Automated colorimetric assay for T cell cytotoxicity

Abstract

A colorimetric method has been developed for detecting the lysis of target cells by cytotoxic T lymphocytes (Tc). The method entails incubating Tc cells with thioglycollate-induced macrophage targets and estimating macrophage survival at the end of the assay by staining viable macrophages with the dye neutral red. The method is substantially more sensitive than the 51Cr release assay and can be used to detect alloreactive Tc cells and H-2-restricted Tc cells against viruses, haptens and minor-H antigens. Furthermore, the assay is applicable to limit dilution analysis of Tc cell precursors. The method is cheap, avoids radioactive materials and by measuring optical densities with automated spectrophotometers developed for microELISA systems, results can be obtained 50-100 times faster than with the radioactive procedure.