The IFN-γ receptor promotes immune dysregulation and disease in STING gain-of-function mice
W. Alexander Stinson, Cathrine A. Miner, Fang R. Zhao, Annena Jane Lundgren, Subhajit Poddar, Jonathan J. Miner
W. Alexander Stinson, Cathrine A. Miner, Fang R. Zhao, Annena Jane Lundgren, Subhajit Poddar, Jonathan J. Miner
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Research Article Inflammation

The IFN-γ receptor promotes immune dysregulation and disease in STING gain-of-function mice

Abstract

STING gain-of-function mutations cause STING-associated vasculopathy with onset in infancy (SAVI) in humans, a disease characterized by spontaneous lung inflammation and fibrosis. Mice with STING gain-of-function mutations (SAVI mice) develop αβ T cell–dependent lung disease and also lack lymph nodes. Although SAVI has been regarded as a type I interferonopathy, the relative contributions of the three interferon receptors are incompletely understood. Here, we show that STING gain of function led to upregulation of IFN-γ–induced chemokines in the lungs of SAVI mice and that deletion of the type II IFN receptor (IFNGR1), but not the type I IFN receptor (IFNAR1) or type III IFN receptor (IFNλR1), ameliorated lung disease and restored lymph node development in SAVI mice. Furthermore, deletion of IFNGR1, but not IFNAR1 or IFNλR1, corrected the ratio of effector to Tregs in SAVI mice and in mixed bone marrow chimeric mice. Finally, cultured SAVI mouse macrophages were hyperresponsive to IFN-γ, but not IFN-β, in terms of Cxcl9 upregulation and cell activation. These results demonstrate that IFNGR1 plays a major role in autoinflammation and immune dysregulation mediated by STING gain of function.

Authors

W. Alexander Stinson, Cathrine A. Miner, Fang R. Zhao, Annena Jane Lundgren, Subhajit Poddar, Jonathan J. Miner

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Figure 6

IFNGR1 partially mediates STING-associated T cell proliferation and survival defects in cell culture.

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IFNGR1 partially mediates STING-associated T cell proliferation and surv...
(A) Representative FACS plots showing eYFP expression in splenic CD4+ and CD8α+ T cells from WT and STING N153S (SAVI) IFN-γ reporter (IfngeYFP) and nonreporter (Ifng+/+) mice. (B) CD4+ and CD8α+ T cell eYFP mean fluorescence intensity (MFI) in animals from A. Data represent the mean of n = 7–9 mice pooled from 2 independent experiments. (C) Percentage of T cells after 24-hour coculture that stained positively for the viability dye LIVE/DEAD Aqua. (D) Representative FACS plots depicting frequency of T cells isolated from mice of the indicated genotype following 24 hours in coculture. (E) Percentage distribution of isolated WT or SAVI T cells following 24-hour coculture. T cells were isolated from CD45.1 WT, CD45.2 SAVI, or CD45.2 WT animals, and data represent the mean cellular frequency of n = 5 samples pooled from 2 independent experiments. (F and G) Representative FACS plots depicting dilution of CFSE in and the resulting division indices ± SEM of CD4+ T cells (F) or CD8α+ T cells (G) following 3-day mock or α-CD3ε/α-CD28 stimulation of bulk splenocytes isolated from n = 5–6 mice of the indicated genotypes. (H and I) Percentage cell death of CD4+ (H) and CD8α+ (I) T cells following 3-day mock or α-CD3ε/α-CD28 stimulation of bulk splenocytes isolated from n = 5–6 mice of the indicated genotypes. (F–I) Data represent the mean pooled 3 independent experiments. (J) WT STING-expressing OTI CD8α+ T cell proliferation in response to WT and SAVI bone marrow–derived macrophages (BMDMs) loaded with OVA peptide. BMDMs were harvested loaded with OVA peptide under conditions of LPS stimulation, and then cultured with WT STING-expressing OTI T cells for 3 days. Data represent the mean of n = 6 samples pooled from 2 independent experiments. Data were analyzed by Student’s t test (B); 1-way ANOVA (C, E, and J); 2-way ANOVA (F–I). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.