Cytotoxic Reactions of Murine Lymphoid Cells Studied With a Tritiated Proline Microcytotoxicity Test2

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Abstract
A lymphocyte cytotoxicity (CTX) test with 3H-proline-prelabeled target cells was used to detect the immune response of murine lymphoid cells to H-2 and tumor antigens. The specificity of the reaction was determined by simultaneous tests on unrelated target cells and, for reactions directed against H-2 antigens, by blocking experiments with alloantiserum directed against the H-2 antigens of the target cells. After a single intraperitoneal (ip) injection of allogeneic spleen cells, CTX of unfractionated peritoneal cells was strong, with a sharp peak on day 5. Repeated ip immunization markedly increased the CTX of unfractionated peritoneal cells. The reaction was strongest when the test was done at 37° C. Sometimes CTX could be detected after as little as 6 hours' incubation. CTX depended primarily on the absolute number of effector or target cells per area rather than on the ratio of effector to target cells. Both nonadherent and adherent peritoneal cells destroyed target cells specifically. The CTX of nonadherent peritoneal cells was increased by 2-mercaptoethanol. The CTX reaction depended on effector cells bearing Thy-1. Destruction of “innocent bystander” target cells was seen with one of four combinations of unfractionated and nonadherent peritoneal cells from hyperimmune animals.

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