Abstract 6288: A high-throughput in vitro characterization method of CAR T cells using image cytometry

Abstract

Abstract Since the FDA approval of two CAR T cell therapies, Kymriah (Tisagenlecleucel, Novartis) and Yescarta (Axicabtagene ciloleucel, Kite) for CD19+ malignancies, there has been a significant interest for developing new CARs targeting other disease indications. As such, the ability to simultaneously monitor manufacturing criteria and functional characteristics of multiple CAR T cell products would likely be critical to accelerate candidate therapies into clinical trials. Here, we demonstrate that image-based cytometry using the Celigo Image Cytometer (Nexcelom Bioscience LLC) yields high-throughput measurements of CAR T cell proliferation and size throughout the cell-manufacturing process, and captures the kinetics of antigen-specific CAR T cell-mediated killing in vitro. The data acquired and analyzed by the Celigo Image Cytometer were congruent with results from conventional instruments when tested contemporaneously, such as the Multisizer 3 (Beckman Coulter) which monitored the properties of CAR T cell expansion, and a luciferase-based assay which measured the cytotoxic potential of CAR T cells. More importantly, the use of bright field and fluorescent imaging by the Celigo Image Cytometer provided kinetic measurements and rapid data acquisition and analysis which were direct advantages over the conventional technologies described herein. Together, the Celigo Image Cytometer enabled fast, reproducible, and high-throughput assessment of CAR T cell manufacturing criteria and effector function, which could greatly facilitate the identification and evaluation of novel CARs with therapeutic potential. Citation Format: Leo Li-Ying Chan, Colby Maldini, Andrea Love, Kevin Tosh, Kevin Gayout, Tim Smith, James Riley. A high-throughput in vitro characterization method of CAR T cells using image cytometry [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6288.