IFN-γR/STAT1 signaling in recipient hematopoietic antigen-presenting cells suppresses graft-versus-host disease
Caisheng Lu, … , Suzanne Lentzsch, Markus Y. Mapara
Caisheng Lu, … , Suzanne Lentzsch, Markus Y. Mapara
Published November 29, 2022
Citation Information: J Clin Invest. 2023;133(3):e125986. https://doi.org/10.1172/JCI125986.
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Research Article Immunology

IFN-γR/STAT1 signaling in recipient hematopoietic antigen-presenting cells suppresses graft-versus-host disease

Abstract

The absence of IFN-γ receptor (IFN-γR) or STAT1 signaling in donor cells has been shown to result in reduced induction of acute graft-versus-host disease (GVHD). In this study, we unexpectedly observed increased activation and expansion of donor lymphocytes in both lymphohematopoietic organs and GVHD target tissues of IFN-γR/STAT1–deficient recipient mice, leading to rapid mortality following the induction of GVHD. LPS-matured, BM-derived Ifngr1–/– Stat1–/– DCs (BMDCs) were more potent allogeneic stimulators and expressed increased levels of MHC II and costimulatory molecules. Similar effects were observed in human antigen-presenting cells (APCs) with knockdown of Stat1 by CRISPR/Cas9 and treatment with a JAK1/2 inhibitor. Furthermore, we demonstrated that the absence of IFN-γR/STAT1 signaling in hematopoietic APCs impaired the presentation of exogenous antigens, while promoting the presentation of endogenous antigens. Thus, the indirect presentation of host antigens to donor lymphocytes was defective in IFN-γR/STAT1–deficient, donor-derived APCs in fully donor chimeric mice. The differential effects of IFN-γR/STAT1 signaling on endogenous and exogenous antigen presentation could provide further insight into the roles of the IFN-γ/STAT1 signaling pathway in the pathogenesis of GVHD, organ rejection, and autoimmune diseases.

Authors

Caisheng Lu, Huihui Ma, Liangsong Song, Hui Wang, Lily Wang, Shirong Li, Stephen M. Lagana, Antonia R. Sepulveda, Kasper Hoebe, Samuel S. Pan, Yong-Guang Yang, Suzanne Lentzsch, Markus Y. Mapara

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Figure 3

Absence of IFN-γR/STAT1 promotes an allostimulatory capacity in DCs.

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Absence of IFN-γR/STAT1 promotes an allostimulatory capacity in DCs. (A)...
(A) On day 1 after BMT, different APC populations from recipient animals were analyzed following induction of GVHD using a fully MHC-mismatched [BALB/c (H2d) to 129Sv (H2b)] strain combination. MHC II expression on recipient splenic H2b+ CD11c+, CD11b+, and B220+ cells from recipient 129.Stat1+/+ and 129. Stat1–/– animals. Results from 1 representative experiment of 3 are shown, with 3 animals/group. (B) GVHD was induced in the fully MHC-mismatched BALB/c (H2d) to B6 (H2b) strain combination in B6 WT or B6.Ifngr1–/– mice. I-Ab expression on recipient splenic CD11c+ cells was studied on day 1 after BMT. Results from 1 representative experiment of 2 are shown with 3–4 animals/group. (C) I-Ab and CD86 expression on CD11c+ cells from Ifngr1–/–→B6.SJL chimeric recipients were compared with expression on cells from the B6→B6.SJL counterparts 2 days after the second transplantation of BALB/c TCD BMCs and BALB/c-Luc splenocytes. Results from 1 representative experiment of 2 are shown, with 2–3 animals per group. (D) Expression of I-Ab, CD86, and PD-L1 in CD11c+ BMDCs from Stat1+/+ or Stat1–/– mice. Cells were cultured in RPMI 1640 medium containing 10% FCS and GM-CSF (20 ng/mL) for 6 days and matured in 100 ng/mL LPS for an additional 48 hours. Results from 1 representative experiment of more than 3 independent experiments are shown. (E–G) Proliferation and activation of CFSE-labeled alloreactive pan–T cells isolated from BALB/c splenocytes stimulated with LPS-matured Stat1+/+ or Stat1–/– BMDCs. Freshly isolated pan–T cells from BALB/c mice cells were stimulated for 5 days with LPS-matured Stat1+/+ or Stat1–/– BMDCs at a DC/responder ratio of 1:5. The proliferation of responder cells was assessed by 3H-incorporation, presented as the cpm ratio compared with unstimulated responder cells (E), or by CFSE dilution, presented as the percentage of CFSElo population (F). T cell activation was measured by CD44 and CD62L expression in responder CD4+ or CD8+ T cells (G). Results from 1 representative experiment of 3 independent experiments are shown. Bar graphs represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, by 2-way ANOVA with Šidák’s correction.