Aiming allogeneic γδ T cells with tumor-specific antibodies via bioorthogonal chemistry for enhanced solid tumor immunotherapy

Background The development of "off-the-shelf" allogeneic γδ T-cell therapies is a major goal in cancer treatment. Current approaches, often reliant on genetic engineering, face challenges in manufacturing complexity, safety, and versatility for multi-targeting. Methods We developed a platform for constructing allogeneic antibody-γδ T cell conjugates using a bioorthogonal strategy based on metabolic glycan labeling and click chemistry. Conjugates were characterized for antibody attachment, specificity, and anti-tumor function. Results Conjugation was efficient and specific. Anti-Her2 antibody-conjugated γδ T cells (αHer2-γδ T) showed enhanced binding and selective clustering against Her2-positive tumor cells in vitro. αHer2-γδ T mediated significantly superior cytotoxicity compared to unconjugated controls, which was associated with increased activation marker (CD69/CD25) expression and IFN-γ release. In vivo, αHer2-γδ T exhibited improved tumor-homing and significant anti-tumor efficacy in xenograft models. The platform's broad applicability was demonstrated by generating functional conjugates targeting Trop-2 (αTrop-2-γδ T) and EGFR (αEGFR-γδ T), which also displayed potent, antigen-specific cytotoxicity against various solid tumors. Conclusion We have established a robust and versatile platform for creating "off-the-shelf" allogeneic antibody-γδ T cell conjugates. This strategy effectively equips γδ T cells with specific tumor-targeting capabilities, markedly enhancing their anti-tumor potency and presenting a promising, generalizable approach for next-generation allogeneic cell therapies.