Human gammadelta T cells expressing a V gamma 9V delta 2 T-cell receptor (TCR) kill various tumour cells including autologous tumours. In addition to TCR-dependent recognition, activation of NKG2D-positive gammadelta T cells by tumour cell-expressed NKG2D ligands can also trigger cytotoxic effector function. In this study, we investigated the involvement of TCR versus NKG2D in tumour cell recognition as a prerequisite to identify tumour types suitable for gammadelta T-cell-based immunotherapy. We have characterized epithelial tumour cells of different origin with respect to cell surface expression of the known NKG2D ligands MHC class I-chain-related antigens (MIC) A/B and UL16-binding proteins (ULBP), and susceptibility to gammadelta T-cell killing. Most tumour cells expressed comparable levels of MICA and MICB as well as ULBP with the exception of ULBP-1 which was absent or only weakly expressed. Most epithelial tumours were susceptible to allogeneic gammadelta T-cell lysis and in the case of an established ovarian carcinoma to autologous gammadelta T-cell killing. Lysis of resistant cells was enhanced by pre-treatment of tumour cells with aminobisphosphonates or pre-activation of gammadelta T cells with phosphoantigens. A potential involvement of TCR and/or NKG2D was investigated by antibody blockade. These experiments revealed three patterns of inhibition, i.e. preferential inhibition by anti-TCR antibody, preferential inhibition by anti-NKG2D antibody, or additive blockade by anti-TCR plus anti-NKG2D antibodies. Our results indicate for the first time that the NKG2D pathway is involved in the lysis of different melanomas, pancreatic adenocarcinomas, squamous cell carcinomas of the head and neck, and lung carcinoma.