Human allogenic γδ T cells kill patient-derived glioblastoma cells expressing high levels of DNAM-1 ligands

Abstract

Adoptive transfer of γδ T cells is a novel immunotherapeutic approach to glioblastoma. Few recent studies have shown the efficacy of γδ T cells against glioblastoma, but no previous studies have identified the ligand–receptor interactions between γδ T cells and glioblastoma cells. Here, we identify those ligand–receptor interactions and provide a basis for using γδ T cells to treat glioblastoma. Vγ9Vδ2 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 γδ 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 γδ T cells varied among the PDG cells. IFN-γ and Granzyme B secretion levels were significantly higher when γδ 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 γδ T cell–mediated cytotoxicity and cytokine secretion. Intratumoral injection of γδ T cells showed anti-tumor effects in an orthotopic mouse model. Allogenic γδ 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 γδ T cells for glioblastoma treatment.

B7H3-targeting chimeric antigen receptor modification enhances antitumor effect of Vγ9Vδ2 T cells in glioblastoma

“…Vγ9Vδ2 T cell immunotherapy offers a promising approach. In this study, we demonstrated that Vγ9Vδ2 T cells derived from healthy human PBMCs inhibited GBM cell proliferation in vitro and suppressed GBM tumor growth in vivo without apparent toxicity to mice, consistent with recent reports [ 35 – 37 ]. However, the response to Vγ9Vδ2 T cell therapy varied greatly among PTCs, leading us to classify them into SAT and WAT groups.…”

CD155/PVR determines acute myeloid leukemia targeting by Delta One T cells

“…We first focused on DNAM-1 and its ligands, PVR and nectin-2, emerging players in tumor immunology, 11 previously shown to be important for Vδ2 + γδ T-cell responses to liver or brain cancers. 12 , 13 , 14 We used blocking antibodies in 3-hour killing assays against HEL or MOLM13 AML cells, and found a 20% to 30% decrease in DOT-cell cytotoxicity upon DNAM-1 neutralization ( Figure 1 D-E). However, because ligand blockade failed to reproduce this effect, we used a more stringent methodology, CRISPR–CRISPR-associated protein 9 gene editing, to knock out each or both ligands ( Figure 1 F).…”

Isolation and expansion of pure and functional γδ T cells

“…The mode of action of gd T cells in an anti-tumor response comprises direct cytolytic activity via NKG2D, FcyRIII, Fas/Fasligand, DNAM-1 receptor-ligand interaction and the TNF related apoptosis inducing ligand (TRAIL) pathway resulting in production of perforins, granzymes, IFN-g and TNF-a (4,5,(18)(19)(20)(21)(22)(23). In concurrence, gd T cells can also activate other immune cells (24,25), for instance activate NK cells via co-stimulation, promote dendritic cell (DC) maturation and support initiation of a humoral immune response (26)(27)(28)(29)(30)(31).…”