摘　要：γδT细胞是一类表达γδT细胞受体的T淋巴细胞亚群，具有先天免疫和适应性免疫功能，能够独立于MHC限制直接识别并杀伤肿瘤细胞。然而，其在体内数量较少，且体外扩增效率低，限制了其临床应用。本研究使用细胞因子白介素-2（IL-2）、白介素-15（IL-15）和双膦酸类药物唑来膦酸（Zol）联合诱导刺激γδT细胞，以优化其体外扩增策略。结果表明，Zol（5 μmol/L）、IL-2（100 IU/mL）和IL-15（10 ng/mL）联合使用可使γδT细胞扩增10 000倍，并且γδT细胞在CD3阳性细胞中的纯度达到95.74%。此外，二甲双胍处理显著提高了γδT细胞的中央记忆型细胞亚群的比例（10.75%提高至15.85%）和效应记忆型细胞亚群的比例（5.98%提高至19.3%）（P<0.01）。二甲双胍显著上调γδT细胞抗肿瘤因子干扰素-γ（IFN-γ）、颗粒酶B（GZMB）和穿孔素（perforin）的表达，促进了γδT细胞分泌外泌体，增强了其细胞毒性作用。本研究为γδT细胞的体外扩增及功能优化提供了新策略，并为其在肿瘤过继性免疫治疗中的应用奠定了基础。
关键词：γδT细胞；抗肿瘤作用；二甲双胍；抗肿瘤因子；过继性免疫治疗
中图分类号：Q254                           文献标志码：A                  DOI：10.3969/j.issn.1007-7146.2025.03.005

Abstract: Gamma delta (γδ) T cells are a subset of T lymphocytes expressing γδT cell receptors, which possess both innate and adaptive immune functions. They can recognize and kill tumor cells in a manner independently of MHC restriction. However, their limited quantity in vivo and low efficiency in vitro expansion restrict their clinical application. This study utilized a combination of cytokines interleukin-2 (IL-2), interleukin-15 (IL-15), and the bisphosphonate drug zoledronic acid (Zol) to stimulate γδT cells and optimize their in vitro expansion strategy. Results demonstrated that the combination of Zol (5 μmol/L), IL-2 (100 IU/mL), and IL-15 (10 ng/mL) achieved a 10 000-fold expansion of γδT cells, with a purity of 95.74% among CD3-positive cells. Additionally, metformin treatment significantly increased the proportions of central memory cell subsets from 10.75% to 15.85% and effector memory cell subsets from 5.98% to 19.3% in γδT cells (P<0.01). Metformin also markedly upregulated the expression of anti-tumor factors interferon-γ (IFN-γ), granzyme B (GZMB), and perforin in γδT cells, promoted exosome secretion, and enhanced cytotoxic activity. This study provides a novel strategy for in vitro expansion and functional optimization of γδT cells, laying the foundation for their application in adoptive immunotherapy for tumors.
Key words: γδT cells; antitumor activity; metformin; anti-tumor factor; adoptive immunotherapy
（Acta Laser Biology Sinica, 2025, 34(3): 229-238）