Document Type : Original Article
Department of Zoology, Faculty of Science, Tanta University, Tanta, Egypt
Clinical Pathology, Faculty of Medicine, Tanta University, Tanta, Egypt.
Department of Zoology, Faculty of Science, Damanhour University, Elbehira, Egypt
Background: In a series of preclinical studies, we have reported that IL-12 can enhance the phenotype and functionality of CD8+ T cells during their in vitro expansion for anti-cancer adoptive T cell therapy (ACT). Vγ9 +Vδ2+ T cells can be used in ACT since they are expanded and activated upon their stimulation with aminobisphosphonates such as zoledronate (ZOL). Aim: We aimed in this study to utilize IL-12 to enhance the expansion and functions of ZOL/ IL-2-expanded Vγ9 +Vδ2+ T cells as well as CD8+ T cells from breast cancer patients in comparison to healthy control subjects upon the condition in vitro. Materials and Methods: Peripheral blood mononuclear cells (PBMCS) were separated from healthy donors (HD) and stage II breast cancer patients (BCII). PBMCs were cultured (1x106 cells/mL) in a complete RPMI medium, and treated with combinations of ZOL+IL-2, ZOL+IL-2+IL-12 or IL-2+IL-12. Cultured cells were harvested on days 7 and 14 of culture and their phenotypic analysis and cytolytic function were performed by flow cytometry. Results: The addition of IL-12 during expansion by ZOL+IL-2 promoted the responsiveness of expanded Vγ9+Vδ2+ T cells from breast cancer patients significantly as ZOL+IL-2 effect. Interestingly, ZOL+IL-12+IL-2 induced a robust granzyme B (GZB) and perforin (PER) production from Vγ9+Vδ2+ at D7 of the stimulation. Conclusion: ZOL+IL-2+IL-12 effect on cytolytic activity from Vγ9+Vδ2+ was instantly at D7 and gradual increase of CD8+ cytolytic activity at D14. Which supported a balanced activation level during the expansion process.