Wang Dongrui, Yang Xin, Xella Agata, Stern Lawrence A, Brown Christine E
T Cell Therapeutics Research Laboratories, Cellular Immunotherapy Center, Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, CA, United States.
Department of Chemical, Biological and Materials Engineering, University of South Florida, Tampa, FL, United States.
Methods Cell Biol. 2023;173:173-189. doi: 10.1016/bs.mcb.2022.07.010. Epub 2022 Sep 6.
The effector potency of chimeric antigen receptor (CAR) T cell therapeutic products is essential to their clinical antitumor responses, and potency monitoring is a critical quality control method for CAR T cell therapy platforms. While many in vitro assays enable high-throughput assessment of CAR T cell cytotoxicity, it has been challenging for these assays to reflect the in vivo therapeutic effect due to their nature as short-term methods that fail to recapitulate the high tumor burden environment. Here, we describe two in vitro co-culture methods to evaluate CAR T cell recursive killing potential at high tumor cell loads. In these assays, long-term cytotoxic function and proliferative capacity of CAR T cells are examined in vitro over 7days. Further, these assays are coupled with profiling CAR T cell expansion, cytokine production and phenotypes. These methods provide a facile approach to assess CAR T cell potency and to elucidate the functional variations across different CAR T cell products.
嵌合抗原受体（CAR）T细胞治疗产品的效应效力对其临床抗肿瘤反应至关重要，效力监测是CAR T细胞治疗平台的关键质量控制方法。虽然许多体外试验能够高通量评估CAR T细胞的细胞毒性，但由于这些试验本质上是短期方法，无法重现高肿瘤负荷环境，因此难以反映体内治疗效果。在此，我们描述了两种体外共培养方法，用于评估高肿瘤细胞负荷下CAR T细胞的递归杀伤潜力。在这些试验中，在体外7天内检测CAR T细胞的长期细胞毒性功能和增殖能力。此外，这些试验还结合了对CAR T细胞扩增、细胞因子产生和表型的分析。这些方法为评估CAR T细胞效力和阐明不同CAR T细胞产品之间的功能差异提供了一种简便的方法。
