Adoptive transfer of gamma delta T cells is a novel immunotherapeutic approach to glioblastoma. Few recent studies have shown the efficacy of gamma delta T cells against glioblastoma, but no previous studies have identified the ligand-receptor interactions between gamma delta T cells and glioblastoma cells. Here, we identify those ligand-receptor interactions and provide a basis for using gamma delta T cells to treat glioblastoma. V gamma 9V delta 2 T cells were generated from peripheral blood mononuclear cells of healthy donors using artificial antigen presenting cells. MICA, ULBP, PVR and Nectin-2 expression in 10 patient-derived glioblastoma (PDG) cells were analyzed. The in vitro cytokine secretion from the gamma delta T cells and their cytotoxicity toward the PDG cells were also analyzed. The in vivo anti-tumor effects were evaluated using a U87 orthotopic xenograft glioblastoma model. Expression of ligands and cytotoxicity of the gamma delta T cells varied among the PDG cells. IFN-gamma and Granzyme B secretion levels were significantly higher when gamma delta Tcells were co-cultured with high-susceptible PDG cells than when they were co-cultured with low-susceptible PDG cells. Cytotoxicity correlated significantly with the expression levels of DNAM-1 ligands of the PDG cells. Blocking DNAM-1 resulted in a decrease in gamma delta T cell-mediated cytotoxicity and cytokine secretion. Intratumoral injection of gamma delta T cells showed anti-tumor effects in an orthotopic mouse model. Allogenic gamma delta T cells showed potent anti-tumor effects on glioblastoma in a DNAM-1 axis dependent manner. Our findings will facilitate the development of clinical strategies using gamma delta T cells for glioblastoma treatment.