Interferon-gamma ELISPOT assay for the quantitative measurement of antigen-specific murine CD8+ T-cells.
Effective screening of new vaccines and immunotherapeutics requires assay methods that can provide quantitative measurement of cellular immune responses. The enzyme-linked immunospot (ELISPOT) is a sensitive technique for the detection of cytokine-producing cells at the single cell level. This assay is rapid and reproducible and permits the direct enumeration of low-frequency antigen-specific T-cells. This protocol describes in detail an interferon (IFN)-gamma ELISPOT method for measuring antigen-specific murine CD8+ T-cells. Spleen cells from specific cytotoxic T-lymphocyte (CTL) peptide-primed mice are used source of CD8+ T-cells to demonstrate the utility of this technique. The assay procedure is facilitated by the use of a ready-to-use IFN-gamma ELISPOT assay kit and it can be adapted to other model systems in which CD8+ T-cell responses are monitored.
Cole
Geoffrey A
GA
Point Therapeutics Inc., Boston, MA, USA.
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000008:administration & dosage / Q000737:chemistry; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002469:Cell Separation; D003627:Data Interpretation, Statistical; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005260:Female; D016056:Immunodominant Epitopes / Q000008:administration & dosage / Q000737:chemistry; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000032:analysis* / Q000096:biosynthesis*; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000941:Antigens; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D007371:Interferon-gamma