Characterization of CAR T cell expansion and cytotoxic potential during Ex Vivo manufacturing using image-based cytometry.

Autor: Maldini CR; Department of Microbiology, Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA., Love AC; Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA 01843, USA., Tosh KW; Department of Microbiology, Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA., Chan LL; Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA 01843, USA., Gayout K; Department of Microbiology, Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA., Smith T; Department of Technology R&D, Nexcelom Bioscience LLC, Lawrence, MA 01843, USA., Riley JL; Department of Microbiology, Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: rileyj@upenn.edu.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2020 Sep - Oct; Vol. 484-485, pp. 112830. Date of Electronic Publication: 2020 Aug 01.
DOI: 10.1016/j.jim.2020.112830
Abstrakt: 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.
Competing Interests: Declaration of Competing Interest JLR has filed a patent describing the construction of these HIV-specific CARs. JLR co-founded Tmunity Therapeutics that has the rights to license the CAR technology described herein. JLR holds an equity interest in Tmunity. AL, LLC, and TS are employees of Nexcelom Bioscience LLC. and declare competing financial interests. The research in this manuscript is for reporting on a biological detection method using the Celigo Image Cytometer, a product of Nexcelom Bioscience LLC. Authors declare no other conflicts of interest.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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