Clinical significance of routine immunological markers in patients infected with influenza type A H1N1 virus

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Susanna Wilson
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1 ,,,, ( ), : 2009 ( ) ( H1N1) 32 H1N1 T,,, H1N1 CD3 + CD3 + CD4 + CD3 + CD8 + T ( cells/ l), C3 C4( g/l) C ( mg/l),,, H1N1 : H1N1; T ; Clinical significance of routine immunological markers in patients infected with influenza type A H1N1 virus SHEN Fang, JIN Xin, MAO Hui-Jun, GAO Qiong, CHEN Dai-Hong Shanghai Public Health Clinical Center Affiliated to Fudan University, Shanghai , China Abstract: The paper aimed to investigate the differences in immunological evaluations between influenza type A H1N1 virus-infected patients and uninfected individuals. T-cell subtypes in peripheral blood from32 H1N1-infected patients were quantified using flow cytometry. Specific protein markers in serum were determined and blood cells were counted. The results showed that mean counts of CD3 +, CD3 + CD4 +, and CD3 + CD8 + T cells in peripheral blood of H1N1-infected patients were significantly reduced compared with those in uninfected individuals whereas serum complement components ( C3 and C4) and C-reaction protein were significantly increased in H1N1-infected patients. Furthermore, platelets, leukocytes, lymphocytes, and eosinophilic cells were significantly decreased, while monocytes were significantly increased in this population. It is indicated that routine immunological evaluation has clinical significance in H1N1-infected patients. Key words: Influenza type A H1N1 virus; T cells; Emerging and reemerging infectious disease ( ) ( H1N1),, ( ),,, H1N1, T : ( 2008 WSZB007) :, shenfang@ shaphc. org Corresponding author: SHEN Fang, shenfang@ shaphc. org :32 H1N :,, ( reverse transcriptase-polymerase chain reaction, RT-PCR) 20, 12 ; 21 52, , 75 mg 2, 5 d : 30, H1N1 15, 15 ; 24 55, CD3- FITC/CD8-PE/CD45-PerCP/CD4-APC, CD4-FITC CD8-FITC CD25-PE CD45RA-

2 Journal of Microbes and Infection, December 2009, Vol. 4, No PE CD45RO-PE CD3-PerCP,, BD BD FACSCalibur, Immage ACT5diff T 20 l CD3-FITC/ CD8- PE/CD45-PerCP/CD4-APC,, EDTA( 50 l/ ), 20 min; ( 1 10) 450 l/,, 10 min; Multiset CD4 + CD8 + CD45RA + CD45RO + CD4 + CD25 + 6, 10 l : 1, ; 2 CD4-FITC CD45RA-PE; 3 CD4-FITC CD45RO-PE; 4 CD8-FITC CD45RA-PE; 5 CD8- FITC CD45RO-PE; 6 CD4-FITC CD25- PE 100 l,, 20 min; 1 ml/, 10 min; 190 g 5 min, ; ( phosphate-buffered saline, PBS), ; 400 l PBS ; Cellquest , Immage IgG IgA IgM, C3 C4, ( transferrin, Trf) C ( C-reactive protein, CRP), ACT5diff 1. 4 mean SD, SPSS11. 0 P < H1N1 T CD3 + CD3 + CD4 + CD3 + CD % vs % % vs % % vs %, P > 0. 05, CD3 + CD4 + /CD3 + CD , P =0. 091, CD3 + CD3 + CD4 + CD3 + CD8 + ( cells/ l) vs vs vs , P ,, H1N1 CD3 + CD3 + CD4 + CD3 + CD8 +, ( 1) CD4 + CD25 + ( ) %, ( ) %, P = , ( ) CD4 + CD45RA + CD4 + CD45RO % vs % % vs %, P > 0. 05, CD8 + CD45RA + CD8 + CD45RO % vs % % vs 9. 98%, P > 0. 05, CD4 + CD45RA + /CD4 + CD45RO + CD8 + CD45RA + /CD8 + CD45RO + ( 2) 1 H1N1 T Tab 1. Comparison of T-cell subtype in peripheral blood of H1N1-infected patients and healthy individuals T-cell subtype H1N1( n = 32) Normal control ( n = 30) P value CD3 + ( % ) CD3 + ( cells / l) * CD3 + CD4 + ( % ) CD3 + CD4 + ( cells / l) * CD3 + CD8 + ( % ) CD3 + CD8 + ( cells / l) * CD3 + CD4 + /CD3 + CD * P <

