Interferon- 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)-γ 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-γ ELISPOT assay kit and it can be adapted to other model systems in which CD8⁺ T-cell responses are monitored.