In vitro detection of cell‐mediated immunity to individual tumor‐specific antigens of chemically induced BALB/c fibrosarcomas

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Abstract
In this study we have analyzed the in vitro cell‐mediated cytotoxicity of immune peritoneal exudate cells (PEC), elicited in syngeneic mice against the MCA‐induced, TSTA‐bearing BALB/c fibrosarcomas CA‐2, GI‐17 and C‐3. The 4 h 51Cr‐release assay showed the immune PEC effectors to be specifically cytotoxic to fibrosarcoma used for the immunization, but not to other syngeneic MCA‐induced tumors or normal fibroblasts. Cold target inhibition experiments on CA‐2 cells confirmed the specificity of the reaction. When PEC, lymph‐node and spleen cells from BALB/c anti‐CA‐2 mice were compared for anti‐tumor activity, only PEC were found to kill tumor cells significantly. PEC effectors did not have a significant level of NK or NC activities since they were unable to destroy YAC‐1 target and the PEC‐mediated anti‐tumor activity was not inhibited by unlabelted YAC‐1 or WEHI‐164 tumor cells. PEC anti‐CA‐2 were analyzed for the expression of T‐cell markers by anti‐Thy 1.2, anti‐Ly 1.2 and Ly 2.2 monoclonal antibodies. Anti‐tumor specific effector cells were identified as mature T cells since they were not adherent to plastic and showed Thy l.2+, Ly 1.2− and Ly 2.2+ phenotypes. In addition, anti‐H‐2Kd but not anti‐H‐2Dd alloantiserum added to target cells, blocked CA‐2 tumor lysis, thus supporting the conclusion that the T‐cell response against TSTA is H‐2 restricted.

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