We examined whether γδ T and αβ T cells accumulating in early B16 melanoma lesions regulate NK and NK T cells that attack tumor cells. Freshly isolated and cultured tumor-infiltrating lymphocyte (TIL) populations of NK and NK T cells lysed B16 and produced IFN-γ, whereas γδ T and a large part of αβ T cell populations had no substantial cytotoxicity against B16 and secreted Th2 cytokines. Furthermore, the freshly isolated NK1. 1+ TIL population exhibited a higher anti-B16 effect than did splenocytes. γδ T and αβ T cell populations dramatically inhibited the cytotoxicity of NK and NK T cells in an MHC K b-dependent manner. Culture supernatant from γδ T and αβ T cell populations inhibited the proliferation of NK and NK T cell populations but did not affect their cytotoxicity, suggesting that the released Th2 cytokines are merely partly involved in the down-modulation of NK-lineage cells. NK1. 1+ cells obtained from TIL of γδ T cell-depleted mice significantly lysed B16 cells compared with those from control mice. Finally, anti-K b Fab mAb injected intralesionally at an early, but not at a late, stage of development of B16 melanoma inhibited tumor growth. These findings suggest that Th2-type γδ T and αβ T cells infiltrating in early B16 development inhibit the tumoricidal activity of NK-lineage cells using their class I molecules and partly their suppressive cytokines.