, P09.6 LB, P5.27 LB, P49.03 Friedrich, Thomas...OA4.06 Friend, Chantel...P2.02 Friend, David...OA03.03, PD04.02, P5., P5.2, P5.6 Frohlich, Janet...OA9.06 Früh, Klaus...SY.03 Fuller, Deborah H....OA7.05 Fulton, Roice...P23.6 G Gabier, Ereshia...P34.04 Gadhe, Sharda...P4.26 Gaffoor, Zakir...P23.0, P53.03 Gafos, Mitzy OA02.04, OA07.03, P40.2 Galarraga, Omar...P06.0 Galaska Burzuk, Beth...OA3.05 LB Galea, Jerome T.... PD0.04 Gall, Astrid...P24.9, P39.03 Gallart, Teresa...OA8.03, P4.03 Gallo, Robert C....SY09.02 Gama, Cynthia...P50.0, P53.02 Gama, Lizzy... OA02.02, PD0.02 Gamble, Theresa... P06.02, P06.03, P06.04, P23.4, P52.04 Gamieldien, Hoyam...OA04.03, OA7.0, P3.03 Ganoza, Carmela...OA4.02, P24.08 Gao, Feng...OA2.06 LB Gao, Guofen...P0.08 Gao, Yajing... P5.7, P43.0 Garces, Fernando...OA25.04 Garcia, Felipe... P24.03, P27.0 Garcia, J. Victor...OA03.0, OA8.05 García, Elisabeth...P6.0, P6.04 García, Felipe...OA8.03, P4.03 García Pérez, Javier...OA22.05, P35.04, P44.04 García-Alonso, Dolores...P5.03 García-Arriaza, Juan...P0.07 García-Morales, Claudia...P22.02 Garrett, Nigel OA2.02, P39.05, P40.03, P40.09, P40.7, P40.2 Gartland, Andrew...OA08.04, P24.6 Garziano, Micaela. OA08.02, PD02.04 Gatell, Jose María...OA8.03, P4.03 Gati, Brenda M....P23.0, P38.07, P49.05 Gautam, Rajeev...OA30.05 Gazzard, Brian...PD03.04 LB Geffner, Jorge...P24.02 Geldmacher, Christof...P26.08 Geller, Elizabeth J....OA22.0 Gellerup, Dane...P0.06 Gelman, Marcy...OA09.06, P7.0 Gelmon, Larry...P9.0 Gengiah, Tanuja N....P3.05 Gentle, Nikki L....OA24.06 LB Georgia, Tomaras...OA.03 Georgiev, Ivelin S....OA0.06, OA08.04, OA30.05, P0.03, P0., P5.25 Geraghty, Daniel...OA4.03 Gerdts, Sarah...OA6.06 Getachew, Bethelihem... P05.08, P42.0 Gettie, Agegnehu...OA0.02 Ghebremichael, Musie...OA20.04, P40.08 Gherardi, María M....P24.5 Gherardi, Maria Magdalena...P4.07, PD05.04 Ghiglione, Yanina...PD05.04, P24.5, P4.07 Ghneim, Khader...P4.30 LB Ghosh, Mimi...P40.0, P40., P40.24 Gibbs, Anna...OA7.04, PD02.03 Gibbs, Richard...OA4.06 Gichuru, Evans...P07.08, P09.07, P3.06 Giguere, Rebecca...PD04.03, P42. Gilber, P....OA06.02 LB Gilbert, Peter...OA08.04, OA.03, OA.06 LB, OA4.03, PD03.0 LB, P26.4 Gile, Jillian...OA7.05, P2.05 Gill, Katherine...OA09.05, P53.02 Gillis, Jacqueline...P2.09 Gilmour, Christopher...P43.08 Gilmour, Jill...OA.0, OA4.0, OA2.0, OA2.03, PD03.04 LB, P24.09, P26.03, P26.04, P26.07 Gimour, Chris... OA26.03, P4.03, P4.06, P4.07, P4.08 Girard, Gabriel...P48.03, P48.05 Gita, Ramjee...P49.0 Githuka, George...P02.07, P2.04, P2.05, P48.06 Giudice, Linda...P40.28 Glashoff, Richard H P2.04, P2.2 Glaubius, Robert...OA27.05 Glenda, Ernst...P24.02 Glenn, Jolene A....OA0.05 Gluchowski, Marcel...P03.02 Gnudi, Federica...OA08.02, PD02.04 Godbole, Sheela...P09.08 Godeau, Jean Paul...P09.4 LB Godoy-Ramirez, Karina...OA.02 Goepfert, Paul...OA.03, OA4.0, OA2.0, OA2.03, P24.7, P26.7 Gofwen, Wika...P02.09 Gomez, Gabriela...P3.04 Gomez, Kailazarid...P42.03, P49.05 Gomez, Rafael...P33.05 Gómez, Carmen Elena...P0.07, P4.05 Gómez, Rafael...P5.0, P33.04 Gomez-Acebo, Eduardo... P4.02, P44.0 Gomez-Feliciano, Kailazarid...P49.06 Gonçalves, Emília...P26.6 Gonçalves, Thomas... P9.07 LB Gonelli, Christopher...P0.4 LB Gong, Tiantian...P5.06 Gonzales, Pedro... PD0.04 Gonzalez, Andrea...P06.0 Goode, Diana OA0.02 Gooneratne, Shayarana P25.0 Goonetilleke, Nilu...OA2.06 LB Goovaerts, Odin...P24.4 Gopalappa, Chaitra...P47.07 Gordon, Janna R....OA23.02 Gordon, Shari N...OA04.04, OA06.03, OA25.0, P03.04 LB Gorman, Jason... OA0.0, OA0.06 Gorny, Miroslaw K....OA05.06 LB Gorry, Paul R....P35.02 Gosse, Leslie...OA08.06 LB Goswami, Prabudhyagopal...P28.0 Goswami, Sandeep...OA0.04 Gottardo, Raphael...OA04.05 LB, OA08.04 Goujard, Cécile...P39.02 Goulder, Philip...OA04.02, OA4.04 LB, OA29.03, P2.7, P24., P24.9 Goulet, Jean-Philippe... OA04.06 Gounden, Natasha P38.0 Govender, Dhevium...P32.0 Govender, Rusha...P29.0 Govender, Vaneshree... P23.3, P32.0, P38.04, P53.03 Govender, Yashini...P05.09 LB, P33. LB Grab, Sheila...P5.20 Graebing, Philip...OA3.0 Graham, Barney...OA06.02 LB, OA7.06, PD03.0 LB, SY2.0 Graham, Susan M....P09.07, P3.06, P07.08, P3.6 LB Granich, Reuben...P47.0, PD04.04 Grant, Oliver C....PD05.0 Grant, Robert...OA09.06, P9.02 Grant, Shannon P....P24.08 Grasso, Chris...P7.0 Grau, Berta...P34.03, P34.06 Gray, Clive M....OA7.0, P3.03 Gray, Elin... OA2.02, OA30.03 Gray, Glenda...OA09.03, OA0.03, OA.04, OA.06 LB, OA9.05, P3.03, P23.08, P26.03, P26.2, P37.04, P38.03, P42.0, P47.06, P50.0, PD05.05, PL04.03 Gray, Lachlan R P35.02 Green, William...P0.03 LB Greenblatt, Ruth M...SY08.0, P40.28 Greene, Elizabeth...P06.02, P06.03, P06.04, P23.4, P52.04 Greene, Justin...OA4.06 Greene, Warner C....P25.05, P25.07 Greener, Ross...OA23.02 Greenspan, Suzan...P42.05 Gregor, Charles...P7.0 Grieco, Michael...P4.05 Griesbeck, Morgane...P2.6 Griffith, Sam...OA28.03 Grimm, Wesley A....P25.02 Grivel, Jean-Charled...OA30.06 LB Grobler, Anneke...P50.02 AUTHOR INDEX 423

424 Author Index AUTHOR INDEX Grooms-Williams, Tiffany N....P8.04 Grossman, Cynthia I....OA5.03, OA5.04, P42.04 Grove, Doug...P26.09 Grunenberg, Nicole...OA.03, OA.06 LB Gu, Niya...OA.06 LB Guan, Yongjun... OA24.04, OA30.02 Guardo, Alberto C....P27.0 Gudo, Elias...P09.0 Guenaga, Javier...OA0.02, P0.04 Gueye, Daouda...P3.8 LB Gugasyan, Raffi...P40.29 Guigo, Roderic...OA24.06 LB Gumbe, Anne...PD03.04 LB Gumbi, Pamela P....OA04.03, P40.07, P40.4 Gumede, Siphiwe...P38.03 Gundacker, Holly M....P38.07 Gunn, Bronwyn...P6.07 Gupta, Gopal...P44.0 Gupta, Pankaj...P39.09 Gupta, Phalguni...P5.06, P40.27 Gupta, Somya... PD04.04 Gurley, Thaddeus C....OA06.02 LB, OA2.06 LB Gurwith, Marc...OA6.0 Guwatudde, David...P36.07 H Haase, Ashley...SY.02 Haazen, Wouter...P5.09 Haberer, Jessica...OA07.06 LB, OA28.0, P43. Haberland, Nicole A....P29.06 LB Haddad, Elias...OA29.0 Haddad, Lisa...OA20.06 Haigwood, Nancy L....OA2.03 Haire, Bridget G....PL02.02 Hallett, Timothy B....P20.0 Ham, Anthony...P5., P5.2, P5.9, P5.23, P44.02 Ham, Claire...P5.0 Hamanishi, Kristen A....OA27.0 Hamm, Tiffany...P23.02 Hammer, Scott...OA06.02 LB, PD03.0 LB, P07.02, P26.09 Hammond, Katherine B....SY.03 Hamorsky, Krystal...P44.06 Hancock, Gemma.... OA05.05, P26. Hanke, Tomas...OA29.06, P26.05, P26. Hannah, Stacey...P07.03, P07.04 Hannaman, Drew...P26.07 Hansen, Scott G....SY.03 Hao, Yanling...P4.6 Hapgood, Janet P....OA20.02 LB, P05.09 LB, P33. LB Haq, Kamaran...OA6.03 Hara, Hiroto...PD03.04 LB Harada, Shigeyoshi...P35.03 Harbolick, Elizabeth...OA2.06 LB, P25. Hardy, Liselotte...P40.5 Hari Kumar, R...P36.05 Haribhai, Kshama...P.02 Harichund, Charlene...P23.06 Haridutt, Avika...P26.0, P38.05 Harit, Dimple...P40.6 Harmon, Thomas...P47.0, P47.07 Harrigan, P. Richard...P37.03 Harris, Marianne... OA04.06 Harris, Max...P0.06 Harrison, Polly...P47.0 Hartley, Oliver...P35.04 Hartling, Hans Jakob...P2.07 Hartmann, Miriam...OA02.05, OA02.06 LB, OA5.02, OA5.03, OA5.04, P42.04, P46.03, PD04.0 Hasegawa, Mamoru...PD03.04 LB Hassan, Namir...OA05.05 Hatzenbuehler, Mark...OA07.04 Hauber, Joachim...P4.28 Haugh-Krumpe, Lauren...P33.0 Haule, A....P26.08 Hay, Christine...OA05.02, OA.03 Haydont, Christine...P09.4 LB Hayes, Peter... PD03.04 LB, P24.09, P26.03, P26.07 Hayes, Richard...OA5.0, OA28.03, P40.2 Haynes, Barton F....OA06.02 LB, OA2.06 LB Hayton, Emma-Jo...P26. Hazarika, Anjali...P2.0 Hazra, Rohan...P45.05 He, Cui...P39.0 He, Xiang...P39.0 Headley, Jennifer...P3.5, P46.0 Hearps, Anna...P40.29 Heath, Sonya...OA05.03, P24.7 Heckerman, David...OA4.04 LB, OA2.0 Heeres, Jan...P35.0 Heffron, Renee...OA27.02, OA28.0, OA28.02, P46.06 Heit, Antje...OA6.06 Hejdeman, Bo...OA.02 Helgadóttir, Berglind...P5.4 Henderson, Marcus H....OA22.04 Hendrix, Craig W....OA07.06 LB, OA3.0, OA3.02, OA3.05 LB, P8.06 Heneine, Walid...OA03.06 LB Hengprasert, Sumetha...P36. Henn, Matthew R....OA2.05 Henrick, Bethany M.... P2.3, P40.26 Hensley-Mcbain, Tiffany... OA7.05, P2.05 Herbst, Kobus...SY03.03 Herman, Colleen...P09.8 LB, P9.05, P23.04 Herman, Melissa M....P24.3 Hermanus, Tandile...P4.26 Herold, Betsy C....OA0.05, OA3.02, OA3.05 LB, P5.02, P8.03, P23.7, P44. SY0.02, OA3.03, P49.4 Herr, Christopher...P25. Herrera, Carolina...OA08.0, OA22.05, P2.06, P5.26 LB, P35.04, P44.04 Herrick, Amy...OA27.06 LB, P23.03 Herschhorn, Alon...P5.28 LB Herst, C V....P4. Hertz, Tomer...OA08.04, P24.6 Hessell, Ann J....OA2.03 Hewett, Paul C....P29.06 LB Heydarchi, Behnaz...P0.4 LB Heyndrickx, Leo...P35.0, P39.03, P4.0 Hiatt, Joseph...OA7.02, P34.02 Hickman, Taylor...OA29.0 Higgs, Chris...OA07.03 Hijazi, Karolin...P3.0, P3.02 Hilda, Shumba P....P09.05 Hill, Brenna...OA06.0 Hillier, Sharon L....OA0.0, OA0.06 LB, OA3.0, P5.07, P8.06, OA20.05 LB, OA22.03, P49.07, P53.04 Hilton, Joan...P40.28 Hirata, Izumi...P34.05 Hirbod, Taha...OA7.04, PD02.03 Hironaka, Takashi...PD03.04 LB Hladik, Florian...P40.04 Ho, David...SY2.02, P39.02 Ho, John...P37.08 Hoelscher, Michael...P26.08 Hoesley, Craig... P5.27 LB, P53.04 Hoffman, Irving...P4.0 Hoffman, Susie...P09.0 Hoffner, Michelle...OA2.05 Hogan, Michael...P4.25 Hold, Georgina L....P3.0, P3.02 Holding, Jeremy...P8.05 Holgado, Maria Pia...P4.07 Holt, Jonathon...OA26.03, P4.0, P4.03, P4.04, P4.05, P4.06, P4.07, P4.08, P8.05 Holtz, Timothy H.... OA07.05, P09.3, P3., P30.02, P36., P49.2 Homan, Rick...P50.06 LB Hong, Christin M....OA05.0 Hong, Heather A....P37.04 Hong, Kunxue...P6.02 Hood, Greg...OA27.05 Hope, Thomas J....OA04.04, OA7.0, P3.03, P6.07, P25.02, P40.05 Hopewell, Nicola...OA08.06 LB Hornschuh, Stefanie...OA09.03 Hosek, Sybil...RT04.04, P9.02 Howell, Shana...P25. Hoxie, James A....P4.25 Hsi, Jenny H....P39.0 Hsueh, Jessica...OA0.03 Hu, Minlu...OA22.03 Hu, Xintao... OA6.05, OA29.06 Hu, Yuanyuan...P6.02 Hua, Yuanzi... OA0.03, OA25.04 Huang, Jinghe...OA6.0 Huang, Po-Ssu...OA25.05 Huang, Ying...OA.06 LB Huang, Yunda...P26.4, P39.0 Huber, Ashley...OA0.05, OA3.02, P44. Hughes, Collette M....SY.03 Hughes, Lindsay... P23.02, P3.8 LB Hulsman, Mechtild...P09.5 LB Hume, Steve...P34.0 Humeau, Laurent M....P26.5 Hungu, Charity...P22.05 Hunt, Hazel...P05.09 LB, P33. LB Hunt, Peter W.... P37.03, P43. Hunter, Eric...RT04.05, OA08.05, OA.0, OA4.0, OA4.04 LB, OA2.0, OA2.03, OA2.04, OA28.04, P0.0, P3.02, P3.4, P24.2, P24.7, P26.7, P39. LB, P52.0 Hunter, Meredith...OA24.0 Husk, Adam...P44.06 Husnik, Marla...P0.02 Hutchinson, Craig S....OA9.04 Hutnick, Natalie...OA24.0, P0.0 I Ibba, Salomè... OA08.02, PD02.04 Ibitamuno, Grace...P25. Ibitoye, Mobolaji... PD04.03 Idoko, John... OA28.05, PD0.03 Ifekandu, Chiedu C.... P3.07, P3.08 Ighil, Julien...P2.0, P40.20 Ighodaro, Michael...OA9.03 Ikaneng, T...P23.08 Inambao, Mubiana...RT04.05, P08.0, P09.0, P3.0, P3.4, P23.6, P36.06, P42.0, P52.0 Indangasi, Jackton...P24.09, P40.23 Ingabire, Rosine... PD03.04 LB, P05.03, P3.02, P40.22, P49.08 Ingale, Jidnyasa...PD03.03 Ingalls, Robin R....P40.02 Innes, Craig...OA.06 LB Inoue, Makoto...PD03.04 LB Introini, Andrea...OA7.04 Ipp, Hayley...P2.04, P2.2 Iriaye, Desmond A....P49.04 Irungu, Elizabeth... OA27.02, OA28.02, P46.05 Irwin, Juan...P40.28 Isaacs, Karen...P45.07, P50.04 Isaacs, Michelle...OA09.02, P9.05, P23.04, P53.02 Isac, Shajy...P20.02 Ishii, Hiroshi...P4.20 Islam, Ayesha...P40.02, P4.9 Ismail, Arshad...P34.08 Ismail, Nasreen...OA29.02, OA29.03, P24.0, P24.2 Ismail, Zaheda...P38.03 Iwajomo, Oluwadamilola H...OA6.03 Iwuji, Collins...SY03.02, P52.05 Izquierdo-Useros, Nuria...P24. J Jackson, Akil...OA27.0, PD03.04 LB Jackson, Ed...P48.03, P HIV Research for Prevention 204 HIV R4P

