Lymphocytes infiltrating ovarian malignant ascites (TALs) were propagated in vitro in the presence of low (20-50 U/ml) concentrations of recombinant interleukin-2 (rIL-2) and moderate concentrations of interleukin-4 (IL-4) and tumor necrosis factor type alpha (TNF-alpha). Neither IL-4 nor TNF-alpha alone induced proliferation of ovarian TALs, but IL-4 and TNF-alpha synergized with IL-2 in enhancing the proliferation of T cells from cultures and inducing a more specific pattern of cytotoxic responses of ovarian TAL. IL-4 enhanced the proliferation of TAL cultures in the induction phase, but did not synergize with IL-2 in enhancing the proliferation and cytotoxicity of an established autologous tumor-specific CD8+/CD4- cytotoxic T lymphocyte (CTL) line. In contrast, TNF-alpha-induced preferential expansion of CD8+ CTL in TAL cultures where CD3+/CD4+ cells were initially in excess and of an established autologous tumor-specific CD8+ CTL line. This expansion resulted in increased levels of cytoxicity against autologous tumor cells. Amplification of IL-2 growth-promoting effects and selective enrichment in T cell subsets by IL-4 and TNF-alpha may therefore be useful in cellular adoptive immunotherapy using tumor-specific T cells.