3 200 2 H1N1 CD4 + CD45RA + CD4 + CD45RO + CD8 + CD45RA + CD8 + CD45RO + Tab 2. Comparison of CD4 + CD45RA +, CD4 + CD45RO +, CD8 + CD45RA + and CD8 + CD45RO + in peripheral blood of H1N1-infected patients and healthy individuals T-cell subtype H1N1( n = 32) Normal control ( n = 30) P value CD4 + CD45RA + ( % ) CD4 + CD45RO + ( % ) CD4 + CD45RA + /CD4 + CD45 RO CD8 + CD45RA + ( % ) CD8 + CD45RO + ( % ) CD8 + CD45RA + /CD8 + CD45 RO H1N1 C3 C4 CRP Trf IgG IgA IgM ( g/l) vs vs vs 1. 48, P > 0. 05, C3 C4 ( g/l) CRP ( mg/l) vs vs vs 2. 68, P <0. 05, Trf ( g/ L) ( vs 2. 50, P = ), H1N1 IgG IgA IgM, C3 C4 CRP, Trf ( 3) 2. 3 H1N1 ( 10 9 /L) vs 4. 47, P = , ; ( 10 9 /L) vs , ( 10 9 / L) vs 5. 98, ( P <0. 05),, ( 10 9 /L) vs % vs 0. 03%, ( P > 0. 05) % vs % ( P = ) vs 2. 09( P = ) ( 10 9 /L) vs vs 0. 12, P ,, ( % vs %, P = ), ( 10 9 /L) ( vs 3. 05, P = ) ( 10 9 /L), ( % vs 6. 95% vs 0. 42, P < 0. 05), H1N1 ( 4) 3 H1N1 Tab 3. Comparison of specific protein markers in serum in H1N1-infected patients and healthy individuals Serum protein H1N1( n = 32 ) Normal control ( n = 30) P value IgG ( g/l) IgA ( g/l) IgM ( g /L) C3 ( g/ L) * C4 ( g/ L) * Trf ( g /L) * CRP ( mg/ L) * * P < 0. 05; Trf: transferrin; CRP: C-reactive protein.

4 Journal of Microbes and Infection, December 2009, Vol. 4, No H1N1 Tab 4. Comparison of routine blood test results in H1N1-infected patients and healthy individuals Blood cells H1N1( n = 32 ) Normal control ( n = 30) P value PLT ( 10 9 /L) * WBC( 10 9 / L) * RBC ( 10 9 /L) NEUT ( % ) * NEUT ( 10 9 / L) LYMPH ( % ) * LYMPH ( 10 9 /L) * MONO ( % ) * MONO ( 10 9 /L) * EO ( % ) * EO ( 10 9 /L) * BASO ( % ) BASO ( 10 9 /L) * P < PLT: platelet; WBC: white blood cell; RBC: red blood cell; NEUT: neutrophil granulocyte; LYMPH: lymphocyte; MONO: monocyte; EO: eosinophil granulocyte; BASO: basophilic granulocyte. 3, 1918 ( H1N1) 2 000, : ( hemagglutinin, HA) ( neuraminidase, NA) ; [ 1] H1N1,, 3 [ 2, 3 ],, CD4 + T T ( CD8 + ) [ 4 ], 32 H1N1 T ( CD3 + ) T ( CD3 + CD4 + ) T / ( CD3 + CD8 + ) 30,,,, Perez-Padilla 61% H1N1 [ 5 ], [ 6], T ( CD4 + CD25 + ) T, ( World Health Organization, WHO),, H1N1, CD4 + CD25 + H1N1 CD4 + CD45 2 : CD45RA + CD45RO + CD45RA + T, CD45RO + T, T T, CD45RO + CD45RA + ;, T, T [ 7 ], CD45RA + CD45RO + [ 8, 9 ],, CD4 + CD45RA + CD4 + CD45RO + CD8 + CD45RA + CD8 + CD45RO + ; CD45RA + /CD45RO +, H1N1 CD45RA + CD45RO +,, CD45RO + /CD45RA +