425 Author Index Jackson, Suzanne S....P40.30 Jacob, Rajesh A P34.06, P34.03 Jacobs, Bertram L...P0.07 Jacobs, Lisa...P43.05 Jacobs, Jr., William...P4.08 Jacobson, Cindy...P23.03, P45.07 Jacot, Terry A....P23.5 Jadhav, Praveen...P3.09 Jaeger, Frederick...P0.2 Jagesur, Lakshmi... P38.05, P38.06, P39.04 Jaggernath, Manjeetha...P2.7 Jain, Sachin...P7.02 Jais, Mariel...P40.0, P40. Jakobsen, Bent K....OA05.05 Jalah, Rashmi...OA24.04 Jamieson, Denise P03.05 LB Jamshidi, Roxanne...P23.5 Jana, Smarajit...OA27.04 Jani, Ilesh...P26.6 Jansson, Marianne...P03.03 LB Jaoko, Walter...PD02.05 Jaspan, Heather B....OA04.03, OA7.0, P3.03, P40.07 Jaumdally, Shameem Z....OA04.03, OA7.0, P3.03, P40.4 Jean-Pierre, Ninochka... P4.02, P33.02 Jeenarain, Nitesha...P53.03 Jelicic, Katija...OA7.02, P34.02 Jennes, Wim...P24.4 Jensch, Ute...P40.2 Jensen, Kara...P4.08 Jensen, Sanne S....P4.0, P2.07 Jenson, Alfred...P8.04 Jespers, Vicky...P40.5 Jessen, Heiko...OA2.05 Jia, Manhong...P39.0 Jiang, Hao...PD03.03 Jiang, Shibo...P25.05 Jiang, Xunqing...P0.3 LB Jiang, Yonghou...OA26.05, P5.8 Jimenez, Esther...P24. Jiménez, Jose Luis... P5.0, P33.05, P5.04 Jiménez, Victoria...P0.07 Joachim, Agricola...P26.08 Joag, Vineet R....OA7.03 Jobe, Ousman...P0.08 John, Mina...OA4.04 LB John-Langba, Johannes...P42.07 Johnson, Brent...OA20.06 Johnson, Brian...OA7.05 Johnson, Brittni...P40.28 Johnson, Cheryl...P09.04, P09.5 LB Johnson, Sarah A....P24.06 Johnson, Susan...SY.04 Johnson, Todd...OA03.03, P5.6 John-Stewart, Grace...P22.0 Joliot-Vilain, Mireille...P09.4 LB Jollimore, Jody...P48.03, P48.05 Jones, Bryn...P4.3 Jones, Leonard P0.03 LB Jongrakthaitae, Surat...OA05.04 Jongwe, Tsungai I....P4.09 Joossens, Jurgen...P35.0 Joseph, Fava...P5. Joseph, Sarah... P26.08, P40.2, P40.5 Joshi, Paulatsya...P26.06 Jost, Stephanie...OA04.0 Joyce, Gordon...P5.2 Joyce, M. Gordon...OA0.06 OA06.06, P0.03, P0. Juarez-Figueroa, Luis P06.0 Jubb, Becky...P35.02 Juggernath, Vermala...P05.07 Julien, Christophe...P09.4 LB Julien, Jean-Philippe...OA0.03, OA25.04 Juraska, Michal... OA08.04 K Kaale, Anna...P3.5 Kaaya, Sylvia...P3.5 Kabakyenga, Jerome...OA23.03 Kabarambi, Anita...P49. Kabwigu, Samuel...OA09.0, OA5.02, P02.04, P05.04, P23.09, P23.0, P38.07, P42.02, P45.05, P45.07 Kadam, Ravindra...P4.0 Kadasia, Kadryn...OA24.02 Kaewkungwal, Jaranit...OA04.05 LB, OA08.0, OA08.04, OA2.06 LB, OA4.03, P26.4, P44.09 Kafka, Jessica K....OA20.0 Kaggwa, Mary...P23.09 Kaggwa- Katumba, Angelo... P43.05, P43.09 Kaguiri, Eunice...P49.0 Kaida, Angela... OA09.03, OA23.03 Kakayi, Brenda...P49.06 Kalams, Spyros...OA2.03 Kale, Kavita...P40.8 Kaleebu, Pontiano...PL02.03 Kalibbala, Margaret...P05.05 Kalil, Jorge...P4.04 Kamali, Anatoli...OA02.04, OA.0, P0.0, P05.05, P09., P.04, P2.0, P36.0, P36.02, P49.02, P49., P53.02 Kamarebe, Josephine...P43.2 Kamgaing, Rachel...P2., P2.4 Kamira, Betty...P05.04, P38.07, P49.05 Kamusoko, Miriam... P09.05, P48.02 Kamwendo, Deborah...P03.05 LB Kamya, Philomena...OA29.02 Kamya Nsangi, Justine...P05.04 Kang, Byong...OA6.0 Kanyemba, Brian...OA9.03, PD0.04, PD04.05 Kanyiki, Leader... OA09.05, OA9.05 Kapaata, Fred B....P05.04, P23.09 Kapewangolo, Petrina...P5.02 Kapiga, Saidi...P40.2, P53.02 Karabarinde, Alex...P49.02 Karamov, Eduard...P33.03 Karas, Sherri...P23.8 Karasavvas, Nicos...OA.05, P26.3, P44.09 Kariko, Katalin...P4.25 Karim, Abdool S....OA04.02 Karita, Etienne...OA.0, OA28.04, PD03.04 LB, P0.0, P05.03, P09.0, P3.02, P24.2, P26.07, P26.2, P40.22, P49.08 Kariuki, Thomas M....P25.06 Karl, Julie...OA4.06 Karlsson, Ingrid...P2.07, P4.0 Karlsson Hedestam, Gunilla B.. SY02.02 Karnasuta, Chitraporn...OA.05 Karuna, Shelly T... OA9.0, PD03.0 LB, P07.02, P24.08, P26.09 Karuppiah, Muthumani...P0.0 Kasamba, Ivan...P49.02 Kasango, Christian...P38.03 Kashuba, Angela D.M....OA3.03, OA3.04, OA22.0, OA22.04, OA22.05, OA22.06 LB, OA26.06 LB, P5.24 Kaslow, Richard...OA2.0 Kasprowicz, Victoria...OA04.02 Kastner, Jasmine...OA23.03 Kat, Pizzoferro...P26.06 Katabira, Elly...OA28.0, OA28.02, P24.07 Katinger, Dietmar...P26.06 Katz, David F....OA22.04, PD04.02, P5., P5.2, P5.7, P5.9, P43.0 Katzen, Lauren P49.03 Kauffman, Daniel...OA29.0, OA29.04 Kaul, Rupert... OA7.03, PD02.03, P40.9 Kaushic, Charu...OA20.0 Keefer, M....OA06.02 LB Keele, Brandon F.... OA7.05, OA25.0, P03.04 LB, P2.05 Kehrl, John...P34.02 Kellam, Paul...OA06.0, P24.9, P39.03 Kelleher, Neil...P6.07 Keller, Marla J....OA3.02, P5.02, P23.7, P49.4 Keller, Tibor...P4.22 Kelly, Charles...P3.03, P35.04, P44.04, P44.2 LB Kelly, Christine A....P29.06 LB Kelly, Cliff...OA0.06 LB, OA3.05 LB, P42.05, P49.06 Kelly, Elizabeth P....P24.06 Kelsoe, Garnett...SY0.04 Kelvin, Elizabeth...P09.0 Kemigisha, Doreen...OA09.0, P02.04, P23.0 Kene, Terfa...P09.2 Kenney, Jessica...OA03.05 Kent, Stephen...P25.0, SY06.04 Kepler, Thomas...OA06.02 LB, OA2.06 LB, OA6.04 Keshinro, Babajide...P23.02 Keshinro, Jide...P09.2 Kestens, Luc...P24.4 Khamadi, Samoel...P23.02 Khan, Amir...OA24.0, P26.02 Khanukkova, Elena...P23.03 Kharsany, Ayesha...P.02 Khasimwa, Brian...P22.0 Khati, Makobetsa...P5.5 Khunwane, Mamakiri...P23.08 Kiarie, James...P20.03 Kiazyk, Sandra A.... P24.04, P24.05, P24.3 Kibengo, Freddie M.... P2.0, P36.0, P36.02 Kibler, Karen V....P0.07 Kibuuka, Hannah...OA2.06 LB, P23.02, P26.02 Kidane, Segen...OA7.03 Kidd, Val...P53.02 Kidoguchi, Lara...OA28.0 Kiio, Maria...P25.06 Kijak, Gustavo...OA4.03, OA2.06 LB, P25., P39.07 Kikonyogo, Florence...P23.09 Kilbourne-Brook, Maggie...OA26.0 Kilembe, William...RT04.05, OA.0, OA4.0, OA20.06, OA2.0, OA2.03, OA2.04, P0.0, P05.08, P08.0, P09.0, P09.05, P3.0, P3.4, P23.6, P26.03, P26.07, P26.2, P36.06, P39. LB, P42.0, P48.02, P52.0 Kilgore, Katie...OA08.05, PD03.02 Killingbeck, Sarah S....P4. Kilonzo, Nduku...RT0.04 Kim, Helen J.... OA25.04, OA25.05 Kim, Jerome H....OA04.05 LB, OA05.04, OA08.0, OA08.04, OA.05, OA.06 LB, OA2.06 LB, OA4.03, OA2.06 LB, OA23.0, OA25.0, P03.04 LB, P25., P26.3, P26.4, P39.07P44.09 Kim, Joseph J....OA30.0 Kim, Kyeong-Ae...P25.05 Kimani, Joshua...OA08.03, OA7.03, OA7.04, PD02.03, PD02.05, P03.02, P9.0, P24.04, P24.05, P24.3, P37.02, P37.08, P40.9, P40.3 Kimani, Makubo...OA08.03, P24.04, P24.05, P40.3 Kimaru, Linda...P3.0, P36.06, P52.0 Kimble, Thomas D....OA0.05, P23.5 Kimpel, Janine...P4.2 Kimulwo, Maureen J.... P22.05, P25.04 Kimwaki, Steve...OA7.03 King, Caroline...OA0.04, P03.05 LB, P34.0 King, Deborah F.L....OA24.05, OA30.06 LB, P27.03, P40.06, P4.3 King, Elizabeth...P3.8 LB King, Georgette...OA9.04 Kinloch, Natalie...P37.03 Kintu, Kenneth P45.05 Kinyua, Joyceline...P49.09 Kirchhoff, Frank...P25.05, P25.07 Kireev, Dmitry E....P25.03 AUTHOR INDEX 425

