Human MHC class I chain-related A (MICA) is a tumor-associated antigen that can be recognized by Vdelta1 subset of tumor-infiltrating gammadelta T cells. We previously reported that immobilized recombinant MICA protein could induce the proliferation of tumor-infiltrating Vdelta1 gammadelta T cells in vitro. But there has been no direct evidence showing the engagement of gammadelta T cell receptors (TCR) of the induced cells with MICA. In the current investigation, we show that MICA induces specific cytolytic activity of the expanded gammadelta T cells. We expressed the coupled V domains from the MICA-induced T cells as a single polypeptide chain Vdelta Vgamma TCR (gammadelta scTCR). Such scTCR can specifically bind MICA of HeLa cells. Direct interaction of gammadelta scTCRs with in vitro expressed MICA was monitored using an IAsys biosensor. We found that the Vdelta1 scTCR can specifically bind to immobilized MICA molecule and MICA alpha1alpha2 domains are responsible for the binding reaction.