Analyzing CD8 T cells in mouse models of poxvirus infection

Methods Mol Biol. 2012:890:199-218. doi: 10.1007/978-1-61779-876-4_12.

Abstract

Mouse models of immunology are frequently used to study host responses to poxviruses or poxvirus-based recombinant vaccines. In this context, the magnitude of CD8(+) T cell responses is often of interest. Methods to evaluate CD8(+) T cell responses extend from those that rely on indirect measurement of effector function only, such as cytotoxicity assays, to those that only measure antiviral CD8(+) T cell numbers and not function, like peptide MHC tetramers. In this chapter, five methods are provided that cover this range: DimerX staining (a variant of peptide-MHC tetramers), intracellular cytokine staining for interferon-γ, CD62L/Granzyme B staining, and in vitro and ex vivo cytotoxicity assays. We also include tables of vaccinia virus peptide epitopes for use in most of these assays.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CD8-Positive T-Lymphocytes / metabolism*
  • Cell Line
  • Cytotoxicity Tests, Immunologic
  • Disease Models, Animal
  • Epitopes, T-Lymphocyte / metabolism
  • Female
  • Flow Cytometry
  • Granzymes / metabolism
  • Humans
  • Interferon-gamma / metabolism
  • L-Selectin / metabolism
  • Lymph Nodes / virology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Spleen / pathology
  • Staining and Labeling
  • Vaccinia / immunology*
  • Vaccinia / pathology
  • Vaccinia virus*

Substances

  • Epitopes, T-Lymphocyte
  • L-Selectin
  • Interferon-gamma
  • Granzymes
  • Gzmb protein, mouse