426 Author Index AUTHOR INDEX Kirunda, Kakaire A....P43.09 Kirys, Tatsiana... OA08.04 Kiser, Patrick...OA03.03, OA04.04, OA3.02, OA26.02, P5.6 Kishore, Uday...P40.8 Kitawi, Rose C....P22.05, P25.04 Kiwanuka, Noah...P05.06, P23.07 Kiweewa, Flavia Matovu...OA09.0, P23.09, P42.02, P42.05, P38.07, P49.05,P53.03 Kiweewa, Francis...P23.02 Kizima, Larisa... OA03.05, P4.02, P33.02 Klatt, Nichole R....OA7.05, P2.05, P40.3 Klatzmann, David...P27.05 Klausner, Jeffrey D....P08.02 Klein, Daniel J....P43.5 LB Klein, Florian...P6.03, P34.09 Klein, Katja...P40.06 Kleinbeck, Kyle...P5.27 LB, P33.02 Kliche, Alexander...P4.27 Klindera, Kent...OA9.02 Kloverpis, Henrik...OA04.02, P24. Kluge, Silvia...P25.05 Klungthong, Chonticha...P25.08 Kobinger, Gary... P4.7, P4.8 Kochar, Nidhi...OA05.02 Koday, Michael... OA7.05, P2.05, P40.3 Kohli, Rewa...P3.09 Kojic, Erna M....PD04.02, P5., P5.2 Kolawole, Grace O....PD0.03 Kombo, Bernadette...P07.08, P3.6 LB Konda, Kelika...P08.02 Kong, Leopold... OA25.04, OA25.05 Kong, Wing-Pui...P0.03 Kong, Xiang-Peng...OA05.06 LB, P0.3 LB, P26.8 Kongpechsatit, Oranuch...P25.08 Koofhethile, Catherine K....OA29.03 Kopycinski, Jakob...OA4.0, OA2.03 Korber, Bette...OA2.06 LB Korner, Christian...P2.03 Kornilaeva, Galina...P33.03 Kosakovsky-Pond, Sergei L...OA2.02 Kosgei, Josphat...P26.02 Kotlewski, Jennifer A....P3.0 Kotze, Philip...P2.02, P53.02 Kouassi, Pascale... OA04.06 Kouokam, Joseph...P8.04 Koup, Richard...OA06.02 LB, OA06.0, OA6.0, OA25.0, OA30.05, P03.04 LB,P4.30 LB Kourjian, Georgio...P24.20 LB Kourtis, Athena...P03.05 LB Koutsoukos, Marguerite...SY09.03 Kozlowski, Pamela...P4.08 Krakower, Douglas... OA07.04, PD06.04 LB Kramski, Marit.....P0.4 LB, P25.0 Kraus, Guenter...OA03.0 Krebs, Shelly...OA23.0 Kreppel, Florian...P4.2 Kriek, Jean-Mari...P40.07, P40.4 Krishna, Atul...P44.0 Krishna, Shagun...P44.0 Kroidl, Arne...P26.02, P26.08 Kroon, Eugene...OA05.04 Kruger, Sieglinde...P23.04 Krykbaeva, Marina...OA24.03 Kubeka, Muriel... OA23.04, OA26.0 Kublin, James...OA0.03, OA.04, PD06.0, P42.0 Kuhn, Louise...RT02.03, PD05.05, P37.04 Kuhn, Warren...OA04.02 Kulane, Asli...P4.2 Kulkarni, Viraj...OA6.05, OA24.04, OA29.06 Kulp, Dan...OA2.04 Kumar, Abhinav... P3.03, P44.2 LB Kumar, Rajesh...P2.0 Kumar, Sanjeev...P2.0 Kumar, Selva...OA8.0 Kumwenda, Jacob...P03.05 LB Kuo, Caroline...P06.0 Kusasira, Adrine...OA23.03 Kusemererwa, Sylvia... P05.05, P49. Kushwaha, Bhavana...P44.0 Kutzler, Michele...OA24.0 Kuwata, Takeo...P34.05 Kwa, Suefen...OA05.03 Kwatampora, Jessie P40.9 Kwena, Zachary...P30.04, P48.04 Kwok, Cynthia...P25.0 Kwon, Douglas S....OA20.04, OA24.03, SY08.02, P2.03, P40.08 Kwon, Young Do...P5.25 Kwong, Peter D.. OA0.0, OA0.06, OA06.06, OA08.04, OA2.0, P0.02, P0.03, P0., P5.2, P5.25, P5.28 LB, P39.02, SY05.03 Kyeyune, Rachel...P36.07 Kyomugisha, Juliet...P05.05 Kyongo, Jordan K....P40.5 L La, David...P37.08, P4.7, P4.8 Labbett, Rita Lisa...P23.03, P23.8 Laborde, Nicole... PD04.0 Labranche, Celia...P0.0 Lacabaratz, Christine... P24.0, P4.5 Lacap, Philip... P4.7, P4.8 Lacey, Charles...OA07.03 Lachau-Durant, Sophie...OA03.0 Ladnaya, Natalia N....P52.07 Lagat, Nancy...P49.09 Lagatie, Ole...OA03.04 Lagenaur, Laurel...OA8.04 Laher, Fatima...OA.06 LB, P24.0 Lai, Jaim J....OA3.03 Lai, Samuel...P40.6 Lakhi, Shabir...OA4.0, OA2.0, OA2.03, P08.0 Lakouga, Howard Y.... OA04.04 Lama, Javier...OA4.02, P24.08, P42., P52.03 Lambson, Bronwen...OA2.02, P34.08 Lamothe, Pedro...OA29.04 Landais, Elise...OA.0, OA2.02 Langa, Nonhlanhla...P23.06 Langat, Robert... P24.09, P26.04, P40.23 Lanham, Michele...OA02.0 Laplana, Marina...P37.0 Larmarange, Joseph...P52.05 Larsen, Brendan B.... OA08.04 Larsen, Michelle...P4.08 Larsen, Tine K....P2.07 Larson, Heidi...P48.07 Lassauniere, Ria...PD05.05, P37.05 Latham, Catherine...P40.0 Lauck, Michael...OA4.06 Laufer, Dagna... PD03.04 LB, P26.03, P26.04, P26.2, P40.22, P40.23 Laufer, Natalia...PD05.04, P24.5 Lauffenburger, Douglas...OA0.04, OA2.05, P40.9 Laumond, Géraldine... P6.05, P6.08 Launay, Odile...P26.0 Lawson, Benton...OA2.03 Lazzaro, Michelle...P25. Le Breton, Anne...P2.0, P40.20 Le Gall, Jean-Marie... P9.07 LB Le Gall, Sylvie...OA6.05, P24.20 LB Le Grand, Roger... SY02.0, OA03.04, OA08.06 LB, P 27.02, P44.05 Lebina, Limakatso...P29.02, P29.03 LeBlanc, Marc-André..P9.04, P48.03, P48.05 Lederle, Alexandre P6.08 Lee, Benhur...P35.02 Lee, Boon Kiat...P4.4 Lee, Carter...OA.06 LB, OA2.06 LB Lee, Guinevere Q....P37.03 Lee, Jenica L....P25., P39.07 Lee, Peter S.... OA0.03, OA8.04 Leeansyah, Edwin...OA7.04 Leelawiwat, Wanna...P25.08 Lees, Shelley...P48.07 Lefebvre, Francois...OA04.05 LB Legasse, Alfred W....SY.03 Leggett, Adam...P8.05 Lehman, Dara...P22.0 Lehrman, Jennifer...P26.04 Leider, Jason...P06.03 Lelièvre, Jean-Daniel...P26.0 Lembethe, Duduzile...OA9.05 Lemons, Ansley...P.0 Lemos, Maria P....P24.08 Lempicki, Richard...P34.02 Lenicov, Federico Remes...P24.02 Lennon, Niall J....OA2.05 Lenzi, Rachel...OA02.0 Leon, Segundo...P08.02 Leonavicius, Karolis OA25.05 Lépine, Aurélia...OA28.05 Lepore, Steven...P25. Lert, France...P52.05 Lertpruek, Sirirat...P3. Leslie, Alasdair...OA04.02 Leuvennink, Mildie...P53.0 Levendosky, Keith..P5.27 LB, P33.02 Levin, Carol...P09.7 LB Levin, Clement...P2.02, P2.08 Levin, Jonathan...P36.0 Leviyang, Sivan...OA2.06 LB Levy, Lisa... OA3.05 LB, OA5.03, OA5.04, P0.02, P42.04 Lévy, Yves...P24.0, P26.0, P4.5 Lewin, Sharon R....P35.02 Lewis, David... OA7.0, P3.03, P26.06, P40.07 Lewis, George K.... SY09.02, OA25.03, OA30.02 Lewis Kulzer, Jayne...P36.08, P45.08 Li, Dongliang...P29.04 LB Li, Haiying...OA04.0, P2.09 Li, Hualin...OA04.0 Li, Hui...OA2.06 LB Li, Jing...P5.05 Li, Jingjing...OA6.02, P6.09 Li, Jinyao...OA24.04 Li, Qingsheng... P4.7, P4.8 Li, Sue...OA05.02 Li, Wei...P0.3 LB Li, Zhenpeng...P6.02 Liang, Ben... P4.7, P4.8 Liang, Binhu...P37.08 Liang, Frank...OA25.0, P03.04 LB Liao, Hua-Xin...OA2.06 LB Liao, Larry...OA06.02 LB, P03.05 LB Liao, Lingjie...P39.0 Licht, Anna...PD05.03 Lichterfeld, Mathias...OA29.0 Lie, Yolanda...OA2.02 Liebenberg, Lenine...P40.03, P40.7, P40.9, P40.2 Lieberman, Judy...OA4.05, P33.09 Lifson, Jeffrey...P4.08, SY02.03 Lihana, Raphael...P49.09 Likhitwonnawut, Udom...RT04.02, PD0.04, P07.03 Lindegger, Graham C.... P.03, P.05 Lindqvist, Madelene...OA29.0 Lingappa, Jairam R....OA0.04, PD02.0, P03.0, P24.07 Lingping, Cai...P43.2 Linton, Christine...P0.09 Lioux, Thierry... P27.04, P4.03 Lisanti, Antonella...P2.03 Lissom, Abel...P2. Little, Francesca...OA04.03, P40.4 Little, Susan J....PD05.03 Liu, Albert...P9.02 Liu, Chang...P0.05, P4.6 Liu, Haichuan...P HIV Research for Prevention 204 HIV R4P

427 Author Index Liu, Karen...P42.03 Liu, Qingsheng OA6.02 Liu, Ying...P0.05, P4.6 Liu, Yu...P29.04 LB Liu, Zheng...P0.05 Livant, Edward...OA0.06 LB, P49.05 Livrozet, Jean-Michel...P09.4 LB Liyanage, Namal P.M....OA04.04, OA25.0, P03.04 LB Liynage, Namal...OA06.03 Llano, Anuska...OA29.06 Lloyd, K.E....OA06.02 LB Lloyd, Krissey E....OA2.06 LB Lloyd, Sarah B....P25.0 Lo Caputo, Sergio...OA08.02, PD02.04 Loescher, Thomas...P36.07 Logan, Murray G....P39.05 Lombard, Carl...P50.0 Lombardo, Angela...PD03.04 LB London, Grace M....P5.5 Loots, Stanley...P2.04 Lopatukhin, Alexey E....P25.03 Lopez, Ana...P33.05, P5.03 Lopez Rios, Javier...P3.7 LB Lore, Karin...OA25.0, P03.04 LB Lorente, Raquel...P5.0 Lorenzana de Rivera, Ivette...P22.02 Lortat-Jacob, Hugues...P44.05 Lou, Qi...OA6.02 Louder, Mark...PD05.02, P0., P5.28 LB Louw, Cheryl E....OA09.02, P9.05, P23.04, P53.02 Loxley, Andrew...P4.05 Lozano, Francisco...OA8.03 Lu, Richard...SY.04 Lu, Shan...P0.3 LB, P26.8 Lu, Shiqiang...P6.09 Lucas, Jonathan P.... OA07.0, OA9.04, PD0.04 Lucas, Judith...P6.07 Luechai, Pikunchai... OA07.05, P09.3, P49.2 Luhthuli, Faith Smangele...OA23.04 Lumngwena, Evelyn N....P39.06 Lump, Edina...P44.07 Lund, Jennifer M....P03.0, P24.07 Luo, Ma...OA08.03, P37.08, P4.7, P4.8 Luongo, Timothy S....P0. Lusti-Narasimhan, Manjula...P5.4 Luthra, Kalpana...P2.0 Luthuli, Faith...OA23.02 Luthuli, Londiwe...OA9.06 Lwembe, Raphael M....P22.05 Lyamuya, Eligius F....P26.08 Lynch, Rebecca M....OA30.05, P0.2 M Ma, Liying...P6.02 Ma, Yanling...P39.0 Ma, Zhong-Min...OA25.0, P03.04 LB Maarschalk, Silvia...P.02 Maboa, Rebone...P50.0 Maboko, Leonard...P26.02, P26.08 Mabota, Norma...P26.6 Mabude, Zonke...OA9.05 Mabuka, Jennifer K....P34.2 LB Macdonald, Kelly S....OA6.03 Macharia, Gladys OA.0, OA4.0, OA2.03 Macio, Ingrid S....OA0.0 Mackelprang, Romel D....PD02.0 Mackenzie, Caroline...P02.07, P2.04, P2.05, P48.06 Mackenzie, Charlene...P0.4 LB Mackie, Nicola E....OA07.03 MacLeod, Daniel T....OA2.02 MacMillan, Daniel...P37.03 Macovela, Eulália...P26.6 MacQueen, Kathleen M....OA02.03, P50.03 Madan, Taruna...P40.8 Madani, Navid...P5.28 LB Madanick, Ryan D....OA22.06 LB Madiga, Maphuti...OA30.03 Madnote, Sirinan...OA.05 Maduna, Nomfundo...P38.03 Maenza, Janine...OA06.02 LB, OA7.06 Maeto, Cynthia...P4.07 Maganga, Lucas OA2.06 LB, P23.02, P25. Magaret, Craig A.... OA08.04 Magidson, Jessica F....P08.03 Magnuson, David...OA5.06 LB Mahaka, Imelda...OA02.05, OA5.03 Mahal, Lara K....P49.07 Mahalingam, S...P39.09 Mahan, Alison E....OA25.02 Maharaj, Rashika...P26.0, P38.0, P38.05, P38.06, P39.04 Mahomed, Mehebub...P29.05 LB Mahon, Tara...OA05.05 Mahungela, Sannie...P24.0 Mainkar, Mandar...P28.0 Majola, Nelisile... P40.03, P40.09, P40.7 Major, Ian...OA26.03, P4.08 Majors, Alesha...OA02.03 Makgopa, Sylvia...P23.08 Makhanini, Bonus...P53.03 Makhdoomi, Muzamil A....P2.0 Makumbi, Fredrick...P05.06 Makwaga, Olipher...P45.06 Malahleha, Mookho...P2.03 Malamatsho, Ross...OA9.05 Malcolm, Karl...OA03.04, OA22.05, OA26.03, P4.03, P4.04, P4.06, P4.07, P4.08, P5.4 Malefo, Matshidiso...P23.08 Malherbe, Delphine...OA2.03 Mallal, Simon...OA4.04 LB Malone, Stephanie...OA22.06 LB Manentsa, Mmatsie...P29.03 Manenzhe, K...P23.08 Mango, Thabiso...P09.6 LB Mann, Brendan T....P39.07 Mann, Jamie F.S.... P27.03, P4.3 Mann, Philip...P26.08 Manning, Judy M....RT05.0 Manso, Bryce A....PD03.0 LB Mansoor, Leila...OA9.06, P23.06, P50.05, P50.06 LB Mantell, Joanne...P09.0 Manthata, Goitse...P23.05, P46.04 Mapfumo, Rumbidzai...P43.2 Maphumulo, Busi V....P30.06 Maphumulo, Lungi...OA29.03 Mapingure, Munyaradzi Paul...P05.02 Marathe, Jai G....P40.02, P4.9 Marco, Andrés...OA29.06 Marfil, Silvia...P6.0, P6.04 Margolis, David...OA03.02 LB Margolis, Leonid...OA30.06 LB Maric, Danijela...P25.02, P40.05 Mark, David...P23.6 Marlin, Romain...P40.20 Maroa, Jennifer...P4.23 Marovich, Mary... P26.02, P26.6, SY.04 Marrazzo, Jeanne... SY0.0, OA0.06 LB, P05.0, P36.03, P36.04, P42.03, P49.05, P49.06 Marrow, Matthew... P0.0, P27.06 Marshall, Dawn J....OA06.02 LB, OA2.06 LB Marshall, Kay...OA9.03 Martin, Darren P....P25.09 Martin, Jeffrey N.... P37.03, P43. Martin, Loïc...P35.04 Martin, Malcolm A....OA6.04, OA30.05, P0.2 Martin, Marsha A....P0.03 LB Martin, Romeo...P38.03 Martinelli, Elena...OA0.02 Martinez, Ana...P26.02 Martinez, Omar...P3.7 LB Martinez-Bonet, Marta... P5.04, P5.03 Martinez-Picado, Javier... P24.03, P26., P27.0 Martin-Gayo, Enrique...OA29.0 Martinon, Frederic...P27.02 Martinson, Francis...P4.0, P53.03 Martinson, Neil... P29.02, P29.03, P37.06 Martinson, Neil A....OA24.06 LB Maruta, Yasuhiro...P34.05 Marx, Preston...OA24.0 Marzinke, Mark...OA07.06 LB, OA3.0, OA3.02, OA3.05 LB, P8.06, P24.07 Masankwa, Geoffrey...P25.04 Mascola, John R....OA0.06, OA06.02 LB,OA06.04, OA06.06, OA2.0, OA30.05, PD05.02, P0.03, P0., P0.2, P5.25, P5.28 LB, SY0.0, PD03.03, P34.3 LB Maseko, Nombulelo...OA02.02, PD0.02 Masilo, Ntswaki R....OA09.02, P9.05, P23.04 Masiu, Alinah...P9.05 Mason, Rosemarie P34.3 LB Masopust, David...SY.02 Massinga-Loembé, Marguerite. P24.4 Masson, Lindi...OA04.03, OA2.02, P40.4, P40.9, P40.2 Maswai, Jonah...P23.02 Matanhire, Angeline...P23. Matano, Tetsuro... PD03.04 LB, P4.20 Matoba, Nobuyuki.... P8.04, P44.06 Matovu, Flavia K.... P02.04, P05.04, P23.0 Matsushita, Shuzo... P34.05, P35.03 Matthews, Lynn T....OA23.03 Matthews, Philippa...P24.9 Mattioli, Sandro P09.03 Mattoo, Hamid...OA25.02 Matubu, Allen T....OA20.05 LB Matus-Nicodemos, Rodrigo..OA30.05 Mauck, Christine...OA3.03, P23.5, P25.0 Maueia, Cremildo...P26.6 Mavrothalassitis, Orestes...P24.0 Max, Nibert...P4.06 Mayanja-Kizza, Harriet...P24.4 Mayer, Benjamin...P25.05 Mayer, Kenneth H.... OA07.04, OA07.06 LB, OA09.06, OA2.05, PD06.04 LB, P06.0, P08.03, P7.0, P7.02, P9.02, P43. Maynard, James P....P07.02 Mayo, Ashley...PD0.0, P02.0 Mayosi, Bongani...P5.5 Mayr, Luzia...P6.03, P34. LB Mazzotta, Francesco...OA08.02, PD02.04 Mbabazi, Elizabeth...P.04 Mbogua, Judie...P46.04 Mboup, Aminata...P3.8 LB Mboup, Souleymane...P3.8 LB Mbulawa, Zizipho...P40.07, P40.4 Mbunda, Theodora...P4.2 Mburu, Margaret...P48.04 Mc Cormack, Sheena...P26.08 Mc Gilvray, Marcus...P29.0 McBride, Ryan...OA25.04 McCaul, Nicholas...OA8.02 McConnell, Jason...OA3.02 Mccormack, Sheena...OA02.04, P40.2 McCorrister, Stuart J....PD02.02 McCoy, Clare... OA26.03, P4.03, P4.06, P4.07, P4.08, P5.4 McCoy, Connor... OA08.04 McCoy, Jay B....P26.5 McCutchan, Francine...OA2.06 LB Mcdermott, Adrian...OA6.0, OA25.0, P03.04 LB McElrath, Juliana...OA04.05 LB, OA.06 LB, OA6.06, OA7.06, P03.0, P24.06, P24.07, P24.08, P24.6 PD03.0 LB McEwen, Owen...P48.03, P48.05 McGlynn, Margaret...P47.07 AUTHOR INDEX 427

