One of the primary effector functions of immune cells is the killing of virus-infected or\nmalignant cells in the body. Natural killer (NK) and CD8 effector T cells are specialized for this\nfunction. The gold standard for measuring such cell-mediated cytolysis has been the chromium\nrelease assay, in which the leakage of the radioactive isotope from damaged target cells is being\ndetected. Flow cytometry-based single cell analysis of target cells has recently been established\nas a non-radioactive alternative. Here we introduce a target cell visualization assay (TVA) that\napplies similar target cell staining approaches as used in flow cytometry but based on single cell\ncomputer image analysis. Two versions of TVA are described here. In one, the decrease in numbers\nof calcein-stained, i.e., viable, target cells is assessed. In the other, the CFSE/PI TVA, the increase\nin numbers of dead target cells is established in addition. TVA assays are shown to operate with\nthe same sensitivity as standard chromium release assays, and, leaving data audit trails in form of\nscanned (raw), analyzed, and quality-controlled images, thus meeting requirements for measuring\ncell-mediated cytolysis in a regulated environment.
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