CD19-targeted CAR regulatory T cells suppress B cell pathology without GvHD
Yuki Imura, … , Minako Ito, Akihiko Yoshimura
Yuki Imura, … , Minako Ito, Akihiko Yoshimura
Published June 11, 2020
Citation Information: JCI Insight. 2020;5(14):e136185. https://doi.org/10.1172/jci.insight.136185.
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Research Article Therapeutics

CD19-targeted CAR regulatory T cells suppress B cell pathology without GvHD

Abstract

Regulatory T cells (Tregs) play essential roles in maintaining immunological self-tolerance and preventing autoimmunity. The adoptive transfer of antigen-specific Tregs has been expected to be a potent therapeutic method for autoimmune diseases, severe allergy, and rejection in organ transplantation. However, effective Treg therapy has not yet been established because of the difficulty in preparing a limited number of antigen-specific Tregs. Chimeric antigen receptor (CAR) T cells have been shown to be a powerful therapeutic method for treating B cell lymphomas, but application of CAR to Treg-mediated therapy has not yet been established. Here, we generated CD19-targeted CAR (CD19-CAR) Tregs from human PBMCs (hPBMCs) and optimized the fraction of the Treg source as CD4+CD25+CD127loCD45RA+CD45RO–. CD19-CAR Tregs could be expanded in vitro while maintaining Treg properties, including high expression of the latent form of TGF-β. CD19-CAR Tregs suppressed IgG antibody production and differentiation of B cells via a TGF-β–dependent mechanism. Unlike conventional CD19-CAR CD8+ T cells, CD19-CAR Tregs suppressed antibody production in immunodeficient mice that were reconstituted with hPBMCs, reducing the risk of graft-versus-host disease. Therefore, the adoptive transfer of CD19-CAR Tregs may provide a novel therapeutic method for treating autoantibody-mediated autoimmune diseases.

Authors

Yuki Imura, Makoto Ando, Taisuke Kondo, Minako Ito, Akihiko Yoshimura

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Figure 1

Generation of CD19-targeted CAR Tregs.

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Generation of CD19-targeted CAR Tregs. (A) Schematic representation of c...
(A) Schematic representation of constructs carrying CD19-targeted CAR and empty vector. EC, extracellular domain; TM, transmembrane domain; CP, cytoplasmic domain. (B) Sorting strategy to isolate naive/resting CD45RA+ regulatory T cells (CD45RA+ Tregs), CD45RO+ Tregs, and conventional CD4 T cells (Tconvs). (C) Scheme for the generation and expansion of CD19-targeted CAR (CD19-CAR) Tregs and CD19-CAR Tconvs. (D) Flow cytometric analysis of CAR (Venus) when transduced cells were sorted on days 4–6. The left panels show representative FACS profiles, and the right panel shows quantification of the transduction rate (n = 23 or 26). Data are representative of 26 independent experiments using human samples that were provided by 5 healthy donors. (E) Fold expansion on days 12–14, 8 days after CAR T cells were sorted (n = 17, 14, or 12). Data are representative of 17 independent experiments using human samples provided by 5 healthy donors. P values were determined using (D) 2-tailed Student’s t test or (E) 1-way ANOVA (*P < 0.05, **P < 0.01). Data are presented as mean ± SEM.