Allograft rejection is restrained by short-lived TIM-3+PD-1+Foxp3+ Tregs
Shipra Gupta, … , Vijay K. Kuchroo, Terry B. Strom
Shipra Gupta, … , Vijay K. Kuchroo, Terry B. Strom
Published June 11, 2012
Citation Information: J Clin Invest. 2012;122(7):2395-2404. https://doi.org/10.1172/JCI45138.
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Research Article Immunology

Allograft rejection is restrained by short-lived TIM-3+PD-1+Foxp3+ Tregs

Abstract

Tregs play a pivotal role in inducing and maintaining donor-specific transplant tolerance. The T cell immunoglobulin and mucin domain-3 protein (TIM-3) is expressed on many fully activated effector T cells. Along with program death 1 (PD-1), TIM-3 is used as a marker for exhausted effector T cells, and interaction with its ligand, galectin-9, leads to selective death of TIM-3+ cells. We report herein the presence of a galectin-9–sensitive CD4+FoxP3+TIM-3+ population of T cells, which arose from CD4+FoxP3+TIM-3– proliferating T cells in vitro and in vivo and were often PD-1+. These cells became very prominent among graft-infiltrating Tregs during allograft response. The frequency and number of TIM-3+ Tregs peaked at the time of graft rejection and declined thereafter. Moreover, these cells also arise in a tolerance-promoting donor-specific transfusion model, representing a pool of proliferating, donor-specific Tregs. Compared with TIM-3– Tregs, TIM-3+ Tregs, which are often PD-1+ as well, exhibited higher in vitro effector function and more robust expression of CD25, CD39, CD73, CTLA-4, IL-10, and TGF-β but not galectin-9. However, these TIM-3+ Tregs did not flourish when passively transferred to newly transplanted hosts. These data suggest that a heretofore unrecognized graft-infiltrating, short-lived subset of Tregs can restrain rejection.

Authors

Shipra Gupta, Thomas B. Thornley, Wenda Gao, Rafael Larocca, Laurence A. Turka, Vijay K. Kuchroo, Terry B. Strom

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Figure 1

The number of CD4+FoxP3+TIM-3+ T cells within dLNs and spleens arising during the allograft response peak at graft rejection.

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The number of CD4+FoxP3+TIM-3+ T cells within dLNs and spleens arising d...
Fully allogeneic BALB/c (H-2d) or syngeneic C57BL/6 (H-2b) full-thickness body skin was grafted on the lateral thorax of FoxP3 GFP-KI reporter C57BL/6 (H-2b) mice. C57BL/6-KI mice reject the BALB/c skin graft on day 7. The number of FoxP3-GFP+TIM-3+ cells in (A) dLNs and (B) spleens from skin transplant recipients was determined by flow cytometry (n ≥ 8 mice). (A and B) The TIM-3+FoxP3+ T cells within dLNs and spleens peaked at the time of rejection and were more numerous in allogeneic as compared with syngeneic grafted hosts. Data are presented as mean ± SEM; *P < 0.05; **P < 0.01. (C) Gated CD4+GFP+FoxP3+ cells harvested from collagenase digested allogeneic skin transplants were analyzed for TIM-3 expression by flow cytometry. CD4+GFP+FoxP3+TIM-3+ Tregs comprised 40% of the graft-infiltrating Tregs on day 5 after transplantation. (D) The magnitude of TIM-3 expression on the TIM-3+ Tregs is depicted in comparison to the staining control. Gray shading represents fluorescence minus 1 for TIM-3 staining, and the black line represents TIM-3 expression on CD4+GFP+FoxP3+ cells in the graft (n = 4).