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Generation of non-human primate CAR Tregs using artificial antigen-presenting cells, simian tropic lentiviral vectors, and antigen-specific restimulation

Gavin I. Ellis, Mosha Z. Deng, Delaine W. Winn and 6 more

STAR Protocols · 2022

doi:10.1016/j.xpro.2022.101784 (opens in a new tab)

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Abstract

Summary: It is technically challenging to generate large doses of regulatory T cells (Tregs) engineered to express a chimeric antigen receptor (CAR) in non-human primates (NHP). Here, we have optimized the manufacturing of CAR Tregs by stringent sorting of Tregs, stimulation by artificial antigen-presenting cells, transduction by simian tropic lentiviral vectors, and antigen-specific expansion. The result of this method is highly suppressive CAR Tregs for use in a pre-clinical, large animal model of transplant tolerance.For complete details on the use and execution of this protocol, please refer to Ellis et al. (2022).: Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract licence: CC BY-NC

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