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

STAR Protoc. 2022 Nov 5;3(4):101784. doi: 10.1016/j.xpro.2022.101784. eCollection 2022 Dec 16.

Abstract

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).

Keywords: Biotechnology and bioengineering; Cell culture; Cell isolation; Flow cytometry/Mass cytometry; Immunology; Model organisms; Tissue engineering.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen-Presenting Cells
  • Primates
  • Receptors, Chimeric Antigen* / genetics
  • T-Lymphocytes, Regulatory
  • Transplantation Tolerance

Substances

  • Receptors, Chimeric Antigen