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Enriching leukapheresis improves T cell activation and transduction efficiency during CAR T processing

Noaks, E; Peticone, C; Kotsopoulou, E; Bracewell, DG; (2021) Enriching leukapheresis improves T cell activation and transduction efficiency during CAR T processing. Molecular Therapy - Methods and Clinical Development , 20 pp. 675-687. 10.1016/j.omtm.2021.02.002. (In press). Green open access

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Abstract

The majority of CD19-directed CAR T cell products are manufactured using an autologous process. Although using a patient's leukapheresis reduces the risks of rejection, it introduces variability in starting material composition and the presence of cell populations that might negatively affect production of chimeric antigen receptor (CAR) T cells, such as myeloid cells. In this work, the effect of monocytes (CD14) on the level of activation, growth, and transduction efficiency was monitored across well plate and culture bag platforms using healthy donor leukapheresis. Removal of monocytes from leukapheresis improved the level of activation 2-fold, achieving the same level of activation as when initiating the process with a purified T cell starting material. Two activation reagents were tested in well plate cultures, revealing differing sensitivities to starting material composition. Monocyte depletion in culture bag systems had a significant effect on transduction efficiency, improving consistency and increasing the level of CAR expression by up to 64% compared to unsorted leukapheresis. Cytotoxicity assays revealed that CAR T cell products produced from donor material depleted of monocytes and isolated T cells consistently outperformed those made from unsorted leukapheresis. Analysis of memory phenotypes and gene expression indicated that CAR T cells produced using depleted starting material displayed a more rested and naive state. The success of CAR T cell manufacturing and final product function is influenced by the composition of the donor starting material. In this work, we show that upstream depletion of specific cell populations can enhance processing outcomes such as activation, transduction, and phenotype of the therapeutic product.

Type: Article
Title: Enriching leukapheresis improves T cell activation and transduction efficiency during CAR T processing
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.omtm.2021.02.002
Publisher version: http://dx.doi.org/10.1016/j.omtm.2021.02.002
Language: English
Additional information: © 2021 The Author(s). 675This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: CAR T-cell, cell therapy, manufacture, enrichment, monocytes, activation, transduction, leukapheresis
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10123750
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