T cells expressing chimeric antigen receptor promote immune tolerance
Antonio Pierini, Bettina P. Iliopoulou, Heshan Peiris, Magdiel Pérez-Cruz, Jeanette Baker, Katie Hsu, Xueying Gu, Ping-Ping Zheng, Tom Erkers, Sai-Wen Tang, William Strober, Maite Alvarez, Aaron Ring, Andrea Velardi, Robert S. Negrin, Seung K. Kim, Everett H. Meyer
Antonio Pierini, Bettina P. Iliopoulou, Heshan Peiris, Magdiel Pérez-Cruz, Jeanette Baker, Katie Hsu, Xueying Gu, Ping-Ping Zheng, Tom Erkers, Sai-Wen Tang, William Strober, Maite Alvarez, Aaron Ring, Andrea Velardi, Robert S. Negrin, Seung K. Kim, Everett H. Meyer
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Research Article Immunology Transplantation

T cells expressing chimeric antigen receptor promote immune tolerance

Abstract

Cellular therapies based on permanent genetic modification of conventional T cells have emerged as a promising strategy for cancer. However, it remains unknown if modification of T cell subsets, such as Tregs, could be useful in other settings, such as allograft transplantation. Here, we use a modular system based on a chimeric antigen receptor (CAR) that binds covalently modified mAbs to control Treg activation in vivo. Transient expression of this mAb-directed CAR (mAbCAR) in Tregs permitted Treg targeting to specific tissue sites and mitigated allograft responses, such as graft-versus-host disease. mAbCAR Tregs targeted to MHC class I proteins on allografts prolonged islet allograft survival and also prolonged the survival of secondary skin grafts specifically matched to the original islet allograft. Thus, transient genetic modification to produce mAbCAR T cells led to durable immune modulation, suggesting therapeutic targeting strategies for controlling alloreactivity in settings such as organ or tissue transplantation.

Authors

Antonio Pierini, Bettina P. Iliopoulou, Heshan Peiris, Magdiel Pérez-Cruz, Jeanette Baker, Katie Hsu, Xueying Gu, Ping-Ping Zheng, Tom Erkers, Sai-Wen Tang, William Strober, Maite Alvarez, Aaron Ring, Andrea Velardi, Robert S. Negrin, Seung K. Kim, Everett H. Meyer

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Figure 2

mAbCAR-expressing T cells are activated by FITC binding.

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mAbCAR-expressing T cells are activated by FITC binding. (A) Analysis of...
(A) Analysis of mAbCAR-expressing CD4+ T cells. Whiskers plots represent the percentage of surface expression of selected markers measured by flow cytometry (CD69, LAG3, PD1) before or after in vitro exposure to FITC (24 hours) in untransfected CD4+ T cells (red), CD4+ T cells that were mock transfected (black), and CD4+ T cells that were transfected with mAbCAR construct (gray). Reported results derive from 3 independent experiments. Two-tailed Student’s t test; mean ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001. (B) Activation of mAbCAR T cells in vitro is target specific. mAbCAR T cells loaded with FITC-isotype control mAb or FITC-anti-MAdCAM1 mAb and cultured for 1 day with irradiated cell suspension derived from syngeneic spleen where MAdCAM1 was also expressed. CD25 and CD69 surface expression was measured by flow cytometry. Reported results derive from 3 independent experiments. Two-tailed Student’s t test; mean ± SEM; *P < 0.05; **P < 0.01. (C) FITC mAbCAR does not modify T cell distribution. Percentage of naive (CD62L+CD44neg), central memory (CD62L+CD44+), and effector memory (CD62LnegCD44+) assessed by flow cytometry from mAbCAR T cells cultured as in B. Reported results derive from 3 independent experiments. Two-tailed Student’s t test; mean ± SEM. (D) Representative flow cytometric plot of naive (CD62L+CD44neg), central memory (CD62L+CD44+), and effector memory (CD62LnegCD44+) in vitro–cultured FITC-isotype or FITC-MAdCAM1-mAbCAR T cells.