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Quantitative analysis of anti–hepatitis C virus antibody–secreting B cells in patients with chronic hepatitis C

Authors

  • Takeji Umemura,

    1. Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD
    2. Department of Medicine, Shinshu University School of Medicine, Matsumoto, Japan
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  • Richard Y.-H. Wang,

    1. Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD
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  • Cathy Schechterly,

    1. Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD
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  • J. Wai-Kuo Shih,

    1. Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD
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  • Kendo Kiyosawa,

    1. Department of Medicine, Shinshu University School of Medicine, Matsumoto, Japan
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  • Harvey J. Alter M.D., M.A.C.P.

    Corresponding author
    1. Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, National Institutes of Health, Bethesda, MD
    • Department of Transfusion Medicine, Warren Grant Magnuson Clinical Center, Building 10, Room 1C711, National Institutes of Health, Bethesda, MD 20892-1184
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    • fax: 301-402-2965


  • Potential conflict of interest: Nothing to report.

Abstract

To investigate the quantitative characteristics of humoral immunity in patients with hepatitis C, we established an enzyme-linked immunosorbent spot (ELISpot) assay for detection of anti–hepatitis C virus (HCV)-secreting B cells. Receiver operating characteristic curve analysis demonstrated 100% specificity and 58% to 92% sensitivity for detecting B-cell responses to NS5b, NS3, E2, and core antigens. The median sum of anti-HCV–secreting B cells to all HCV antigens tested was significantly higher in 39 patients with chronic hepatitis C (47.3 spot forming cells [SFCs]/106 peripheral blood mononuclear cells [PBMCs]) than in 9 recovered subjects (15.3 SFCs/106 PBMCs; P = .05) or 11 uninfected controls (5.3 SFCs/106 PBMCs; P < .001); the significant difference (P = .018) in chronic versus recovered patients was in reactivity to nonstructural antigens NS3 and NS5b. Anti-HCV immunoglubulin M (IgM)–secreting B cells were also readily detected and persisted decades into HCV infection; there was no difference in IgM-positive cells between chronic and recovered patients. ELISpot reactivity to genotype 1–derived antigens was equivalent in patients of genotypes 1, 2, and 3. There was significant correlation between the numbers of anti-HCV IgG-secreting B cells and serum aminotransferase and to the level of circulating antibody. In conclusion, ELISpot assays can be adapted to study B-cell as well as T-cell responses to HCV. Measurement at the single-cell level suggests that humoral immunity plays a minor role in recovery from HCV infection and that B-cell immunity is strongest in those with persistent infection. (HEPATOLOGY 2005.)

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