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.