Dendritic epidermal T cells (DETC), which are skin-specific members of the tissue-resident γδ T-cell family, are characterized by their potential to kill selected tumor targets by a non-major histocompatibility complex (MHC)-restricted mechanism. We have recently identified a new receptor molecule, 2B4, that appears to be associated with non-MHC-restricted recognition of tumor targets by natural killer cells. The purpose of this study was to determine whether DETC express 2B4 molecules, and, if so, to assess their functional roles in DETC-mediated killing of tumor targets. Short-term DETC lines as well as DETC freshly procured from skin expressed surface 2B4, as detected with a specific monoclonal antibody. Removal of interleukin (IL)-2 from DETC cultures caused substantial reduction in 2B4 expression levels as well as a reduction in cytotoxic capacity against YAC-1 targets in a standard 51Cr-release assay. Conversely, exposure to IL-2, but not to IL-7, elevated both 2B4 expression and cytotoxicity. To assess the functional roles played by surface 2B4, we pretreated DETC lines with anti-2B4 antibody and then tested for their killing potential. Anti-2B4, but not the control antibody, augmented their capacity to lyse YAC-1 targets (51Cr-release assays) and to disrupt the monolayers of Pam-212-transformed keratinocytes (visual assessment). Thus, we conclude that DETC express, in an IL-2-dependent manner, 2B4 molecules, which may play a unique role in the killing of skin-derived tumors.