γδ T细胞，以其既具有先天免疫系统，又具有后天免疫系统的特性，在癌症的细胞免疫治疗中是合适的候选者。由于它们的非主要组织相容性复合物（MHC）结合T细胞受体，可以进行异基因转移而没有相关的移植物抗宿主反应。近年来，通过使用双膦酸盐和白细胞介素2对γδ T细胞进行离体扩增和刺激已经积累了丰富经验。然而，许多当前的刺激方案都是基于使用异种材料和其他潜在危险的补充剂，与良好生产规范（GMP）的基本原则相矛盾。遵守GMP的概念和当前的指南是生产细胞治疗产品（ATMP）如细胞疗法的最新技术水平，并且对于在规章制度的视角下进行临床使用是必要的。在本研究中，我们开发了一种新的刺激方案，能够显著增加γδ T细胞数量，并使其更容易从研究过渡到临床应用，并减少规管工作量。它可可可可可可。我们的研究发现γδ T细胞纯度超过90％，与我们先前的标准流程相比，改善了体外抗肿瘤活性。此外，通过研究未刺激的γδ T细胞的特性与刺激的γδ T细胞的增殖率和脱颗粒能力之间的相关性，我们可以对合适的供体得出结论。最后，我们研究了使用唑来磷酸盐脉冲和/或使用白细胞介素15与或不使用白细胞介素2是否可以改善扩增。在内源性和抗体依赖的细胞介导细胞毒性方面取得了显著的改善。我们的结果表明，这里提出的刺激方案导致了改进的γδ T细胞产品，适用于未来的临床应用。©2023 Bold, Gross, Holzmann, Knop, Hoeres and Wilhelm 版权所有。

γδ T cells, with their properties of both the innate and acquired immune systems, are suitable candidates for cellular immunotherapy in cancer. Because of their non-major histocompatibility complex (MHC) binding T cell receptor, allogenic transfer is feasible without relevant graft versus host reactions. In recent years, much experience has been gained with ex vivo expansion and stimulation of γδ T cells using bisphosphonates and Interleukin 2. Unfortunately, many current stimulation protocols are based on the use of xenogenic materials and other potentially hazardous supplements, which conflicts with basic principles of Good Manufacturing Practice (GMP). Adherence to the concept and current guidelines of GMP is state of the art for production of Advanced Therapy Medicinal Products (ATMP) like cell therapeutics and a necessity for clinical use under a regulatory perspective. In this study, we developed a new stimulation protocol that induces a marked increase of γδ T cell counts and allows for an easier transition from research to clinical applications with minimized regulatory workload. It reliably leads to a cell product with a purity of more than 90% γδ T cells and improved in vitro anti-tumor activity compared to our previous standard procedure. Furthermore, by investigating correlations between properties of unstimulated γδ T cells and proliferation rate as well as degranulation ability of stimulated γδ T cells, we can draw conclusions about suitable donors. Finally, we examined if expansion can be improved by pulsing zoledronate and/or using Interleukin 15 with or without Interleukin 2. Significant improvements can be achieved with respect to intrinsic and antibody-dependent cell-mediated cytotoxicity. Our results demonstrate that the stimulation protocol presented here leads to an improved γδ T cell product for future clinical applications.Copyright © 2023 Bold, Gross, Holzmann, Knop, Hoeres and Wilhelm.