Activation of autoreactive lymphocytes in the lung by radioresistant cells expressing a STING gain-of-function mutation
Kevin MingJie Gao, Kristy Chiang, Sharon Subramanian, Xihui Yin, Paul J. Utz, Kerstin Nündel, Kate A. Fitzgerald, Ann Marshak-Rothstein
Kevin MingJie Gao, Kristy Chiang, Sharon Subramanian, Xihui Yin, Paul J. Utz, Kerstin Nündel, Kate A. Fitzgerald, Ann Marshak-Rothstein
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Research Article Inflammation

Activation of autoreactive lymphocytes in the lung by radioresistant cells expressing a STING gain-of-function mutation

Abstract

Gain-of-function mutations in the dsDNA sensing adaptor STING lead to a severe autoinflammatory syndrome known as STING-associated vasculopathy with onset in infancy (SAVI). Patients with SAVI develop interstitial lung disease (ILD) and produce autoantibodies that are commonly associated with systemic autoimmune diseases. Mice expressing the most common SAVI mutation, STING V154M (VM), similarly develop ILD but exhibit severe T and B cell lymphopenia and low serum Ig titers, and they lack autoantibodies. Importantly, lethally irradiated VM hosts reconstituted with WT stem cells (WT→VM) still develop ILD. In this study, we find that WT→VM chimeras had restored B cell function, produced autoantibodies, and thereby recapitulated the loss of tolerance seen in patients with SAVI. Lymphocytes derived from both WT and BCR or TCR transgenic (Tg) donors accumulated in the extravascular lung tissue of WT+Tg→VM mixed chimeras, but lymphocyte activation and germinal center formation required WT cells with a diverse repertoire. Furthermore, when T cells isolated from the WT→VM chimeras were adoptively transferred to naive Rag1-deficient secondary hosts, they trafficked to the lung and recruited neutrophils. Overall, these findings indicated that VM expression by radioresistant cells promoted the activation of autoreactive B cells and T cells that then differentiated into potentially pathogenic effector subsets.

Authors

Kevin MingJie Gao, Kristy Chiang, Sharon Subramanian, Xihui Yin, Paul J. Utz, Kerstin Nündel, Kate A. Fitzgerald, Ann Marshak-Rothstein

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Figure 7

WT lymphocytes from WT→VM chimeras induce lung inflammation in Rag1-KO mice.

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WT lymphocytes from WT→VM chimeras induce lung inflammation in Rag1-KO m...
(A) Six-week-old CD45.2/.2 WT and VM mice were lethally irradiated and reconstituted with CD45.1/1 WT donor BM to generate WT→WT and WT→VM chimeric mice. Splenocytes from these chimeric mice were then harvested 8 weeks after BM engraftment and adoptively transferred into secondary recipient 8-week-old Rag1–/– recipients to generate (WT→WT)→Rag1–/– (n = 9) and (WT→VM)→Rag1–/– (n = 11) mice and then evaluated 8–9 weeks later. (B) Representative H&E lung histology from (WT→WT)→Rag1–/– and (WT→VM)→Rag1–/– mice captured with a 4× objective. A region of interest, as indicated by dotted red box, was further imaged with a 10× objective and shown 5× magnified over the original image. Data in B are representative of n = 9 (WT→VM)→Rag1–/– mice and n = 11 (WT→WT)→Rag1–/– mice. (C) Lungs from (WT→WT)→Rag1–/– and (WT→VM)→Rag1–/– mice, stained for DAPI (gray), CD3 (yellow), B220 (cyan), and LYVE-1 (magenta). Data in C are representative of n = 3 (WT→VM)→Rag1–/– mice and n = 2 (WT→WT)→Rag1–/– mice. (D) Total number of EV T and B cells in the lungs of (WT→WT)→Rag1–/– and (WT→VM)→Rag1–/– mice. (E) Percentage of donor-derived cells in the CD11b+ or CD11c+ lung EV myeloid compartment. (F) Percentage of CD11b+Ly6G+ neutrophils from the host and donor lung myeloid cell compartment. (G) Number of neutrophils from host lung EV myeloid compartment. Nonparametric Mann-Whitney U tests were used for pair-wise comparisons to determine statistical significance (*P < 0.05, **P < 0.01). Scale bar: 200 μm.