428 Author Index AUTHOR INDEX McGowan, Ian...SY07.0, OA03.06 LB, OA3.05 LB, OA27.06 LB, P23.03, P23.8, P33.03, P40.04, P44.03 McGuire, Andrew T....OA0.05 McGuire, Erin...P03.05 LB McIntyre, James A....RT0.0 McKay, Paul F.... P27.03, P4.3 McKee, Krisha...OA06.06, PD05.02, P5.25, PD03.03 Mckenna, Kevin...P46.0 McKinnon, Lyle R....OA0.04, OA7.03, P24.04, P24.05, P24.3, P40.9 McLellan, Jason S.... OA08.04 Mcmahan, Vanessa...P9.02 McMorrow, Martin...P26.04 McNevin, John...P24.6 McRaven, Michael...P25.02 Mdanda, Sanele P23.08 Mdluli, John...OA9.05 Meehan, Conor...P39.03 Meggi, Bindiya...P26.6 Mehendale, Sanjay M....P36.05 Mehrotra, Megha...P9.02 Meijer, Alexandra...OA07.03 Meiring, Tracey...P40.2 Meiser, Andrea...P5.0 Mejías-Pérez, Ernesto...P0.07, P4.05 Melbourne, Kathy...P7.0 Melendez, Johan...P23.5 Meli, Josephine...P34. LB Mellors, John W....OA27.0, P22.04 Mendoza, Janess...P0.0, P26.5 Mendoza, Mark G....OA08.03 Mengistu, Meron...OA25.03 Menis, Sergey... OA08.04, OA2.04 Menon, Mala D....P8.07 Mensch, Barbara...OA02.05, OA02.06 LB, OA5.02, OA5.03, OA5.04, P29.06 LB, P42.03, P42.04, P46.03 Menten, Joris...P40.5 Menu, Elisabeth...P2.0, P40.20 Mera Giler, Robertino...OA5.06 LB Merbah, Melanie...OA2.06 LB, P39.07 Mercuur, Clint...P5.3 Merlotti, Antonela...P24.02 Mermin, Jonathan...RT0.05 Mesa, Kathryn A....P0.0 Mesquita, Pedro...OA0.05, OA3.02, P8.03, P44. Mestecky, Jiri...OA24.0 Metch, Barbara...OA0.03, OA.04, P42.0 Mewalal, Nikoshia...OA29.02, P24.0 Meyer, Debra...P5.08, P5.02 Meyer, Laurence...P39.02 Meyn, Leslie OA3.0, P49.07, P53.04 Mgodi, Nyaradzo M....OA5.03, P23., P49.06, P53.03 Mhlanga, Felix...OA20.05 LB, P0.02, P53.03 Micci, Luca...OA2.03 Michael, Nelson...OA04.05 LB, OA05.04, OA08.04, OA.05, OA.06 LB, OA2.06 LB, OA4.03, OA2.06 LB, OA23.0, OA25.0, P03.04 LB, P23.02, P25., P26.02, P26.3, P26.4, P39.07, P44.09, SY.04 Micheni, Murugi.. P3.06, P3.6 LB Michiels, Jo...P44.05 Michiels, Johan...P35.0, P40.5 Michira, Peter...P43.2 Migueles, Stephen A OA6.0, P24.06 Milam, Douglas F....P29.04 LB Milanga, Maureen A....P43.2 Milford, Cecilia...OA23.04, OA26.0 Miller, Barbara...P42.04 Miller, Cari L....OA09.03 Miller, Christopher J....P03.04 LB Miller, Lori...OA5.0 Miller, Scott C....P33.06 Millier, Gérard...P09.4 LB Milovanovic, Minja... P29.02, P29.03 Mimiaga, Matthew J.... OA07.04, OA07.06 LB, PD06.04 LB, P08.03, P7.0, P7.02 Minja, Anna...P3.5 Miralles, Laia...P27.0 Miralles, Steve... PD0.04 Mirembe, Brenda G.... P05.0, P42.02, P42.05 Mishra, Anu...P36.04 Mishra, Anupam...P05.0 Missanga, Marco...P26.02, P26.08 Mitchell, Kate... OA27.04, OA28.05, P20.02 Mitchnick, Mark...OA3.02 Miti, Nokuthula...P53.02 Mizenina, Olga...OA03.05, P33.02 Mjösberg, Jenny...OA04.02 Mkhize, Baningi...P36.03 Mkhize, Nonhlanhla N....OA2.02, OA30.03, P3.03 Mkhize, Ntombifuthi...P23.06 Mlingo, Margaret...P23. Mlisana, Koleka...OA0.03, OA.04, P40.2, P42.0 Mncube, Zanele...OA29.03, P2.7 Mngadi, Kathryn T....OA9.06, P23.06 Mnqonywa, Nonzwakazi...P42.09 Mntambo, Adolphus...P40.09 Moate, Ronald...OA9.05 Mocello, A R....P43. Modeste, Mambo A....P09.06 Moelling, Karin...P4.28 Moench, Thomas...P40.0, P4.9 Mohanty, Soumyashree...P3.09 Mohns, Mariel...OA4.06 Mokaya, Tom...P45.06 Mokgoro, Mammekwa... P09.05, P48.02 Mokwena, Kebogile...P2.03 Moldoveanu, Zina...OA24.0 Moliki, Johnson M....OA20.02 LB, P05.09 LB Molinos, Luis M...P6.0 Molinos-Albert, Luis M....P6.04 Molloy, Peter...OA05.05 Moloelang, Maletsatsi...P38.03 Molyneux, Catherine...P3.06 Monaco, Daniela...OA2.0 Moncla, Bernard J....OA0.0, P49.07 Mondesire-Crump, Ijah...P24.20 LB Montague, Carl...OA9.06, P50.05 Montano, Silvia M....P24.08 Montefiori, David C....OA06.02 LB, OA.06 LB, OA2.06 LB, OA24.0, OA24.04, OA25.0, P03.04 LB, P0.0 Montgomery, Elizabeth T....OA02.0, OA02.05, OA02.06 LB, OA5.03, OA5.02, OA5.04, PD04.0, P4.0, P36.04, P42.04, P46.03, P50.04 Moodie, Zoe...OA0.03, OA.04, P42.0 Moodley, Amber...OA29.02 Moodley, Jayajothi...P05.07, P23.3, P32.0, P42.06 Moodley, Jeeva...P42.06 Moodley, Jothi...P45.05 Moodley, Nishila...P47.06 Moody, Michael A....OA06.02 LB, OA2.06 LB Moog, Christiane...P6.05, P6.08, P26.06, P33.03 Moore, John P....OA25.06 LB, OA0.03, OA25.04, OA25.05 Moore, Lizzie... OA23.04, OA26.0 Moore, Miranda S...OA6.06, P35.02 Moore, Penny L....OA06.05, OA2.0, OA2.02, OA30.03, PD05.02, P34.08 Moorhouse, Rika...P32.02 Moquin, Stephanie...P0. Morales, Javier F....P0.0 Morar, Neetha S....OA02.04, P07.05, P02.03, P07.06, P36.0 Moreau, Alain...P39.02 Moreno, Santiago...P5.03, P5.04 Morgan, Catharine...OA06.02 LB, P26. Morgan, Cecilia...OA7.06, P26.4 Morgan Jones, Molly...P9.03 Morgand, Marion...P39.02 Morin, Trevor J....P0.0 Morineau, Pascale...P26.0 Morioka, Hiroshi...P34.05 Morolo, M...P23.08 Morón-López, Sara...P26. Morris, Daryl...OA7.06 Morris, Lynn... PL0.0, OA06.05, OA.06 LB, OA2.0, OA2.02, OA30.03, PD05.02, P3.03, P34.08, P40.03, P4.26 Morrison, Charles...P25.0 Morrow, Kathleen M....PD04.02, P5., P5.2 Morton, Jennifer...OA28.0 Mosery, Nzwakie...OA23.02 Mosewu, Ireen...OA9.05 Moss, Bernard...OA.03, P26.02 Mothe, Beatriz...OA6.05, OA29.06, P24.03, P27.0 Motsamai, Petros J....P23.05 Mouquet, Hugo...OA0.05, P39.02 Moya, Eva...P3.7 LB Moyo, Thandeka...P34.03, P34.06 Mpanza, Lindiwe...P40.09 Mpendo, Juliet...P05.06, P23.07, P26.03, P26.07, P26.2 Mphili, Nonhlanhla...P30.06 Mpoke, Solomon...P33.0 Mubiana, Inambao...P05.08 Mubiru, Mike...P38.07 Muddiman, David C....OA26.06 LB Mudhune, Sandra...P23.08 Mueanpai, Famui...P25.08 Mueller, Kristen...OA20.0 Mueller, Monique...P.0 Mugisha, Emmanuel...PD0.05 Mugo, Nelly...OA27.02, OA28.0, OA28.02, P03.0, P24.07, P46.06, P52.06, PD02.0, P20.03, P46.05 Mugo, Peter M....P09.07, P3.06 Mugwaneza, Placidie... OA28.04 Mugwanya, Kenneth K....P20.03 Muhammad, Sevan...P25. Muigai, Anne...P45.06 Muik, Alexander...P4.2 Mujugira, Andrew...P52.06 Mukamuyango, Jeannine... P40.22, P49.08 Mukandavire, Zindoga...OA27.04 Mukhopadhya, Indrani... P3.0, P3.02 Mulder, Nicola...P3.03 Mulenga, Drosin M....P29.06 LB Mulenga, Joseph...P52.0 Mulenga, Muyunda...P09.0 Muller, Brian...P0.4 LB Müller, Christian...P24.8 Mullick, Charu...RT05.03 Mullick, Saiqa...P09.6 LB Mullins, James I....OA08.04, OA6.05, OA29.06 Mulondo, Jerry...P2.0, P36.0, P36.02 Munaiwa, Otillia...OA02.06 LB, OA5.03 Münch, Jan...P25.05, P25.07, P44.07 Munene, Elephas W....P25.06 Munir, Naeemah...P05.08, P42.0, P52.0 Muñoz, Eduardo...P5.03, P5.04 Muñoz-Fernández, Mª Ángeles..P5.0, P33.04, P33.05, P5.03, P5.04 Munseri, Patricia...P26.08 Muquingue, Humberto...P29.05 LB Muramatsu, Hiromi...P4.25 Murgor, Nereo...P49.0 Murillo, Wendy...P22.02 Murnane, Pamela M...P03.0, P24.07 Muro, Eva P....P22.03 Murokora, Daniel...PD HIV Research for Prevention 204 HIV R4P

