Flow-cytometric detection of vaccinia-induced memory effector CD4(+), CD8(+), and gamma delta TCR(+) T cells capable of antigen-specific expansion and effector functions
| Autor: | Joy Eslick, Robert B. Belshe, Thomas P. Monath, Getahun Abate, Frances K. Newman, Sharon E. Frey, Daniel F. Hoft |
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| Rok vydání: | 2004 |
| Předmět: |
CD4-Positive T-Lymphocytes
T cell Succinimides Vaccinia virus CD8-Positive T-Lymphocytes chemistry.chemical_compound Antigen Interferon medicine Vaccinia Immunology and Allergy Humans biology Carboxyfluorescein succinimidyl ester Receptors Antigen T-Cell gamma-delta T lymphocyte Flow Cytometry Fluoresceins Virology Molecular biology Infectious Diseases medicine.anatomical_structure Granzyme chemistry biology.protein Granzyme A Immunologic Memory CD8 medicine.drug |
| Zdroj: | The Journal of infectious diseases. 192(8) |
| ISSN: | 0022-1899 |
| Popis: | We developed a carboxyfluorescein succinimidyl ester (CFSE)-based flow-cytometric assay that can detect different subsets of vaccinia-specific T cells capable of both antigen-specific expansion and protective effector functions. Proliferation and effector functions were detected by CFSE dilution and intracellular staining, respectively. Absolute numbers of CD4(+)/CFSE(lo)/interferon (IFN)- gamma (+), CD8(+)/CFSE(lo)/IFN- gamma (+), CD8(+)/CFSE(lo)/granzyme A(+), and CD8(+)/CFSE(lo)/CD107a(+) T cells present after in vitro stimulation with live vaccinia were significantly higher in immunized individuals (P.05). These CD4(+) and CD8(+) T cell increases were2 log higher than increases detectable by standard lymphoproliferation and cytotoxicity assays. Vaccinia-specific CD8(+)/CFSE(lo)/IFN- gamma (+) and granzyme A(+) T cell responses were significantly correlated with the results of standard (51)Cr-release cytolytic assays (P.05). Furthermore, vaccinia induced antigen-specific memory gamma delta T cells. We demonstrate that vaccinia induces robust memory effector CD4(+), CD8(+), and gamma delta T cells, all of which are relevant for protection against smallpox. CFSE-based flow-cytometric assays will be useful in evaluating cell-mediated immune responses induced by new smallpox vaccines. |
| Databáze: | OpenAIRE |
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