Analyzing CD8 T cells in mouse models of poxvirus infection.
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.
Research School of Biology, The Australian National University, Canberra, ACT, Australia.
Inge E A
IE
Flesch
Yik Chun
YC
Wong
David C
DC
Tscharke
D000595:Amino Acid Sequence; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism*; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic; D004195:Disease Models, Animal; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D053804:Granzymes / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism; D008198:Lymph Nodes / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000473:pathology; D013194:Staining and Labeling; D014615:Vaccinia / Q000276:immunology* / Q000473:pathology; D014616:Vaccinia virus
D018984:Epitopes, T-Lymphocyte; D019041:L-Selectin; D007371:Interferon-gamma; D053804:Granzymes; C509214:Gzmb protein, mouse