429 Author Index Murphy, Diarmaid...OA03.04, OA26.03, P4.03, P4.06, P4.07, P4.08, P5.4 Murphy, Megan K PD03.02 Murugavel, K G....P34.0, P4.26 Musara, Petina...OA5.03, OA5.04, P4.0, P23. Musisi, Jane...P23.09 Musoke, Philippa...P38.07 Musyoki, Helgar...P07.08 Mutanha, Nyaradzo...P23.05 Mutengu, Lillian...P26.2 Muthumani, Kar...OA30.0 Mutisya, Elizabeth...P40.23 Mutisya, Leonard...P43.3 Mutonyi, Martin...OA28.03 Mutua, Gaudensia... P26.03, P26.04, P26.07, P26.2, P43.3 Mutuiri, Shem P.M....P25.06 Mutumwinka, Bella...P09.0 Muwawu, Rosemary...P23.09 Muzny, Donna...OA4.06 Muzoora, Conrad...P37.03 Mvandaba, Nomzamo...OA9.06, P23.06 Mviombo, P....P26.08 Mwaanga, Annie...P09.0, P09.05, P48.02 Mwamburi, Mkaya...P22.05 Mwangi, Irene...P26.04 Mwangi, Joseph...P49.09 Mwaniki, Lawrence...OA27.02 Mwanzo, Isaac...P30.04 Mwashigadi, Grace...P09.07 Mwatelah, Ruth S....P22.05 Mwathi, John N....P46.05 Mwau, Matilu...P45.06 Mwaura, Mary...P40.5 Mweemba, Oliver...OA02.04 Mwethera, Peter G....P25.06 Mwihaki, Lawrence M....P46.05 Mwimanzi, Philip M....P33.06 Myer, Landon...P50.0 Myles, Devin...OA24.0 Mylvaganam, Geetha...OA29.05 LB Mzimela, Misiwe A....OA02.04 Mzizi, Phumeza...OA9.05 N Nabel, G.B....OA06.02 LB Nabisere, Joselyne...P05.04 Nabukeera, Josephine...OA5.02 Naddunga, Faith...P49. Nagarajan, Karikalan...P09.08 Nagawa, Christine...P05.04 Naicker, Dshanta...P34.08 Naicker, Nivashnee... P40.03, P40.09, P40.7 Naidoo, Ishana...P38.03 Naidoo, Kewreshini...P2.7 Naidoo, Sarita...OA02.06 LB, P32.0, P42.06, P53.03 Nair, Gonasagrie...P0.02, P36.03, P36.04, P49.05, P49.06, P53.03 Nair, Lulu...P45.05 Nájera, José Luis...P4.05 Nakabiito, Clemensia...OA09.0, OA5.02, P0.02, P02.04, P05.0, P05.04, P23.09, P23.0, P36.03, P38.07, P42.02, P42.05, P49.05 Nakahara, Yusuke...P34.05 Nakato, Sarah...P05.05 Nakyanzi, Teopista...OA09.0, OA5.02, P02.04, P23.0, P42.02 Nalubega, Cissy L....P.04 Nalutaaya, Annet...P05.06, P23.07 Nambooze, Margaret...P.04 Namey, Emily... OA5.05, P.0, P46.0, P46.02 Nandudu, Norah...P42.07 Nangue, Achile Nangue...P2. Nangue, Achille...P2.4 Nanvubya, Annet...P05.06, P23.07 Nanyonga, Stella...OA09.0, P02.04, P23.0 Nanziri, Sophie C....OA09.0, P02.04, P23.0 Nardone, Anthony...OA07.03 Narpala, Sandeep...OA6.0 Nathalie Bosquet, Nathalie...OA08.06 LB Näveke, Arne...P47.07 Naw, Htee Khu...OA20.06 Nawaz, Fatima...OA7.02, P34.02 Nayager, Tashni...P38.04 Nazli, Aisha...OA20.0 Nchabeleng, Maphoshane...OA0.03, OA.04, P42.0, P50.0 Nchinda, Godwin...P2., P2.4, P4.22 Ndabambi, Nonkululeko...OA06.05 Ndadziyira, Pepukai...P23. Ndambuki, Rose...P40.23 Ndase, Patrick...PD0.0, P49.05 Ndawula, Patrick...OA09.0, P02.04, P23.0 Ndayisaba, Gilles...P40.5 Ndegwa, Jack...P43.3 Ndhlovu, Zaza M....OA20.04, OA29.02,P24.0 Ndibazza, Juliet...P36.0, P36.02 Ndlhovu, Zaza...OA29.03 Ndlovu, Bongiwe G....P4.23 Ndung u, Thumbi...OA04.02, OA4.04 LB, OA20.04, OA29.02, OA29.03, P09.05, P2.7, P24.0, P24.2, P34.2 LB, P40.08, P4.23, P48.02 Nduwamungu, Jean Nepo... P05.03, P3.02, P49.08 Ndwandwe, Duduzile... P4.09, P38.05 Ndzamela, Nwabisa...P38.03 Necochea, Edgar...P29.05 LB Nega, Melon T....OA7.05 Neikerk, Neliette V....P49. Nel, Annalene...OA09.02, OA3.06, P05.05, P09.8 LB, P5.09, P9.05, P2.02, P23.04, P49., P53.0, P53.02 Nelson, Julie A....OA22.06 LB, OA22.0, P03.05 LB Nelson, Wyatt...OA4.03 Neverov, Alexey D....P25.03 Neveu, Aurélie...P09.4 LB Newell, Marie-Louise...P52.05 Newling, Paul...OA7.06 Newman, Peter A....P.05 Newton, Robert...P49.02 Ng, Courtney...OA23.03 Ngai, Johnson... P6.03, P34. LB Nganou-Makamdop, Krystelle...OA4.0 Ngauy, Viseth...OA05.04, OA.05, P26.3, P44.09 Nga yo, Musa...P22.0 Ngcapu, Sinaye...P38.02, P40.2 Ngcobo, Nelisiwe...P50.05 Ngcobo, Nkosinathi...P23.08 Ngema, Isaac...OA9.05 Ng eno, Hillary...P36.08 Ngobi, Charles...P52.08 Ngongo, Bahati P....PD0.05 Nguay, Viseth...P26.02 Ngulube, Chepela...OA28.03 Ngure, Kenneth... OA27.02, OA28.0, OA28.02, P46.05, P46.06 Ngutu, Maria...P46.06 Nguyen, Richard... OA06.04 Nhambi, Leonel...P29.05 LB Nhamoyebonde, Sheperd...OA04.02 Nhlangulela, Lindiwe P38.03 Nicol, Melanie R....OA22.0 Niekerk, Neliette V....P05.05 Nielsen, Leslie...PD0.05, P05.06, P23.07 Nielsen, Susanne D....P2.07 Nieto, Jacobo...P4.05 Nilsson, Charlotta..OA.02, P26.08, P26.6 Nishimura, Yoshiaki...OA6.04, OA30.05, P0.2 Nitayaphan, Sorachai...OA04.05 LB, OA08.0, OA08.04, OA2.06 LB, OA4.03, OA2.06 LB, P25., P26.3, P26.4, P44.09 Njab, Jean... P09.2, P3.08, P49.04 Nji, Nadesh... P2., P2.4, P4.22 Njoku, Ogbonnaya...P23.02 Njoroge, Betty...OA02.0 Njuguna, Njambi...OA27.02 Njuguna, Stella...P48.04 Nkala, Busi...OA9.05, P50.0 Nkomonde, Nelisiwe...OA9.06 Nkompela, T...P23.08 Nkosi, Thandeka...OA29.02 Nkwihoreze, Hervette...P3.0 Noguchi, Lisa...OA09.0, P45.05, P45.07, P49.05, P50.04 Nogueira, Lilian...P6.03 Nomura, Takushi...P4.20 Nonhlanhla, Mkhize...P40.07 Nonodi, Thato P....P5.08 Noorbhai, Aslam...OA04.02 Novak, David S.... OA07.04, PD06.04 LB, P08.03 Novak, Rick...OA0.04 Nsanzimana, Sabin... OA28.04 Nsubuga, Rebecca N....P49.02 Ntjana, Hilda...P23.08 Ntombela, Fanele...P50.05 Ntombela, Fanelesibonge...OA9.06, P.02 Ntshele, Smanga...P9.05 Nugent, Sean...P5.9, P5.23 Nugeyre, Marie-Thérèse... P2.0, P40.20 Nunes, Rute...P5.22 Nunez-Curto, Aron...P43.03 Nunn, Kenetta...P40.6 Nussenzweig, Michel C.... SY05.0, P0., P6.03, P34.09, P39.02, P4.22 Nuttall, Jeremy...OA3.06, OA22.02, OA26.03, P4.0, P4.03, P4.06, P4.07, P4.08, P5.09, P8.05 Nuttens, Charles...P2.02, P2.08 Nyambi, Phillipe...P6.03, P34. LB Nyanga, Billy...P24.05, P40.9 Nyaradzo, Mgodi...P42.05 Nykoluk, Mikaela... P4.7, P4.8 Nyombayire, Julien... P05.03, P40.22, P3.02, PD03.04 LB, P49.08 Nzau, Benjamin...P33.0 Nzomo, Timothy J....P22.05, P25.04 O O Sullivan, Anne Marie...P25. O Connell, Robert J.... P26.4, P44.09, OA04.05 LB O Keefe, Barry...P33.02, P33.03 O Sullivan, Anne Marie...OA2.06 LB Oakland, Clare...OA07.03 Oates, Joanne...OA05.05 Obeng-Adjei, Nyamekye...P27.06 Obianwu, Otibho...P09.09, P49.04 Obregón, Patricia...P4.02 Ochieng, Washingtone... P22.05, P25.04 Ochoa, Maria T....P49.4 Ochola, Lucy A....P25.06 Ochsenbauer, Christina...PD03.0 LB O Connell, Robert J....OA05.04, OA08.0, OA08.04, OA.05, OA.06 LB, OA2.06 LB, OA2.06 LB, P25.08, P26.3 O Connor, David H....OA4.06 O Connor, Shelby L....P0.06 O Dell, Sijy...PD05.02, P5.25, P5.28 LB Odemijie, Godwin...P02.09 Odeyemi, Kofoworola...P3.3 Odhiambo, Jacob...OA5.05, P46.0, P46.02 Odoyo, Josephine...OA28.0, OA28.02 Odukoya, Olukemi...P3.3 Oehlmann, Wulf P26.06 Ofek, Adi...P5.2 Ofek, Gilad... OA06.06 Offerman, Kristy...P4.24 Ogilvie, Colin B....OA2.05 Oginni, Ayodeji...P09.09, P09.2, P49.04 AUTHOR INDEX 429

430 Author Index AUTHOR INDEX Ogola, Simon. P24.09, P26.04, P40.23 Ogolla, Irene...P45.08 Ogutu, Berhnards R.... P22.05, P25.04 Ogutu, Hilda...P40.23 O Hagan, Derek T....OA6.04 Oishi, Kevin...P43.5 LB Ojok, David...P38.07 Okala, Sandra G....OA24.05 O Keefe, Barry R....P33.0 Okesola, Nonhlanhla...P52.05 Oketch, Dismas C.O....P49.0 Okoboi, Stephen...P52.08 Okoko, Nicollate A....P45.08 Okolo, Felicia...P3.2, P30.05 Okonkwo, Chuks...P02.09 Okpokoro, Evaezi...P3.2, P30.05 Okumu, Eunice...OA02.03, P50.03 Olango, Kennedy O....P23.2 Olawo, Alice...P02.07, P2.04, P2.05, P48.06 Oldenburg, Catherine...OA07.04 Oldenburg, Catherine E....PD06.04 LB, P7.02 Olejniczak, Natalia...OA08.0, P2.06, P35.04, P44.04 Oliff, Monique...P07.04 Oloo, Florence A....P22.05 Olson, Gregory...OA20.04 Oluwasanu, Mojisola... P3.0, P45.0 Olvera, Alex... OA4.02, OA29.06 Omar, Anisa... P07.08, P09.07 Omar, Shatha S.A....P39.08 Ombati, Everlyn...P48.04 Omondi, Milton...P23.02 Omosa, Gloria...PD03.04 LB Omosa-Manyonyi, Gloria... P26.03, P26.04, P26.07, P26.2, P40.23 Ong wen, Patricia...P36.08 Onyango, Kevin O....P22.05 Onyango, Martin...P49. Operario, Don... P06.0, P07.08, P3.6 LB Oppert, Marydale... P3.0, P36.06, P49.08 Origoni, Massimo...P44.0 Orlandi, Chiara...OA30.02 Orlandini, Rodrigo P5.03 Orne-Gliemann, Joanna...P52.05 Oropeza, Celina...OA2.06 LB, P25. O Rourke, Sara M....P0.0 Osawe, Sophia...P3.2, P30.05 Osman, Nafissa...P26.6 Oster, Alexander F....SY.04 O Sullivan, Anne Marie...P39.07 Otibho, Obianwu...P09.2 Otsyula, Nekoye...P23.02 Ouattara, Gina... P26.04, PD03.04 LB, P40.23 Ouyang, Yong...OA05.0 Ouyang, Zhengyu...OA29.0 Ovejero, Hugo...P3.7 LB Overbaugh, Julie... OA7.03, P22.0, P26.9 Overman, Glenn...P03.05 LB Owino, George... OA9.02, P43.3, P45.08 Owiti, Patrick O....P36.08 Owuor, Kevin...P36.08, P45.08 Oyelakin, Oladayo T....P43.2 P Pabingui, Albertine...P09.4 LB Pace, Craig S....P39.02 Pack, Allison P.... P06.02, P06.03, P06.04, P2.04, P2.05, P23.4, P48.06 Padavattan, Nikita OA20.04, P40.08 Page-Shipp, Liesl...P26.03, P26.2 Pahar, Bapi...OA24.0 Paiardini, Mirko OA2.03 Palanee-Phillips, Thesla...OA02.02, OA02.04, OA09.04, PD0.0, PD0.02, P0.02, P02.0, P02.06, P36.03, P36.04, P38.08, P43.02, P45.05, P47.04, P49.03, P49.05, P49.06, P53.03, SY07.02 Palmer, Kenneth...P5.05 Palou, Elsa...P22.02 Palucka, Karolina...P24.0 Pan, Jason...P50.04 Pan, Ruimin...OA05.06 LB Pancera, Marie...OA0.06, P5.2, P5.28 LB, P39.02 Pancharoen, Kanokpan...P3. Panchia, R...P45.05 Panchia, Ravindre...P36.04, P49.05 Pandit, Hrishikesh...P40.8 Pannell, Jane...P40.28 Panousis, Constantinos...OA27.0 Pantaleo, Giuseppe...P0.07 Panther, Lori...OA09.06, P53.04 Paranjape, Ramesh...P09.08, P3.09, P36.05, P4.26, P48.0 Pardi, Norbert...P4.25 Parenzee, Penny...P43.4 Parikh, Urvi M....OA27.0, P22.04 Park, Chae Gyu...P4.22 Park, Harriet... PD03.04 LB, P26.04, P40.22, P40.23 Parker, R...P05.08 Parker, Rachel...OA28.04, P3.02, P3.0, P3.4, P36.06, P52.0 Parks, Christopher...PD03.04 LB Parks, Robert...OA06.02 LB, OA2.06 LB Parks, Thomas...OA8.04 Parniak, Michael...P5.06 Pascuccio, Massimiliano... OA7.02, P34.02 Passmore, Jo-Ann S.... PL03.02, OA04.03, OA7.0, OA2.02, P3.03, P40.03, P40.07, P40.4, P40.7, P40.9, P40.2 Patel, Mickey V....P40.24, P40.25 Patel, Sonal N....P34.09 Patel, Vainav...OA24.04, P4.0 Paterson, Pauline...P48.07 Patgaonkar, Mandar...OA8.0 Pathak, Reesab...SY.03 Pather, Arendevi... P0.02, P05.07, P53.03 Pathipvanich, Panita...OA23.0 Patil, Shilpa...OA0.04, P6.06, P34.0, P4.26 Pato, Sinazo...OA9.05, P07.04, P23.08 Pattacini, Laura...P03.0, P24.07 Pattanasin, Sarika... OA07.05, P09.3, P3., P36., P49.2 Patten, San...P48.03, P48.05 Patterson, Joey...P25. Patterson, Karen...P42.03 Patterson, Kristine B OA22.0, OA22.06 LB Patterson, Steve...P5.0 Patton, Dorothy L....OA8.06, P5.07, P5.20, P8.06, P8.08, P44.08 Pau, Chou-Pong...OA03.03, OA03.06 LB Paukovics, Geza...P35.02 Paul, Stéphane... P27.04, P4.03 Paula, Nersesian...P09.5 LB Paulson, James C....OA25.04 Pauthner, Matthias...OA0.03 Pavlakis, George N....OA6.05, OA24.04, OA29.06 Pavot, Vincent... P27.04, P4.03 Paximadis, Maria...P37.04, P37.06 Payne, Richard J...P35.02 Pedersen, Court...P2.07 Pegu, Poonam...OA04.04, OA06.03, OA25.0, P03.04 LB Peltekian, Cécile...P4.5 Peña, Ruth...P24. Peng, Binghao J....P26.7 Peng, Hong...P0.05, P4.6 Penney, Greg...P48.03, P48.05 Penrose, Kerri J....OA27.0, P22.04 Pensiero, Michael...OA05.02 Penton, Patricia K....P38.02 Perciani, Catia T....OA6.03 Perdiguero, Beatriz...P0.07, P4.05 Pereira, Lara...OA03.03 Pereyra, Florencia...OA2.05 Pereyra, Pehuen...P24.02 Perez, Erika...P22.02 Pérez-Alvarez, Susana...OA4.02 Perez-Brumer, Amaya...OA07.04 Perfetto, Stephen P.... OA06.04 Perkins, Benjamin... PD0.04 Perlmutter, Patrick...P40.0 Permar, Sallie...P03.05 LB Perouzel, Eric...P27.04, P4.03 Perrin, Helene...P2.02, P2.08 Perrot, Ludivine...OA03.04 Perry, Brian...OA02.03, P50.03 Perry, Jean...P40.28 Persaud, Deborah...RT02.03 Peters, Jennifer J....P5.9 Peterson, Bennett A....P24.06 Pfeifer, Nico...OA4.04 LB Phaahla, Manyabeane A P38.03 Pham, Ngoc...OA4.06 Pham, Phuc...OA2.06 LB Phan, Tran B....P03.04 LB, OA25.0 Phanuphak, Nittaya...OA05.04 Phanuphak, Praphan...OA05.04 Phelip, Capucine...P27.04 Phillip, Jessica L....P09.0, P53.03 Phogat, Sanjay...OA06.03, OA.06 LB, OA25.0, P03.04 LB Phonrat, Benjaluck...P44.09 Phuang-Ngern, Yuwadee...OA05.04 Phuti, Angel... OA7.0, P3.03 Pialoux, Gilles...P26.0 Pianowski, Luis...P5.03 Piatak, Michael...P4.08 Picker, Louis... SY09.0, SY.03,OA29.04 Pickett, Jim...OA9.03, RT0.03, PD0.04, P02.02, P9.03, P9.06 Picton, Anabela...OA04.03, P37.06 Piddubna, Anna I....P37.07 Pieralli, Stefano...P09.03 Pieterson, Ismelda...P09.5 LB Pilcher, Chris D....P25.07 Pillai, Shiv...OA25.02 Pillay, Deenan...SY03.03 Pillay, Diantha...OA02.03, P42.08, P50.03 Pillay, Yogan...RT03.04 Pillet, Stephane... P4.7, P4.8 Pillis, Devin M....P39.07 Pilon, Richard.OA6.03, P4.7, P4.8 Pineda, Juan A....P37.0 Pinter, Abraham...OA24.04, P34.07 Pion, Marjorie...P33.04 Piper, Jeanna...OA0.06 LB, OA3.05 LB, OA5.04, P42.03, P42.04, P42.05, P45.05, P45.07, P49.05, P49.06, P50.04 Piscitelli, Steve...OA03.02 LB Pise-Masison, Cynthia... OA04.04 Pissani, Franco...OA2.03 Pitcher, Annabell...P8.05 Pitisuttithum, Punnee...OA04.05 LB, OA08.0, OA08.04, OA.05, OA2.06 LB, OA4.03, P26.3, P26.4, P44.09 Pitoiset, Fabien...P27.05 Plagianos, Marlena. P5.27 LB, P49.03 Plana, Montserrat... P27.0, P4.03 Planelles, Lourdes...P4.05 Plebani, Anna Maria...P03.03 LB Plummer, Francis A....OA08.03, PD02.02, P03.02, PD02.05, P2.06, P24.04, P24.05, P24.3, P33.0, P37.02, P37.08, P4.7, P4.8, P40.3 Poignard, Pascal.. SY0.02, OA.0, OA2.02, P39.02 Poindexter, Alfred...OA3.03 Poizot-Martin, Isabelle...P26.0 Pokrovskaya, Anastasia V....P52.07 Pokrovsky, Vadim V....P52.07 Poli, Guido...P44.0 Politch, Joseph...OA24.02, P HIV Research for Prevention 204 HIV R4P