5 202 IgG IgA IgM, H1N1 IgG IgA IgM, CRP Trf C3 C4,, C3 C4 CRP, CRP, Trf, [ 10 ],,, [ 11],,,,, [ 12, 13 ], H1N1 [ 14],, H1N1 CD3 + CD3 + CD4 + CD3 + CD8 + ; CD45RA + CD45RO + CD4 + CD25 + ; C3 C4 CRP ; ;, H1N1,, (, ), H1N1,, [ 1] Peiris JS, Poon LM, Guan Y. Emergence of a novel swine-origin influenza A virus ( S-OIV) H1N1 virus in humans[ J]. J Clin Virol, 2009, 45: [ 2] Shanta MZ, Donald SB. Historical perspective emergence of influenza A ( H1N1) viruses[ J]. NEngl J Med, 2009, 361: [ 3] Chowell G, Bertozzi SM, Colchero MA, et al. Severe respiratory disease concurrent with the circulation of H1N1 influenza[ J/ OL]. http: / / content. nejm. org/cgi/ reprint/nejmoa pdf [ 4],. [ M]. 2. :,2005: [ 5] Perez-Padilla R, de la Rosa-Zamboni D, Ponce de Leon S, et al. Pneumonia and respiratory failure from swineorigin influenza A ( H1N1) in Mexico[ J/ ON]. http: / / content. nejm. org/cgi/reprint/nejmoa pdf [ 6]. [ M]. : S. H. E., A., R... :, 2005:334 [ 7]. [ M]. : S. H. E., A., R... :, 2005: 189 [ 8] Ullum H, Cozzi L A, Victor J, et al. Increased losses of CD4 + CD45RA + cells in late stages of HIV infection is related to increased risk of death: evidence from a cohort of 347 HIV-infected individuals [ J]. AIDS, 1997, 11: [ 9] Maldonado A, Mueller YM, Thomas P, et al. Decreased effector memory CD45RA + CD62L - CD8 + T cells and increased central memory CD45RA - CD62L + CD8 + T cells in peripheral blood of rheumatoid arthritis patients [ J]. Arthritis Res Ther, 2003, 5 ( 2) : [ 10],,. [ M]. 3, :,2006, [ 11]. [ M]. :, 2009: [ 12],,,. A( H5N1) [ J]., 2006: 24( 4) : [ 13],,,. B19 [ J]., 2006, 21( 15) : [ 14] Franklin DZ, Pancake BA, Brown WH. Prevalence of HTLV-I Tax in a subset of patients with rheumatoid arthritis[ J]. Clin Exp Rheumatol, 2002, 20( 2) : ( : )

Collect and label sample according to standard protocols. Gently invert tube 8-10 times immediately after draw. DO NOT SHAKE. Do not centrifuge.

Collect and label sample according to standard protocols. Gently invert tube 8-10 times immediately after draw. DO NOT SHAKE. Do not centrifuge. Complete Blood Count CPT Code: CBC with Differential: 85025 CBC without Differential: 85027 Order Code: CBC with Differential: C915 Includes: White blood cell, Red blood cell, Hematocrit, Hemoglobin, MCV,