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Study of the Cellular Immune Response to Gross Virus-induced Lymphoma by the Mixed Lymphocyte-Tumor Interaction

Cellular and Tumor Immunology Section, Laboratory of Cell Biology, National Cancer Institute, Bethesda, Maryland 20014

A one-way mixed lymphocyte-tumor cell interaction assay was applied to the study of cell-mediated immunity to a syngeneic Gross virus-induced lymphoma, (C58NT)D, in W/Fu rats. Lymphoid cells from the spleens of rats inoculated by the tumor cells were stimulated in vitro by incubation with intact, inactivated tumor cells. Optimal conditions for stimulation were incubation of 6 x 10⁵ spleen cells and 2 x 10⁴ mitomycin C-treated tumor cells for 4 days. Significant stimulation was first detected at 14 days after tumor cell inoculation, reached a peak at Days 20 to 40, and gradually declined to low levels by Day 90. Immunization with from 1 x 10⁵ to 1 x 10⁸ tumor cells induced similar reactivity; 1 x 10⁹ cells produced progressive tumor growth and no reactivity was seen. Treatment of immune spleen cells with specific anti-rat thymocyte serum abolished their response while removal of complement receptor-bearing lymphocytes from the spleens did not reduce the reactivity. Specificity was shown by inability of three other W/Fu rat tumor cells to stimulate (C58NT)D-immune W/Fu spleen cells and by inability of (C58NT)D cells to stimulate spleen cells obtained either from normal W/Fu rats or from W/Fu rats inoculated with another W/Fu tumor.

These results indicate that the one-way mixed lymphocyte tumor cell interaction may measure cellular immunity and not primary recognition of antigens of Gross virus-induced lymphoma and that thymus-processed lymphoid cells are needed for the reactivity in the one-way mixed lymphocyte-tumor cell interaction.

Received 2/ 1/74. Accepted 5/ 7/74.