431 Author Index Pollakis, Georgios...P39.03 Radjabari, Sukaina...P49.08 Retornaz, Geneviève...P09.4 LB Rollenhagen, Christianne...P44. Pollara, Justin...OA2.06 LB, P03.05 LB Polonis, Victoria... OA23.0, P26.02, P26.08 Poltavee, Kultida...OA2.06 LB, P25., P39.07 Polyak, Christina...P23.02 Pool, Robert OA02.0, OA02.04 Poole, April...OA6.0 Poovan, Karmistha...P2.2 Poplimont, Hugo...P24.20 LB Porichis, Filippos...P24.0 Post Uiterweer, Arne...P47.0 Potenza, Nicoletta...P2.5 Pourtau, Pascal...P09.4 LB Powell, Eleanor A....P38.02 Power, Karen A....OA2.05 Powers, Kimberley A.... P0.0, SY03.0 Prado, Julia G....OA29.03, P24. Prego, Cecilia... P4.7, P4.8 Premrajh, Anamika...P0.02 Prentice, Heather...OA4.03 Preza, Gloria C....P49.4 Price, Mathew A.... Price, Matt...OA.0, OA4.0, OA2.0, OA2.03, P0.0, P09., P.04, P3.4, P36.0, P36.02, P49.08 Priddy, Frances...OA.0, P3.4, P26.03, P26.07, P26.2, P40.22, P40.23, P49.08 Primard, Charlotte...P27.04, P4.03 Prin, Pascale...P09.4 LB Prince, Heather M.A....OA22.0, OA22.06 LB, OA26.06 LB Prince, Jessica...OA.0, OA4.0, OA2.0, OA2.03 Prins, Henrieke...P09.07 Promda, Nutthawoot...OA07.05 Prudden, Holly J....P20.02 Pruvost, Alain...OA03.04 Psaros, Christina...OA23.02 Puangkaew, Jiraporn...OA.05 Pudney, Jeff...P40.02 Puertas, Maria C....P24.03, P26. Pugh, Daniel...P48.03, P48.05 Pulendran, Bali...SY.0, PD03.02 Pulerwitz, Julie...P3.8 LB Purcell, Damian F.J....P0.4 LB, P35.02 Q Qian, Han-Zhu...P29.04 LB Qin, Chuan...P4.6 Quaife, Matt OA27.03 Qualls, Zakiya...P34.07 Quatremère, Guillemette... P9.07 LB Quayle, Michael...P.03 Quillay, Heloise...P2.0, P40.20 Quinn, David S...OA25.0, P03.04 LB R Rabe, Lorna...OA0.06 LB, OA3.02, P53.04 Rahmati, Mona...P2.0, P40.20 Rainone, Veronica... PD02.04 Rainwater, Stephanie... OA7.03, P26.9 Rajaram, S...P20.02 Rakhmanina, Natella...P23.4 Ramafoko, Lebogang...RT0.02 Ramakant, Bobby...P02.05 Ramalo, Patience...PD0.02 Ramarathinam, Sri...P0.4 LB Rambally, Letitia...OA26.0 Ramchuran, Pranitha OA02.02, PD0.02, P38.08, P43.02 Ramesh, B M....P20.02 Ramirez, Kristel...P34.05 Ramjee, Gita...OA02.04, P0.02, P05.07, P09.0, P4.09, P23.0, P32.0, P36.03, P36.04, P36.0, P38.05, P38.06, P39.04, P42.06, P49.05 Ramlakhan, Yathisha...P34.2 LB Rammutla, Elizabeth...P38.03 Ramsey, Scott... PD06.0 Ramsland, Paul...P40.0 Ramsuran, Duran...OA04.02 Ranasinghe, Srinika...OA29.04, P24.0 Ranganathan, Uma Devi...P4.08 Rao, Mangala...OA25.0, P03.04 LB, P0.08, P26.8 Rao, Venigalla...P0.08 Rapiti, Ravikanthi...P09.6 LB Rastogi, Rachna...OA26.02 Ratner, Deena...P40.27 Ratto-Kim, Silvia..OA05.04, OA25.0, P03.04 LB, P26.02 Rautenbach, Clinton L....P.05 Raveendran, Muthuswamy...OA4.06 Rawat, Kavita...P44.0 Rawlings, Keith...OA5.06 LB Ray, Jocelyn...OA7.02, P34.02 Ray, Roslyn M OA20.02 LB, P05.09 LB, P33. LB Read, Sarah W....RT02.0 Reddy, Krishnaveni...OA02.02, OA09.04, OA8.0, PD0.02, P02.06, P33.07, P38.08, P43.02, P47.04 Reddy, Sharmila.. OA08.05, PD03.02 Reddy, Tarylee...P09.05, P48.02 Redon, Patrice...P09.4 LB Reed, Jason...SY.03 Rees, Helen...OA02.02, OA02.04, OA09.04, OA9.05, OA27.03, PD0.02, P02.06, P3.04, P23.08, P38.03, P38.08, P43.02, P46.04, P47.04, P50.0 Reeves, Iain...OA07.03 Reeves, Roger Keith...OA04.0, OA7.05, P2.09 Rehman, Khaja K....OA27.06 LB Ren, Li...P6.02 Rerknimitr, Rungsun...OA05.04 Rerks-Ngarm, Supachai...OA04.05 LB, OA08.0, OA08.04, OA.05, OA2.06 LB, OA4.03, P26.3, P26.4 Reushel, Emma...P27.06 Reyes-Terán, Gustavo...P22.02 Rhodes, Scott D....P3.7 LB Ribeiro, Susan...P4.04 Richards, Stephanie...P4.30 LB Richardson, Barbra...OA0.06 LB, P36.03, P36.04, P49.05 Richardson, Simone I....OA2.02 Richardson-Harman, Nicola...OA27.06 LB Richert-Spuhler, Laura E..... OA7.05, P2.05, P40.3 Richman, Douglas D....PD05.03 Richmond, Meika E OA08.03, P24.04, P24.05 Riddler, Sharon... P0.02, P23.8, P42.05 Rinehart, Alex...OA03.02 LB Ringe, Rajesh...P6.06 Rio Deiros, David...OA4.06 Rios, Daniel...OA29.05 LB Rittiroongrad, Surawach...OA.05 Rivero, Antonio...P37.0 Rizvi, Farrukh...P26.02 Roan, Nadia R....P25.05, P25.07 Roark, Ryan S....PD05.02 Robb, Merlin... SY.04, OA08.04, OA.02, OA.05, OA2.06 LB, OA2.06 LB, OA23.0, P23.02, P25., P26.02, P26.08, P26.4, P26.6, P39.07, P26.3, P44.09, OA04.05 LB Robbiani, Melissa...OA03.05, OA0.02, P4.02, P33.02 Robert, Wayne...P48.03, P48.05 Robert-Guroff, Marjorie...OA24.04, OA25.0, P03.04 LB Robins, Harlan...OA6.06 Robinson, Cortney...P09.0 Robinson, Hannah...PD05.03 Robinson, Harriet...OA05.03, OA08.05, OA.03 Robinson, James E....OA06.05, PD03.02, P34.07 Rocafort, Muntsa...OA29.06 Roche, Michael...P35.02 Rochereau, Nicolas...P27.04 Rodriguez, Ana Maria... PD05.04 Rodríguez, Aixa...P4.02, P33.02 Rodriguez de la Concepción, Maria L....P6.0, P6.04 Rodriguez-Garcia, Marta...OA20.03, P5.24 Roederer, Mario...OA06.04, OA25.0, P03.04 LB, P34.3 LB Roger, Thierry...P0.07 Rogers, Christine...OA9.04 Rogers, Jeffrey...OA4.06 Rogers, Paul M....OA24.05 Rohan, Lisa C....OA03.06 LB, OA3.0, OA8.06, OA22.02, OA22.03, PD04.02, P5.06, P5., P5.2, P5.20P5.05, P8.06, P8.08, P44.03, P44.08, P49.07 Rojas Castro, Daniela P.... P9.07 LB Rolland, Morgane...OA08.04, OA6.05, OA2.06 LB, P25. Roman, François...P26.07 Romano, Joseph...RT05.02 Romero, Lottie...P08.02 Ronald, Allan...P03.0, P20.03, P52.06 Rono, Kathleen...P25. Ronzière, Véronique...P09.4 LB Rosas, Miriam...P24.03 Rosati, Margherita...OA6.05, OA24.04, OA29.06 Rose, Annie...P26. Rosen, Elias...OA26.06 LB Rosen, Rochelle K....PD04.02, P5., P5.2 Rosenberg, Eric...OA2.05, P2.03 Rosenberger, Joshua G....P08.03 Rosenthal, Kenneth L.... P2.3, P40.26 Rosenthal, Matthew...P09.5 LB Ross, Greener...P30.06 Rossi, Lisa...P42.02 Rossoll, Richard M....P40.24, P40.25 Rouelle, Julie...OA06.05 Rouleau, Danielle... OA04.06 Rountree, Wesley P03.05 LB Rouskey, Craig...P4. Rousseau, Robert...P48.03, P48.05 Routy, Jean-Pierre... OA04.06 Roux, Surita OA.06 LB Ruan, Yuhua... P6.02, P29.04 LB, P39.0 Rubsamen, R M....P4. Ruffin, Nicolas...P24.0 Rugeles, Maria T....P04.02, P37.0, P2.5 Ruiz, Maria Julia...PD05.04, P24.02, P24.5 Russell, Marisa...P5.09 Russo, Aniello...P2.5 Russo, Julie...P44.03 Rutherford, Alexander R P39.0 Rutvisuttinunt, Wiriya...P25.08 Ruzagira, Eugene... P09., P.04, P2.0, P26.03, P26.07, P26.2, P36.0, P36.02 Rwanyonga, Richard...P09. Ryan, Elizabeth...P02.07, P2.04, P2.05, P48.06 S Saathoff, Elmar...P36.07 Saava, Margaret...P23.09 Saba, Elisa...P44.0 Sabatté, Juan...P24.02 Sabnis, Amit...OA04.05 LB Sacha, Jonah B....SY.03 Sachdev, Darpun D....PD06.03, P26.4 Sada, Ruth...P25.04 Safren, Steven A....OA07.06 LB, OA23.02, P08.03 Sagnia, Betrand...P2. Sahay, Seema...P36.05, P4.26 Saidi, Hela...OA08.06 LB AUTHOR INDEX 43

