Background: In a series of our preclinical studies, we have reported that conditioning of α/β CD8+ T cells in vitro with interleukin-12 (IL-12) during their expansion improves their homing phenotype and anti-tumor cytolytic function upon their adoptive transfer in vivo. Vγ9+Vδ2+ T cells can also be expanded in vitro with amino bisphosphonates such as zoledronate (ZOL) for the purpose of adoptive therapy. Aim: We aimed in this study to use IL-12 to enhance the expansion and cytotoxic functions of ZOL-expanded Vγ9+Vδ2+T cells. Materials and methods: Peripheral blood mononuclear cells (PBMCs) were separated from healthy donors and stage II breast cancer patients. PBMCs (1 × 106 cells/mL) were cultured and treated with ZOL/IL2, ZOL/IL2/IL12, or IL2/IL12. Cultured cells were harvested on days 7 and 14 of culture and their numbers, phenotype, and cytolytic activity were assessed. The levels of pro- and inflammatory cytokines/chemokines in the plasma and supernatants of the cultured cells were analyzed by Luminex. Results: In healthy subjects, the addition of IL-12 to ZOL/IL2-stimulated PBMCs increased the expansion and the cytotoxic activity of Vγ9+Vδ2+ T cells on days 7 and 14 of culture. The latter was measured by the expression level of the cytolytic molecules granzyme B (GZB) and perforin (PER). Of note, αβ CD8 + T cells were also activated under the same condition but with a lesser extent addition of IL-12 to ZOL/IL2-stimulated PBMCs from cancer patients also induced similar effects but were lower than in control subjects. Interestingly, ZOL/IL2/IL12-treated PBMCs showed higher levels of cytokines/chemokines, in particular, CCL, CCL4, GM-CSF, IL-1rα; IL-12, IL-13, TNF, and IFNγ measured on days 7 and 14. Conclusion: The addition of IL12 at the start of the expansion protocol can enhance the activity of γδ T cells which might be mediated in part by the activation of αβ T cells.Keywords: Chemokines; Cytokines; Granzyme B; IL-12; IL-2; Perforin; Zoledronate; αβ T cells; γδ T cells.

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乐备实（上海优宁维生物科技股份有限公司旗下全资子公司），是国内专注于提供高质量蛋白检测以及组学分析服务的实验服务专家，自2018年成立以来，乐备实不断寻求突破，公司的服务技术平台已扩展到单细胞测序、空间多组学、流式检测、超敏电化学发光、Luminex多因子检测、抗体芯片、PCR Array、ELISA、Elispot、PLA蛋白互作、多色免疫组化、DSP空间多组学等30多个，建立起了一套涵盖基因、蛋白、细胞以及组织水平实验的完整检测体系。

 
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