Department of Microbiology, Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA 01843, USA.
J Immunol Methods. 2020 Sep-Oct;484-485:112830. doi: 10.1016/j.jim.2020.112830. Epub 2020 Aug 1.
Since the FDA approval of two Chimeric Antigen Receptor (CAR) T cell therapies against CD19 malignancies, there has been significant interest in adapting CAR technology to other diseases. As such, the ability to simultaneously monitor manufacturing criteria and functional characteristics of multiple CAR T cell products by a single instrument would likely accelerate the development of candidate therapies. Here, we demonstrate that image-based cytometry yields high-throughput measurements of CAR T cell proliferation and size, and captures the kinetics of in vitro antigen-specific CAR T cell-mediated killing. The data acquired and analyzed by the image cytometer are congruent with results derived from conventional technologies when tested contemporaneously. Moreover, the use of bright-field and fluorescence microscopy by the image cytometer provides kinetic measurements and rapid data acquisition, which are direct advantages over industry standard instruments. Together, image cytometry enables fast, reproducible measurements of CAR T cell manufacturing criteria and effector function, which can greatly facilitate the evaluation of novel CARs with therapeutic potential.
自 FDA 批准两种针对 CD19 恶性肿瘤的嵌合抗原受体 (CAR) T 细胞疗法以来，人们对将 CAR 技术应用于其他疾病产生了浓厚的兴趣。因此，能够通过单一仪器同时监测多种 CAR T 细胞产品的制造标准和功能特性，可能会加速候选疗法的开发。在这里，我们证明基于图像的细胞术可以高通量测量 CAR T 细胞的增殖和大小，并捕获体外抗原特异性 CAR T 细胞介导的杀伤动力学。通过图像细胞仪获取和分析的数据与同时进行的传统技术得出的结果一致。此外，图像细胞仪通过明场和荧光显微镜提供动力学测量和快速数据采集，这是相对于行业标准仪器的直接优势。总之，图像细胞术能够快速、可重复地测量 CAR T 细胞的制造标准和效应功能，这极大地促进了具有治疗潜力的新型 CAR 的评估。