432 Author Index AUTHOR INDEX Sakala, Ephraim...OA28.03 Sake, Carol Stephanie N....P2. Sake Ngane, Carole Stéphanie.. P2.4 Salata, Robert...OA3.05 LB, P25.0 Salazar, Andres...P4.5, P4.22 Salazar, Ximena...P08.02, P43.03 Salazar-Gonzalez, Jesus...P24.7 Salgado, Maria...P26. Saliba, Regis...OA25.05 Salimi, Hamid...P35.02 Salomon, Horacio...PD05.04, P24.5 Samal, Sweety...P34.0 Samb, Badara... PD04.04 Sampathkumar, Raghavan...P37.08 Samsunder, Natasha... P40.03, P40.7 Samupindi, Shingirayi I....P23. Sanchez, Hugo...P52.03 Sanchez, Jorge...OA4.02, P24.08, P52.03 Sánchez, Cristina...P0.07 Sanchez-Palomino, Sonsoles..OA8.03 Sánchez-Rodriguez, Javier... P5.0, P33.05, P5.04 Sánchez-Sampedro, Lucas...P0.07 Sandberg, Johan K....OA7.04 Sanders, Eduard J....RT04.0, OA.0, P0.0, P09.07, P3.06, P07.08, P3.6 LB Sanders, Rogier W....OA0.03, OA8.02, OA25.05 Sanders-Buell, Eric...OA08.04, OA2.06 LB, P25., P39.07 Sandfort, Theo...P3.7 LB Sandhu, Manjinder...P49.02 Sandros, Marinella G....OA22.04 Sands, Anita...P09.04 Sandstrom, Paul...OA6.03, P4.7, P4.8 Sandström, Eric...OA.02, P26.6 P26.08, P4.2 Sang, Norton...P30.04 Santana, Vinicius...P4.04 Santoro-Rosa, Daniela...P4.04 Santra, Sampa...SY06.03 Sanyal, Anwesha...P40.27 Sanyu, Naomi...OA23.03 Saokhieo, Pongpun... PD0.04 Sardesai, Niranjan...OA6.05, OA24.0, OA24.04, OA29.06, OA30.0, P0.0, P26.02, P26.5, P27.06 Sariola, Salla...P3.06 Sarmento, Bruno...P5.22 Sarr, Moussa...P3.8 LB Sarwar, Shawn...P23.6 Satchwell, Karen...P0.04 Sather, D Noah...OA2.03 Sato, Alicia...OA.03 Sato, Hironori...P35.03 Sattentau, Quentin J....OA25.05 Satumay, Kesinee... OA07.05, P09.3, P3. Saukhat, Sergey R....P25.03 Saulle, Irma... OA08.02, PD02.04 Saunders, John...OA07.03 Savadsuk, Hathairat...OA.05 Sawe, Fredrick...P26.02 Saxena, Badri N....P02.05 Sayeed, Eddy...PD03.04 LB Saye-Francisco, Karen F....OA0.03, OA25.04 Scaglia, Fernando...P45.07 Scarlatti, Gabriella SY08.03, OA08.06 LB, OA.02, P03.03 LB, P26.6, P4.0 Schaefer, Malinda...OA4.04 LB, OA2.0 Scharf, Louise...SY05.0 Scheckter, Rachel...PD0.0, P02.0, P45.05 Scheepers, Cathrine...P34.08 Scheid, Johannes...OA0.05, P0. Scheider, Jeff... OA04.04 Schenkel, Jason...SY.02 Scherber, Sam...P52.0 Schief, William R....OA08.04, OA2.04, OA25.05, SY05.04 Schifanella, Luca...OA04.04, OA06.03, OA25.0, P03.04 LB Schiffner, Torben...OA25.05 Schiller, John T....PD03.05 LB Schlesinger, Sarah...P4.30 LB Schmid, Veronika P4.27 Schmidt, Claudia...P26.03, P26.07, P26.2 Schmidt, Stephen D....OA30.05, P0., P0.2 Schmidt, Sylvie...P6.05, P6.08 Schmitz, Frank...OA6.06 Schneider, Jeffrey R....P6.07 Schoen, Matthew K....OA24.02, PD05.03 Schomaker, Michael...P34.06 Schouten, Jeffrey...P07.05 Schrader, Thomas...P44.07 Schramm, Chaim...P34.08, PD05.02 Schreiber, Courtney A....OA3.03 Schuetz, Alexandra...OA05.04, OA08.0, P26.02 Schuler, Sidney...OA02.0 Schultheis, Katherine...P26.5 Schultz, Bruce T....SY.04 Schuster, Heiko...OA25.05 Schwartz, Jill L....OA0.05, OA3.03, OA3.04, P23.5, P45.07, P50.04, SY07.03 Schwartz, Katie...PD0.0, P02.0, P53.03 Schwartz, Olivier...SY2.03, P40.20 Schwing, Catherine... OA7.02, P34.02 Scott, Yanille... OA30.04 Scoulos-Hanson, Maritsa...P22.04 Scully, Eileen...OA4.0, OA2.03 Seaman, Michael...OA2.05, P6.03, P26.8 Seaton, Kelly...OA.03 Sebe, Modulakgotla...P50.0 Secor, Andrew...P3.6 LB Seddiki, Nabila...P24.0 Seder, Robert...OA6.04, P0.2 Seed, J...P05.08 Seemise, K...P23.08 Seese, Aaron...OA2.05 Seidel, Stephanie...P07.04 Seidor, Samantha...P4.02 Sekaly, Rafick...OA04.05 LB, OA25.0, P03.04 LB, P4.30 LB Sekiziyivu, Arthur...P25., P26.02 Sekoni, Adekemi...P3.3 Selepe, Pearl...P36.04, P45.05, P49.05 Semugoma, Paul...OA9.03 Sepúlveda-Crepo, Daniel... P5.0, P33.05 Serramia, María Jesús... P5.0, P33.05 Sette, Alessandro...OA29.04 Seyoum, Maaza...OA9.03, PD04.05 Shacklett, Barbara...P40.28 Shadabi, Elnaz...P37.08 Shade, Starley B...P36.08 Shaffer, Douglas N....PL04.0 Shahabi, Kamnoosh...P40.9 Shaheen, Nicholas J....OA22.06 LB Shahid, Aniqa...P33.06 Shanaube, Kwame...OA28.03 Shankar, Gita N....P5.04 Shanmugasundaram, Uma...P40.28 Shansab, Maryam...P6.07 Shao, Yiming...P0.05, P6.02, P29.04 LB, P39.0, P4.6 Shapiro, Lawrence...OA6.04, OA30.05, PD05.02, P0.2, P34.08 Shapiro, Roger...OA4.04 LB Sharkey, Tyronza... P3.0, P3.4, P36.06, P39. LB, P52.0 Sharma, Monisha...P09.7 LB Sharma, Shailendra K....OA0.02 Sharma, Siddharta...P24.8 Sharma, Sunita...OA03.06 LB Sharma, Vishnu L....P44.0 Shattock, Robin...OA08.0, OA08.06 LB, OA.02, OA7.0, OA22.05, OA24.05, OA30.06 LB, P2.06, P3.03, P4.0, P5.26 LB, P26.06, P27.03, P35.04, P40.06, P4.3, P44.04 Shaw, George...OA2.06 LB Shaw, Souradet Y....PD02.02 Shelke, Namdev...P5.6 Shell-Duncan, Bettina...P46.06 Shelter, Cory...P5.9 Shelus, Victoria...P06.02, P06.03, P06.04, P23.4 Shemshura, Andrey B....P25.03 Shen, Shaunna...OA2.06 LB, P6.07 Shen, Shen OA08.06 LB Shen, Xiaoying...OA06.03, OA24.04, OA25.0, P03.04 LB Shen, Zheng...OA20.03, P5.24 Sheng, Zizhang...OA6.04, OA30.05, P0.2 P34.08 Sherkar, Mahesh R....P8.07 Shetler, Cory...OA27.06 LB, P5.23, P8.03 Sheward, Daniel...OA2.02, P39.08 Sheward, Daniel J....OA06.05, OA2.0, P39.05 Shi, Wei...P5.25 Shipulin, German A....P25.03 Shisanya, Chris...P30.04 Shmelkov, Sergey...P26.8 Shorter, James...P44.07 Shrivastava, Tripti...OA0.04 Shroff, Ankit...P33.07 Shu, Tsugumine...PD03.04 LB Shukla, Brihaspati N....OA0.04, P6.06 Shukla, Praveen K....P44.0 Shultz, Andrew Z....P3.7 LB Shumba, Hildah...P48.02 Shuping, Liliwe...P39.06 Siangonya, Bella...P09.0 Sibanyoni, Jabhile M....P23.05 Sibanyoni, Maria...P46.04 Sibeko, Sengeziwe...OA30.03 Sibeko, Sylvia... OA02.02, PD0.02 Sibisi, Gugu...P23.08 Sibiya, Sibusiso...OA23.04 Sibiya, Sydney...P50.0 Siddiqui, Mohammad I....P44.0 Sidney, John...OA29.04 Siedner, Mark J....P43. Siegel, Aaron...OA27.06 LB Sievers, Stuart A....SY05.0, P34.09 Silbermann, Benjamin...P26.0 Silva-Santisteban, Alfonso.... P08.02, P43.03 Silveira, Paola...P5.03 Silvestri, Guido... OA2.03, PD03.02 Simbeza, Sandra...OA28.03 Simoni, Jane...P3.6 LB Simwinga, Musonda...OA28.03 Sinabamenye, Robertine...OA28.04, P05.03, P09.0, P3.02, P49.08 Sinangil, Faruk...OA2.06 LB Sinei, Samuel...OA2.06 LB Singh, Chitra...OA02.03, P50.03, P50.05 Singh, Devika...P49.05 Singh, Jasbir...P2.0 Singh, Jerome...OA9.06 Singh, Mahavir...P26.06 Singh, Manmohan...OA6.04 Singh, Sagri...P.03 Singh, Satya Prakash...P39.09 Singh, Udaya P....P33.08 Sinzinkayo, Denis...P30.0 Sips, Magdalena...OA24.03 Sironi, Manuela...OA08.02 Siskind, Rona...P07.05 Sithole, Kedibone...OA23.04 Sitoe, Nádia...P26.6 Siva, Samantha...P53.03 Siva, Samantha S....P23.3 Sivro, Aida...PD02.05 Skhosana, Joseph...OA5.05, P46.0, P46.02 Slack, Catherine M.... SY0.03, P07.03, P.03, P HIV Research for Prevention 204 HIV R4P

433 Author Index Smira, Ashley...OA06.05 Smit, Jennifer..... OA23.04, OA26.0 Smit, Jennifer A....OA23.02, P30.06 Smith, Adrian D....P07.08 Smith, Archer...P6.07 Smith, Davey M....PD05.03 Smith, Emilee...P50.0 Smith, James...OA03.03 Smith, Kieron A....P3.02 Smith, Kimberly...OA03.02 LB Smith, Stacey A....OA08.05, PD03.02 Smith-McCune, Karen...P40.28 Snead, Margaret C....P23.5 Snyder, Olivia...OA03.0, OA8.05 Sobieszczyk, Magda E....OA05.02, P07.02, P26.09, PD03.0 LB Socias, María E....P24.5 Socías, María Eugenia... PD05.04 Soderberg, K....OA06.02 LB Sodora, Donald L....PD03.02 Sodroski, Joseph...P5.28 LB Soghoian, Damien...OA29.04, SY.04 Sok, Devin... OA0.03, OA25.04 Solomon, Suniti...P34.0, P4.26 Solorzano, Norma...P22.02 Sonawane, Swapnil...P4.26 Songok, Elijah...P33.0 Soni, Sonal...P34. LB Sortijas, Steve... P50.03, P50.06 LB Sorzano, Carlos Oscar S....P0.07, P4.05 Sosa-Rubi, Sandra G....P06.0 Sosso, Samuel... P2., P2.4, P4.22 Soto, Cinque...OA06.06, P0. Soto-Torres, Lydia...PD0.0, P02.0, P53.03 Sougou, Amadou...P3.8 LB Soumaré, Marièma...P3.8 LB Southon, Lisa...OA07.03 Spadafino, Joseph...P3.7 LB Spagnuolo, Rae Ann...OA8.05 Spearman, Paul W....P26.9 Spencer, David...OA24.06 LB Spiegel, Hans M.L....P29.04 LB Spire, Bruno...P26.0 Spiro, Simon...OA8.02 Spreen, Bill...OA03.02 LB Srbinovski, Daniela...P40.29 Srinivasan, Sujatha...P40.3 Sriporn, Anuwat... OA07.05, P09.3, P49.2 Srithanaviboonchai, Kriengkrai...OA23.0, P09.02 Srivastava, Tripti...P34.0 Ssali, Livingstone...P52.08 Ssempiira, Julius...P05.06, P23.07 Ssetaala, Ali...P05.06, P23.07 Stablein, Donald M....OA06.03, OA25.0, P03.04 LB Stadler, Jonathan... SY0.04, PD04.0, P23.08, P46.04, P48.07 Stafova, Petra...OA04.05 LB, P4.30 LB Stamatatos, Leonidas... SY0.03, OA0.05, OA2.03 Stanton, Jill...P06.02, P06.03, P06.04, P23.4, P52.04 Starrett, Gabriel...OA4.06 Stein, Derek R....PD02.02 Steinbach, Jill M....OA26.04 Steinman, Ralph... P4.22, P4.30 LB Stevens, Marion L....P43.4 Stewart-Jones, Guillaume...OA0.06, OA25.05 Steytler, John...P53.02 Stieh, Daniel...P25.02, P40.05 Stockenstrom, Gail...P38.03 Stolarchuk, C....OA06.02 LB Stoner, Kevin A....OA20.05 LB Stout, Richard R....OA.02, P26.6 Stover, John...P47.07 Streeb, Gordon...P08.0 Streeck, Hendrik...SY.04, OA29.04 Street, Renee... P09.0, P23.0, P36.0, P42.09 Strode, Ann E....SY0.03 Strunk, Sarah...P05.03 Stuart, Andrew B....OA2.03 Su, Bin...P6.05, P6.08 Su, Hang...P24.20 LB Su, Ruey C....P37.02 Su, Ruey-Chyi...PD02.05, P03.02 Suarez, Carlos...P3.8 LB Sudi, L...P26.08 Sudyam, Ian T....P5.8 Sued, Omar...PD05.04, P24.5 Sugarman, Jeremy P.0 Sukhumvittaya, Suchada...OA05.04 Sukwicha, Wichuda... P3., P36., P49.2 Suleiman, Ibrahim... P09.2, P3.08, P49.04 Sun, Zehua...P6.09 Sunjeevan, Kershia...P50.05 Surenaud, Mathieu...P24.0 Suwa, Yoshiaki...P34.05 Suwanphatthana, Niwat...P07.07 Swann, Edith...OA.06 LB, OA7.06, PD03.0 LB, P24.08 Swanson, Michael...OA8.05 Swanstrom, Ronald...OA22.06 LB Swedrowska, Magda... P3.03, P44.2 LB Sweeney, Yvonne...OA8.06, P5.07, P8.06, P8.08, P44.08 Sweeton, Bentley...OA05.03 Sykes, Craig...OA3.03, OA3.04, OA22.06 LB, OA26.06 LB Sylla, Mohamed OA04.06 Syvertsen, Kristen...PD03.04 LB Szpara, Jordan J....P49.07 T Tabata, Nomzamo P....P38.03 Taccagni, Gianluca...P44.0 Taccarelli, Valerie...P09.03 Tachedjian, Gilda... P25.0, P40.0, P40.29 Takeda, Margaret...P40.28 Taljaard, Marthinette... P0.02, P36.03 Tanaka, Kazuki...P34.05 Tang, David... P4.7, P4.8 Tang, James...OA2.0 Tang, Jianming...OA.0, OA4.0, OA2.03 Tanser, Frank... SY03.02, SY03.03 Tanuri, Amilcar...P5.03 Tapia, Kenneth P22.0 Tarabureka, Noah...P29.02, P29.03 Tarimo, Edith...P4.2 Tartaglia, James...OA25.0, P03.04 LB Taruberekera, Noah P05.02 Tatarintsev, Alexander...P33.03 Tatoud, Roger...P26.08 Tatsuno, Gwen P....P0.0 Taylor, Andrew...OA03.06 LB Taylor, Doug...P49.03 Taylor, Jamilah... P06.02, P06.03, P06.04, P23.4 Tchadji, Jules C.... P2., P2.4, P4.22 Tecleab, Teghesti...OA.02 Tedesco, Jacquelynne...OA25.02 Tegha, Gerald...P03.05 LB Tekirya, Emmanuel...P24.2 Teleshova, Natalia...P4.02 Teller, Ryan...OA26.02 Tembe, Nelson...P26.6 Temgoua, Edith...P2., P2.4 Tempelman, Hugo...P53.02 Terahara, Kazutaka...P4.20 Terlikowski, Jessica... P9.03, P9.06 Terris-Prestholt, Fern... OA27.03, OA28.05, P20.02 Theis, James...P34.07 Theolis, Richard...P0.0 Thiebaut, Rodolphe... OA08.06 LB, P4.5 Thielemans, Kris...P27.0 Thienkrua, Warunee...P36. Thiong o, Alexander...P09.07 Thior, Ibou...P3.8 LB Thobakgale, Christina...OA29.03, P2.7, P40.08 Thomas, Katherine K.... P03.0, P24.07 Thomas, Kim...P48.03, P48.05 Thomas, Paul...P24.6 Thomas, Rasmi...OA04.05 LB, OA4.03, OA2.06 LB Thompson, Corbin...OA26.06 LB Thompson, Devon...P03.04 LB Thompson, Kerry...OA27.02 Thompson, Melanie...OA05.03 Thomson, Caspar...P43.04 Thomson, John...P3.0 Thurman, Andrea R....OA3.03, P23.5, P40.30, P44., OA0.05 Thurston, Sarah...P47.05 Tian, Sai...P0.09 Tichacek, Amanda...RT04.05, OA20.06, OA28.04, P05.03, P05.08, P08.0, P3.02, P3.0, P3.4, P36.06, P42.0, P49.08, P52.0 Tiemessen, Caroline T....OA04.03, OA24.06 LB, PD05.05, P37.04, P37.05, P37.06 Tietjen, Ian P33.06 Tindimwebwa, Edna...OA28.0, OA28.02 Tingey, Colleen...OA30.0 Tingstedt, Jeanette L P2.07 Tinsley, Jake...OA2.05 Tippanonth, Narongritt...OA07.05 Tiraby, Gérard...P27.04 Tjernlund, Annelie... OA7.04, PD02.03 Tkachenko, Elizabeth PD05.03 Tlagadi, Andrew...P38.03 Tober, Reinhard...P4.2 Todorova, Biliana...P27.02 Tolley, Elizabeth...P02.07, P06.02, P06.03, P06.04, P3.5, P2.04, P2.05, P23.4, P48.06 Tomaras, Georgia...OA05.02, OA06.02 LB, OA06.03, OA08.06 LB, OA.06 LB, OA2.06 LB, OA4.03, OA7.06, OA24.04, OA25.0, P03.04 LB, P03.05 LB, P6.07, SY06.0 Tomasicchio, Michele...P05.09 LB, P33. LB Tong, Tommy... OA06.04 Tongo, Marcel...P25.09 Tongtoyai, Jaray...P3., P49.2 Tooley, Len...P48.03, P48.05 Torjesen, Kristine...PD0.0, P02.0, P45.05, P45.07, P50.06 LB Torrieri-Dramard, Léa...P27.05 Totrov, Max...P0.3 LB Toulmin, Sushila A....P24.06 Tovanabutra, Sodsai...OA08.04, OA2.06 LB, OA23.0, P25., P39.07 Towers, Greg...P24. Trabattoni, Daria...OA08.02, PD02.04 Trakala, Marianna...P4.05 Trama, A.M....OA06.02 LB Tran, Alex P23.6 Tran, Karen...P0.04 Traoré, Ibrahima...P3.8 LB Travers, Simon...P5.3, P34.04 Travers, Simon A....PD05.0 Treijtel, Nicoline...P5.09 Tremblay, Cécile OA04.06 Tremblay, Michel J....OA20.0 Trenado, Emmanuel... P9.07 LB Trichavaroj, Rapee...OA05.04 Trinh, Hung V....P0.08 Tripathi, Raj K....P44.0 Trumpfheller, Christine...P4.30 LB Truong, Rosaline...OA0.02 Tsai, Alexander C....P43. Tshabangu, Nkeko C.... P29.02, P29.03 Tsibris, Athe...P4.06 AUTHOR INDEX 433

