Direct functional analysis of epitope-specific CD8+ T cells in peripheral blood

Xiaosong He, H. B. Greenberg, B. Rehermann, J. Boisvert, J. Mumm, H. T. Maecker, M. Roederer, T. L. Wright, V. C. Maino, M. M. Davis

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

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 (IFNγ) or tumor necrosis factor-alpha (TNFα). 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.

Original languageEnglish (US)
Pages (from-to)59-69
Number of pages11
JournalViral Immunology
Volume14
Issue number1
StatePublished - 2001
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Virology

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