Phenotypic and Functional Analysis of Antigen-Specific T Cell Exhaustion

Methods Mol Biol. 2017:1514:83-92. doi: 10.1007/978-1-4939-6548-9_6.

Abstract

In some cancers and chronic infections exhausted T cells increase their expression of inhibitory receptors and demonstrate an impaired ability to produce cytokines and to proliferate. Immunological techniques such as MHC class I tetramer staining, intracellular cytokine staining, and CFSE dilution can be used to determine the memory status, inhibitory receptor expression, cytokine production, and proliferative capacity of antigen-specific CD8+ T cells. Here, we describe approaches to define the inhibitory receptor expression, cytokine production, and proliferative capacity of antigen-specific CD8+ T cells from HIV-infected and CMV-infected donors by using polychromatic flow cytometry.

Keywords: CMV; Carboxyfluorescein succinimidyl ester; Exhaustion; Flow cytometry; HIV; Inhibitory receptors; Intracellular cytokine staining; MHC class I tetramer; T cells.

MeSH terms

  • CD8-Positive T-Lymphocytes / immunology*
  • Cytokines / immunology
  • Flow Cytometry / methods*
  • Genes, MHC Class I / immunology
  • HIV Infections / immunology*
  • HIV Infections / pathology
  • Humans
  • Immunologic Techniques / methods*
  • Interferon-gamma / immunology

Substances

  • Cytokines
  • Interferon-gamma