In vivo and in vitro induction of cytotoxic lymphocytes by alloantigen-reactive lymphocytes fractionated on spleen cell monolayers

The generation of cytotoxic T cells by normal and primed lymphocytes fractionated on spleen cell monolayers was examined by two methods. In the first method, sublethally irradiated recipients were inoculated with the fractionated cells, and 5 to 6 days later the spleens were tested for cytotoxicity against ⁵¹Cr tumor target cells syngeneic to the host. Significant reduction of cytotoxic activity was observed in one-half of the experiments. In the second method, the fractionated cells were cultured with mitomycin C-treated stimulated cells and tested 6 days later for cytotoxicity. A slight reduction of cytotoxic activity was observed. In both methods, the cytotoxic response generated by the fractionated alloantigen-primed lymphocytes was similar to the response obtained with unprimed cells.