Rorc restrains the potency of ST2+ regulatory T cells in ameliorating intestinal graft-versus-host disease
Jinfeng Yang, … , Bruce R Blazar, Sophie Paczesny
Jinfeng Yang, … , Bruce R Blazar, Sophie Paczesny
Published January 29, 2019
Citation Information: JCI Insight. 2019;4(5):e122014. https://doi.org/10.1172/jci.insight.122014.
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Research Article Immunology Transplantation

Rorc restrains the potency of ST2+ regulatory T cells in ameliorating intestinal graft-versus-host disease

Abstract

Soluble stimulation–2 (ST2) is increased during graft-versus-host disease (GVHD), while Tregs that express ST2 prevent GVHD through unknown mechanisms. Transplantation of Foxp3– T cells and Tregs that were collected and sorted from different Foxp3 reporter mice indicated that in mice that developed GVHD, ST2+ Tregs were thymus derived and predominantly localized to the intestine. ST2–/– Treg transplantation was associated with reduced total intestinal Treg frequency and activation. ST2–/– versus WT intestinal Treg transcriptomes showed decreased Treg functional markers and, reciprocally, increased Rorc expression. Rorc–/– T cells transplantation enhanced the frequency and function of intestinal ST2+ Tregs and reduced GVHD through decreased gut-infiltrating soluble ST2–producing type 1 and increased IL-4/IL-10–producing type 2 T cells. Cotransfer of ST2+ Tregs sorted from Rorc–/– mice with WT CD25-depleted T cells decreased GVHD severity and mortality, increased intestinal ST2+KLRG1+ Tregs, and decreased type 1 T cells after transplantation, indicating an intrinsic mechanism. Ex vivo IL-33–stimulated Tregs (TregIL-33) expressed higher amphiregulin and displayed better immunosuppression, and adoptive transfer prevented GVHD better than control Tregs or TregIL-33 cultured with IL-23/IL-17. Amphiregulin blockade by neutralizing antibody in vivo abolished the protective effect of TregIL-33. Our data show that inverse expression of ST2 and RORγt in intestinal Tregs determines GVHD and that TregIL-33 has potential as a cellular therapy avenue for preventing GVHD.

Authors

Jinfeng Yang, Abdulraouf Ramadan, Dawn K. Reichenbach, Michael Loschi, Jilu Zhang, Brad Griesenauer, Hong Liu, Keli L. Hippen, Bruce R Blazar, Sophie Paczesny

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Figure 3

Rorc–/– ST2+ Tregs decrease GVHD severity and mortality, increase intestinal ST2+ KLRG1+ Tregs after HCT, and decrease intestinal type 1 T cells.

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Rorc–/– ST2+ Tregs decrease GVHD severity and mortality, increase intest...
(A) Clinical scores and the survival curve of lethally irradiated C3H.SW mice that received B6 WT TCD BM and WT CD25-depleted total T cells with 10% ST2+ Tregs isolated from WT or Rorc–/– mice. n = 7, from 2 independent experiments, data are shown as mean ± SEM; unpaired t test, *P < 0.05 for clinical scores; a log-rank test was used for survival analysis, **P < 0.01. (B) Representative plots of ST2 expression on Tregs isolated from the gut of C3H.SW mice in A receiving WT or Rorc–/– ST2+ Tregs analyzed on day 14 after allo-HCT. n = 4, data are shown as mean ± SEM; unpaired t test, *P < 0.05. (C) Representative plots of ST2, KLRG1, and Helios expression on Foxp3+ Tregs from the gut of C3H.SW mice in A receiving WT or Rorc–/– ST2+ Tregs on day 14 after HCT. n = 4, data are shown as mean ± SEM; unpaired t test, *P < 0.05. (D) Total infiltrated cells in the gastrointestinal (GI) tract of mice in A receiving WT or Rorc–/– ST2+ Tregs analyzed on day 14 after HCT. n = 4, data are shown as mean ± SEM; unpaired t test, *P < 0.05. (E) Representative plots of IFN-γ production by CD4+ and CD8+ Teffs from the gut of C3H.SW mice in A receiving WT or Rorc–/– ST2+ Tregs analyzed on day 14 after allo-HCT. n = 4, data are shown as mean ± SEM; unpaired t test, **P < 0.01, ***P < 0.001.