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The effect of Adriamycin on the generation of cell-mediated cytotoxicity in the mixed cell culture was studied in patients with various carcinomas. Peripheral blood mononuclear cells (PBM) from the patients were cultured with the B-lymphoblastoid cell line Raji in mixed culture, and the induced cytotoxicity was measured by 51Cr release assay. In patients with various carcinomas, the capacity of PBM to be converted to cytotoxic cells was significantly augmented 5, 7, and 10 days after a single dose of 25 mg/sq m i.v., when compared to that of PBM obtained before Adriamycin injection. The peak level of the cytotoxicity observed 7 days after injection was more than 2-fold higher than that before treatment. Although the depletion of adherent cells from PBM either before or after treatment resulted in a decreased cytotoxic response, nonadherent cell fractions as well as unfractionated cells from PBM after drug treatment equally showed an augmented response when compared to that before injection. The distribution of T-cell subsets exhibited a significant increase in the percentage of OKT8 positive cells after administration. Furthermore, PBM obtained after treatment produced significantly higher levels of interleukin 2. The results appear to indicate that the imbalance of T-cell subsets and the increase of interleukin 2 production may be related to the augmenting effect of Adriamycin administration on cytotoxic response in cancer patients.