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| Article Authors: | X S He; B Rehermann; J Boisvert; J Mumm; H T Maecker; M Roederer; T L Wright; V C Maino; M M Davis; H B Greenberg |
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| Article Title: | Direct functional analysis of epitope-specific CD8+ T cells in peripheral blood. |
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| Reference ID: | 1001709 |
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| Abstract: | The functional status of virus-specific CD8+ T cells is important for the outcome and the immunopathogenesis of viral infections. We have developed an assay for the direct functional analysis of antigen-specific CD8+ T cells, which does not require prolonged in vitro cultivation and amplification of T cells. Whole blood samples were incubated with peptide antigens for <5 h, followed by staining with peptide-MHC tetramers to identify epitope-specific T cells. The cells were also stained for the activation marker CD69 or for the production of cytokines such as interferon-gamma (IFNgamma) or tumor necrosis factor-alpha (TNFalpha). With the combined staining with tetramer and antibodies to CD69 or cytokines the number of antigen-specific CD8+ T cells as well as the functional response of each individual cell to the cognate antigen can be determined in a single experiment. Virus-specific CD8+ T cells that are nonfunctional, as well as those that are functional under the same stimulating conditions can be simultaneously detected with this assay, which is not possible by using other T-cell functional assays including cytotoxicity assay, intracellular cytokine staining, and enzyme-linked immunospot (ELISPOT) assay. |
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| Affiliations: | Department of Medicine, Stanford University School of Medicine, California 94305-5187, USA. xiaosong@stanford.edu |
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| Date: | 2001 |
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| Reference Type: | Literature |
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| PubMed ID: | 11270597 |
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| Journal: | Viral Immunol |
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| Journal Volume: | 14 |
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| Article Pages: | 59-69 |
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| Journal ISSN: | 0882-8245 |
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| Article Chemical List: | Antigens, Viral;Epitopes, T-Lymphocyte;Peptides;Receptors, Antigen, T-Cell;Phycoerythrin |
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| Article MeSH List: | Antigens, Viral(immunology); CD8-Positive T-Lymphocytes(immunology); Cytomegalovirus(immunology); Cytomegalovirus Infections(immunology); Epitopes, T-Lymphocyte(immunology); Hepacivirus(immunology); Hepatitis C(immunology); Humans; Lymphocyte Activation(immunology); Major Histocompatibility Complex(physiology); Peptides(metabolism); Phycoerythrin(metabolism); Receptors, Antigen, T-Cell(metabolism); Staining and Labeling |
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| Curation Last Updated: | 2013-05-28 20:21:43 |
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