Analysis of frequency and phenotype of antigen-specific T cells.
Over the last decade, our understanding of the cellular immune system has been greatly advanced through the development of methods to identify antigen-specific T cells directly ex vivo. The major reagents and techniques used for this purpose are (i) tetramerised MHC:peptide complexes (tetramers) which bind to specific T-cell receptors (TCR) and (ii) assays that detect T cells which synthesise cytokines in response to cognate stimulation (intracellular cytokine staining (ICS)). Here, we provide a detailed description of the procedure for generating and using class I MHC:peptide tetramers to label peptide-specific T cells and for carrying out ICS to measure antigen-specific T lymphocytes.
Stock
Angus
A
Nuffield Department of Clinical Medicine, Weatherall Institute of Molecular Medicine, Oxford, UK.
Cerundolo
Vincenzo
V
D000818:Animals; D000941:Antigens / Q000276:immunology*; D016207:Cytokines / Q000378:metabolism; D000939:Epitopes / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000302:isolation & purification; D007158:Immunologic Techniques; D042541:Intracellular Space / Q000378:metabolism; D018655:Lymphocyte Count / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D020836:Protein Structure, Quaternary; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology*
D000941:Antigens; D016207:Cytokines; D000939:Epitopes; D015395:Histocompatibility Antigens Class I; D010455:Peptides