Regulation of intestinal immunity and tissue repair by enteric glia (Single-cell expression analysis of tunica muscularis cells from naïve and H. poly-infected Ifngr2CTRL and Ifngr2ΔEGC mice.)

Tissue maintenance and repair depend on the integrated activity of multiple cell types. Whereas the contributions of epithelial, immune and stromal cells in intestinal tissue integrity are well understood, the role of intrinsic neuroglia networks remains largely unknown. Here, we uncover pivotal roles of enteric glial cells (EGCs) in intestinal homeostasis, immunity and tissue repair. We demonstrate that infection of mice with Heligmosomoides polygyrus (H. poly) leads to enteric gliosis and upregulation of the interferon gamma (IFN-γ) gene signature. Single-cell transcriptomics of tunica muscularis (TM) showed that glia-specific abrogation of IFN-γ signaling leads to tissue-wide activation of pro-inflammatory transcriptional programs. In addition, disruption of the IFN-γ-EGC signaling axis enhanced the inflammatory and granulomatous response of TM to helminths. Mechanistically, we show that upregulation of Cxcl10 is an early immediate response of EGCs to IFN-γ signaling and provide evidence that this chemokine and the downstream amplification of IFN-γ signaling in the TM are required for a measured inflammatory response to helminths and resolution of granulomatous pathology. Our study demonstrates that IFN-γ signaling in enteric glia is central to intestinal homeostasis and reveals critical roles of the IFN-γ-EGC-Cxcl10 axis in immune response and tissue repair following infectious challenge. To understand mechanistically how the IFN-γ-EGC axis regulates the intestinal response to helminth infection, we used scRNAseq analysis of the TM as an unbiased means to identify tissue-wide changes in cellular composition and gene expression associated with glia-specific abrogation of IFN-γ signalling. To understand the homeostatic roles of the IFN-γ-EGC signalling axis on the TM and to examine the role of the IFN-γ-EGC signalling axis in the response of host tissue to helminths, we compared the cellular and transcriptional landscape of the TM from naive and H. poly-infected Ifngr2CTRL (Sox10-CreERT2;Ifngr2+/fl, Ifngr2tm1.1Lflu, MGI: 6402706 15) and Ifngr2ΔEGC (Sox10-CreERT2;Ifngr2fl/fl) mice. We isolated cells from two experimental replicates (1: one mouse per sample, 2: a pool of three mice per sample) for each condition (naïve Ifngr2CTRL, naïve Ifngr2ΔEGC, 7 days of H. poly infection Ifngr2CTRL, 7 days of H. poly infection Ifngr2 ΔEGC) from both males and females at 10 days after tamoxifen induction.