1. Direct Ex Vivo Functional Analysis of HCV-Specific T Cells.
- Author
-
Park SH and Shin EC
- Subjects
- Antigens, Viral immunology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Cell Culture Techniques instrumentation, Cell Culture Techniques methods, Cell Proliferation, Cells, Cultured, Cytokines immunology, Drug Development methods, Enzyme-Linked Immunospot Assay instrumentation, Enzyme-Linked Immunospot Assay methods, Flow Cytometry instrumentation, Flow Cytometry methods, Hepatitis C blood, Hepatitis C immunology, Hepatitis C virology, Humans, Immunogenicity, Vaccine, Leukocytes, Mononuclear, Lymphocyte Activation immunology, Viral Hepatitis Vaccines immunology, Cytokines analysis, Hepacivirus immunology, Hepatitis C prevention & control
- Abstract
CD8
+ and CD4+ T cells play a critical role in viral clearance during self-limited HCV infection. Moreover, induction of robust and polyfunctional memory T cells is a primary goal in the development of T-cell-based HCV prophylactic vaccines. Direct ex vivo function of HCV-specific T cells without in vitro expansion can be assessed by measuring cytokine secretion or cell proliferation upon in vitro stimulation with HCV antigens using enzyme-linked immunospot (ELISpot) assays, intracellular cytokine staining (ICS) assays, and flow cytometry-based T-cell proliferation assays. In particular, polyfunctionality of T cells can be assessed by ICS assays using multicolor flow cytometry. Here, we provide protocols to measure direct ex vivo functions of HCV-specific T cells.- Published
- 2019
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