When B Cells Go Bad: Infection, Inflammation and Chronic B Cell Stimulation
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1 When B Cells Go Bad: Infection, Inflammation and Chronic B Cell Stimulation Karen S. Anderson MD PhD Associate Professor, Biodesign Institute Arizona State University Mayo Clinic Arizona
2 Conflicts of Interest I serve on the advisory board and have received consultant fees and stock options with Provista Dx. No off-label clinical diagnostics or therapeutics will be discussed.
3 W. Michael Kuehl & P. Leif Bergsagel, 2002
4 Monoclonal Gammopathy of Undetermined Signficance 3.2% of adults over the age of 50; 6.6% of adults over age 80 Premalignant disease that can transition to myeloma; average 2-15 years, rate 1% per year Over 90% of patients with MM have a premalignant plasma cell disorder 50% Ig translocations; 50% hyperdiploid Errors in switch recombination and somatic hypermutation of the B cell Likely due to an abnormal response to antigenic stimulation
5 Chronic Infections and Antibodies EBV, HHV-8, HCV, (CMV), H. pylori are associated with B cell lymphoma and B cell chronic lymphocytic leukemia what about myeloma? Ig in HCV-positive myeloma patients can target the virus ~20% of MGUS and myeloma Abs target infectious antigens (ASH 2013) Would treatment of chronic infections also prevent or treat MGUS?
6 Chronic Infections not Associated with MGUS Bida and Rajkumar Mayo Clin Proc 2009
7 How B cells Proliferate in response to infection Moir and Fauci, Nat Rev Immunol 2009
8 Antibody structure: Ligands (antigens) What is the specificity of antibodies? What is the diversity of antibodies?
9 When Ig genes rearrange, there are >10 11 potential molecules
10 To facilitate research, DNASU stores over 162,000 plasmids and distributes these to researchers in 37 states and 38 countries
11 Nucleic Acid-Programmable Protein Array (NAPPA) Proteomics: the study of all of the proteins in the human body We use plasmids to make over 10,000 human proteins By putting these plasmids on a glass slide (NAPPA) we can make and study over 10,000 proteins at one time Protein Expression 1. Print Plasmids 2. Express & capture proteins Replicate arrays of candidate proteins Find antibodies in patients blood
12 NAPPA Array Production and Screening Gene Cloning Bacterial plating DNA preparation Array Printing
13 Antigen array Add patient serum p53
14 Serologic Biomarkers for HPV+ Cancer Human papilloma virus (HPV16): ~70% of oropharyngeal cancers Emerging epidemic in US and Europe HPV Genome: 8 ORFs Diagnosis Proportion surviving Number at risk NE2 positive NE2 negative Prognosis NE2, p<0.001 Kaplan-Meier survival estimates NE2 positive NE2 negative Follow-up time (months)
15 Identification of the Targets of Antibodies Protein microarrays are now used to test >10,000 proteins for antibody targets Large gene collections can be leveraged for rapid protein display We are developing pathogen-specific arrays to identify antibodies in blood We need to measure the antibody IMMUNOME to understand the pathogenesis of MGUS
16 What is the diversity of Antibodies? Can we detect specific Ig rearrangements?
17 DNA Origami nanostructure design Hao Yan and Joe Blattman, Biodesign
18 Overall strategy to obtain linked RNA sequences from single cells.
19 Conclusions Antibodies have extraordinary sequence diversity There are emerging technologies for quantitating that diversity By linking single-cell RNA capture with next-gen sequencing, we may be able to: Identify early events (?pre-mgus) of loss of diversity Rapidly generate patient-specific probes for molecular detection of rearrangements
20 Acknowledgements Anderson Lab Julia Cheng Ting Li Rizwan Alam Benjamin Katchman Krishna Sundaresan Diego Chowell Shay Ferdosi Hans Frykman I.Purushothaman ASU Biodesign Institute Joseph Blattman Hao Yan Josh LaBaer Ji Qiu Garrick Wallstrom Laura Gonzalez Jin Park Fernanda Festa Fernando Hernandez Mayo Oncology Robert Brown Don Northfelt Alison Goulder Doug Lake Jack Resnik Barb Pockaj Peter Chang Michael Barrett Our Patients NCI/Early Detection Research Network Zicarelli Foundation
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