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Generation of Cytotoxic Lymphocytes in Vitro: Response of Immune Rat Spleen Cells to a Syngeneic Gross Virus-Induced Lymphoma in Mixed Lymphocyte-Tumor Culture¹

From the Division of Tumor Immunology, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104; and the Departments of Pediatrics, Microbiology, and Medicine, University of Washington School of Medicine, Seattle, Washington 98195

Abstract

Spleen cells from W/Fu rats 4 to 6 weeks after immunization with syngeneic Gross virus-induced lymphoma (C58NT)D cells usually lack detectable activity in a short-term ⁵¹Cr release assay. The results presented here demonstrate that these spleen cells retain the capacity to generate significant proliferative and cytotoxic activity upon re-exposure to mitomycin C-treated (C58NT)D cells in vitro. Optimal conditions were defined in W/Fu rats for this secondary immune response in vitro to the (C58NT)D cells. The cytotoxic response was observed to be quantitative, reproducible, and specific. Optimal generation occurred 5 days after initiation of cultures with a 30:1 responding cell:stimulating cell ratio. In vitro generated cytotoxic cells inhibit tumor growth in vivo when administered as a mixture with tumor cells.

Footnotes

¹ This investigation was supported by Grants CA-16712 and CA-17481 awarded by the National Cancer Institute.