434 Author Index AUTHOR INDEX Tsitsi, Magure...P42.05 Tuff, Jeff...P37.08 Tuff, Jeffrey... P4.7, P4.8 Tully, Damien C OA2.05 Tulsi, Sahil...P40.03 Turgiev, Ali...P33.03 Turk, Gabriela...PD05.04, P24.5 Twefeho, Benjamin...P09. Tyler, Shaun... P4.7, P4.8 Tyssen, David...P40.0, P40.29 U Ugaonkar, Shweta...OA03.05 Ugen, Kenneth E....OA30.0 Ukpong, Morenike O.... P02.09, PD0.03, P07.0, P02.08 Ulrich, Michael...P32.03 Umaru, Stephen...P3.2, P30.05 Umviligihozo, Gisele...OA28.04, P3.02 Usmani, Shariq...P25.07 Uwamahoro, Honoree. P05.03, P3.02 V Vacas-Córdoba, Enrique...P33.04 Vaccari, Monica...OA04.04, OA06.03, OA25.0, P03.04 LB Valentin, Antonio...OA6.05, OA24.04, OA29.06 Vallabhjee, Leanne...P42.06 Vallely, Andrew...OA02.04 Valodia, Imraan...P50.06 LB Van de Stok, Mary...OA29.03 Van de Wijgert, Janneke...P40.5 Van der Elst, Elisabeth M....P07.08, P3.06, P3.6 LB Van der Horst, Charlie...P03.05 LB Van der Straten, Ariane...OA02.05, OA02.06 LB, OA5.02, OA5.03, OA5.04, PD0.0, PD04.0, P42.03, P42.04, P46.03, P49.05 Van der Veken, Pieter...P35.0 Van der Watt, Martin...OA09.03, P3.03 Van Gils, Marit J....OA0.03 Van Hateren, Andy...P26.05 Van Niekerk, Neliette...OA3.06, P5.09, P53.0, P53.02 Van Roey, Jens...OA03.04 Van Rompay, Koen...P4.08 Van Rooyen, Heidi...P09.7 LB Van Ryk, Donald...OA7.02, P34.02 Vanderford, Thomas... OA7.05, OA2.03 Vandergrift, Nathan...OA06.02 LB Vanham, Guido... P35.0, P39.03, P40.5, P44.05 Vanhooren, Leen...OA03.04 Varangrat, Anchalee... OA07.05, P30.02, P36. Varese, Augusto...P24.02 Vargas, Juan...P45.07 Vargas, Sara...P5. Vargas-Inchaustegui, Diego A....OA24.04, OA25.0, P03.04 LB Vasan, Sandhya...OA.05, OA2.06 LB, P26.3, P44.09 Vazquez, Thomas...P27.05 Veazey, Ronald S....OA08.0, OA22.05, P25.02, P6.07 Veldsman, Hanlie P23.04 Velhal, Shilpa...P4.0 Velu, Vijayakumar...OA29.05 LB Venables, Suzanne...P26.06 Venessa, Maseko...P40.07 Venkatraj, Muthusamy...P35.0 Venter, W.D. Francois... P3.04, P46.04 Venter, Willem D.F....P23.05 Ventura, Abigail B....SY.03 Venzon, David...OA06.03, OA24.04, OA25.0, P03.04 LB Vereecken, Katleen...P35.0 Verlinde, Carl...PD03.04 LB Vermund, Sten H....P29.04 LB Verrier, Bernard...P2.08, P27.04, P4.03 Viana, Raquel...P22.04 Vicenzi, Elisa...P44.0 Vickerman, Peter... OA27.03, OA27.04, OA28.05, P20.02 Viegas, Edna O....P26.6 Vigneron, James...P27.05 Viljanen, Miles...OA05.03 Villarreal, Daniel...P27.06 Villinger, Francois...PD03.02 Vincent, Heather...OA03.0 Voges, Maike...P4.28 Von Laer, Dorothee...P4.2 Vuidepot, Annelise OA05.05 Vuma, Amukelani...OA02.02, PD0.02 Vusha, Sophie...P46.06 Vuta, Christine...P23. Vwalika, Bellington...OA20.06, P05.08, P08.0, P42.0, P52.0 W Wachihi, Charles...OA08.03, P24.04, P24.05 Wagner, Andreas...P26.06 Wagner, Ralf... P0.07, P4.27 Wahl, Angela...OA8.05 Wahome, Elizabeth...P09.07 Wahren, Britta...OA.02, P26.08, P26.6 Wainberg, Mark P33.0 Wakefield, Steven...OA9.04 Walker, Bruce...OA04.02, OA2.05, OA20.04, OA29.0, OA29.02, OA29.03, OA29.04, P34.2 LB, P24.0, P40.08, P4.23 Wall, Kristin...OA20.06, OA28.04, P05.08, P08.0, P23.6, P52.0 Wallace, Melissa...P.05 Wallace, Zoe...OA05.05 Wallis, Carole L....P22.04 Walsh, Terri...P53.0 Walters, Jewel...P27.06 Wamalwa, Dalton...P22.0 Wambaya, Jeffrey W....P43.3 Wambuzi, Mathias...P05.06, P23.07 Wand, Handan... P4.09, P23.0, P36.0, P49.0 Wandera, Ronald...P45.08 Wang, Lin...OA22.02, P44.03 Wang, Shixia...P0.3 LB, P26.8 Wang, Shuhui...P0.05, P4.6 Wang, Ying-Ying...P40.6 Wangisi, Jonathan...P52.08 Ward, Andrew B....OA0.02, OA25.04, OA25.05 P0.04 Warrasally, Nabeela...P50.05 Warren, Emily...P48.07 Warren, Mitchell...PD06.02, P47.0, P47.02, P47.03, P47.07, RT03.05 Washington, Reynold. P20.02, P36.05 Washington Parks, Robyn... OA04.04 Wasinrapee, Punneeporn...P49.2 Wassenaar, Douglas R...P42.08 Watkins, Phaedrea...OA9.04, P52.04 Watnick, Dana L....P5.02, P23.7 Watson, Christopher C.... OA07.0, OA9.04 Watson, Sharon...OA02.03, P50.03, P50.06 LB Watts, Charlotte...OA27.03 Watts, D H....P45.07 Watts, Heather...P50.04 Wegmann, Frank...OA25.05 Wei, Danlan...OA7.02, P34.02 Wei, Huamian...P39.0 Wei, Qiang...P4.6 Weinberg, Aaron...P2.5 Weiner, David...OA24.0, OA30.0, P0.0, P26.02, P26.5, P27.06 Weiner, Debra...OA3.03 Weinfurter, Jason...OA4.06 Weinman, Renee...P23.8 Weis, Julie F....P26.9 Weiser, Sheri D....P43. Weiss, Helen...SY04.0 Weissman, Drew...P4.25 Wekesa, Clara...P49.02 Welsh, Sabrina... PD03.04 LB, P.03 Welte, Alex...P47.06 Wen, Yingxia...P0.09 Were, Edwin...P49.0 Werner, Lise...OA2.02, P3.05, P40.03, P40.09, P40.7, P40.2 West, Brian...P43.04 West, Jr., Anthony P....SY05.0 Westby, Mike...P35.02 Westmacott, Garrett...OA0.04, PD02.02 Wetzel, Katherine S....PD03.02 Whaley, Kevin...OA30.04, P34.0, P4.9 Wheatley, Adam...OA6.0 Wheeler, Darrell P....OA07.0 Wheeler, Lee Adam..OA4.05, P33.09 Wheeless, Angie...P23.5 Whipkey, Kimberly...P43.4 Whitacre, Ryan... PD06.03 White, David...OA07.03 White, Nicole...OA22.06 LB White, Rhonda...OA09.0, P02.04 Whitesides, John F....OA06.02 LB, OA2.06 LB Whitney, Stephen...OA04.04, OA25.0, P03.04 LB Wibmer, Constantinos Kurt...OA06.05, OA2.0, OA2.02 Wieczorek, Lindsay...OA23.0, P26.02 Wilcher, Rose...OA02.0 Wilkinson, Peter...OA04.05 LB Willems, Elisabeth...P39.03 Willer, David O....OA6.03 Williams, Constance...P0.3 LB Williams, Ifor...OA29.05 LB Williams, Katherine L....P26.9 Williams, Kathy...OA05.03 Williams, Marie...P26.06 Williams, Peter...OA03.0, OA27.06 LB Williams, W.B....OA06.02 LB Williamson, Anna-Lise...OA04.03, OA7.0, P3.03, P40.07, P40.4, P4.09, P4.24 Williamson, Carolyn...OA06.05, OA2.0, OA2.02, P39.05, P39.08, P40.2 Wilson, Douglas...P29.0 Wilson, Gregory J....OA05.02 Wilson, Ian A.... OA0.03, OA25.04 Wilson, Robert...P4.08 Wilton, James...P48.03, P48.05 Wimonsate, Wipas... OA07.05, PD0.04, P09.3, P3. Winaitham, Santi...P09.3 Winnall, Wendy R....P25.0 Wira, Charles R....OA20.03, P5.24, P40.24, P40.25 Wise, Megan...OA24.0, OA30.0, P0.0, P 26.5, P27.06 Wiseman, Roger...OA4.06 Witkowska, H. Ewa...P25.07 Wittenberg, Ralf W....P39.0 Woeber, Kubashni...OA02.06 LB, OA5.04 Wong, Fay E....P2.09 Wong, Gary... P4.7, P4.8 Wong, Patrick...OA30.05 Wood, Gemma...OA07.03 Wood, Natasha...P5.3, P34.04, PD05.0 Woodman, Zenda...P39.06, P39.08 Woodrow, Kim A....SY07.04, OA26.05, P5.8, P8.02 Woods, Robert J....PD05.0 Woodsong, Cynthia...P4.0 Worodria, William...P24.4 Wrin, Terri...OA2.02 Wu, Elwin...P3.7 LB Wu, Helen...SY.03 Wu, Kathleen... P05.08, P36.06, P42.0 Wu, Xilin...P4.29 Wu, Zhiwei...P HIV Research for Prevention 204 HIV R4P

435 Author Index Wyatt, Christina...P20.03 Wyatt, Richard...OA0.02, PD03.03, P0.04 X Xing, Hui...P39.0 Xu, Yongxian...OA05.03 Y Yacobson, Irina...P46.02 Yadon, Marisa...OA04.02 Yafant, Somsak...P09.3 Yahyaei, Sara... PD02.04 Yamamoto, Hidemi...P4.06 Yamamoto, Takuya...OA30.05 Yan, Celine...P26.06 Yan, Jian...OA24.0, P0.0, P27.06 Yandura, Sarah...OA22.02 Yang, Haitao...P5.05 Yang, Hongbing...OA05.05, P26. Yang, Jun...P34.02 Yang, Kuo H....OA3.03, OA22.06 LB Yang, Kuo-Hsiung...OA3.04 Yang, Yongping...OA0.0, OA0.06, P0. Yang, Zheng...OA6.02, P6.09 Yao, Xiao-Dan...P2.3, P40.26 Yin, Huazhong...OA6.02 Yin, Lu...P29.04 LB Yokoyama, Masaru...P35.03 Yola, Ntando... P07.03, P43.2 Yoon, In-Kyu...P25.08 Yoshimura, Kazuhisa...P35.03 Yotruean, Kriengkrai...P09.02 Younes, Naji P40.0, P40. Young, Liz H....P49.02 Young, Mary...P40.0 Yousefieh, Nazita...P40.30, P44. Yu, Bin...P0.0 Yu, Tianwei...OA4.0, OA2.03 Yu, Xiaoqiong...P03.02 Yu, Xu G....OA29.0 Yu, Xuesong...OA.03 Yu, Yi...P2.09 Yuan, Andrew... P5.7, P43.0 Yuan, Tingting...OA6.02 Yuan, Xin-Yang P4.7, P4.8 Yuan, Zhe... P4.7, P4.8 Yudin, Mark...OA7.03 Yurin, Oleg G....P52.07 Z Zachariah, Devika...PD03.04 LB Zakowicz, Anna...P43.04 Zalenskaya, Irina A.... P33.03, P40.30 Zapata, Wildeman... P04.02, P2.5, P37.0 Zeiser, Stefan...OA3.06 Zeitlin, Larry...P34.0 Zhang, Baoshan...OA06.06, P0., P5.25 Zhang, Jingyang...P36.03 Zhang, Meiyun...P6.09 Zhang, Mei-Yun...OA6.02 Zhang, Qicheng...P0.05 Zhang, Shimin...OA03.05, P5.27 LB Zhang, Yanyu...OA6.02 Zhang, Yu...P6.09 Zhang, Zhenghai...OA6.04, P0.2 Zhang, Zhou...P4.6 Zheng, Xin...P34.02 Zhou, Jingying...P4.4 Zhou, Tian...OA22.03, P44.08 Zhou, Tongqing... P0., P5.2, P5.28 LB Zhu, Whenhao...P8.05 Zhuang, Yan...P24.8 Ziegler, Susanne M....P2.6 Zirafi, Onofrio...P25.05 Zitzmann, Nicole...OA8.02 Zolla-Pazner, Susan...OA05.06 LB, OA4.03, P0.3 LB, P6.03 Zonca, Manuela...P4.05 Zulu, Michael Z....P2.7 Zurawski, Gérard...P4.5 Zurawski, Sandra...P4.5 Zydowsky, Thomas...OA03.05, P4.02, P33.02 Zydowsky, Tom...P5.27 LB AUTHOR INDEX 435

436 Notes 436 HIV Research for Prevention 204 HIV R4P

437 Notes 437

438 Notes 438 HIV Research for Prevention 204 HIV R4P

439 Tear out certificate along perforation. Certificate of Attendance HIV R4P 204 Conference This document is to certify that attended the HIV R4P 204 Conference held 28 3 October at the Cape Town International Convention Centre (CTICC), Cape Town, South Africa. Sharon Hillier University of Pittsburgh, United States Anatoli Kamali Medical Research Council/UVRI Uganda Research Unit on AIDS, Entebbe, Uganda Helen Rees Wits Reproductive Health and HIV Institute (Wits RHI), Johannesburg, South Africa Eric Hunter Emory University, United States Robin Shattock Imperial College, London, UK

440 Tear out certificate along perforation.

441 HIV R4P 204 Partners The HIV R4P conference would not be possible without the generous support of our conference partners. * + + ^ + The following institutions have contributed significant in-kind support: of this abstract book has been made possible through a financial contribution from the Government of Canada. The views expressed herein do not necessarily represent the views of the * Production Government of Canada. + HIV R4P 204 is made possible in part by the generous support of the American people through R3 AI from the National Institute of Allergy and Infectious Diseases (NIAID), and from the United States Agency for International Development (USAID). The views expressed in written conference materials or publications and by speakers and moderators do not necessarily reflect the official policies of the Department of Health and Human Services or USAID; nor does mention of trade names, commercial practices, or organizations imply endorsement by the U.S. Government. ^ The Joint United Nations Programme on HIV/AIDS (UNAIDS) is a sponsor of HIV R4P 204. The views expressed in conference materials or publications, by speakers and moderators, or by any conference sponsors do not necessarily reflect the official views or policies of UNAIDS; nor does mention of trade names, commercial practices, or organizations imply endorsement by UNAIDS.