Loss of IL-10 signaling in macrophages limits bacterial killing driven by prostaglandin E2.

Subhankar Mukhopadhyay,Tobias Schwerd,Immacolata Porreca,Christine Hale,Gordon Dougan,Simon Travis,William C Skarnes,Luke Jostins-Dean,Jessica L Forbester,Yoon Ha Choi,Kaur Alasoo,Chukwuma A Agu,Fiona Powrie,Melania Capitani,Holm H Uhlig,Amy Yeung,Eva Heinz,Julia Rodrigues,David Thomas ,Daniel J Gaffney ,Haiying Yang ,Nicholas R Thomson

doi:10.1084/jem.20180649

Abstract

Loss of IL-10 signaling in macrophages (Mφs) leads to inflammatory bowel disease (IBD). Induced pluripotent stem cells (iPSCs) were generated from an infantile-onset IBD patient lacking a functional IL10RB gene. Mφs differentiated from IL-10RB-/- iPSCs lacked IL-10RB mRNA expression, were unable to phosphorylate STAT3, and failed to reduce LPS induced inflammatory cytokines in the presence of exogenous IL-10. IL-10RB-/- Mφs exhibited a striking defect in their ability to kill Salmonella enterica serovar Typhimurium, which was rescuable after experimentally introducing functional copies of the IL10RB gene. Genes involved in synthesis and receptor pathways for eicosanoid prostaglandin E2 (PGE2) were more highly induced in IL-10RB-/- Mφs, and these Mφs produced higher amounts of PGE2 after LPS stimulation compared with controls. Furthermore, pharmacological inhibition of PGE2 synthesis and PGE2 receptor blockade enhanced bacterial killing in Mφs. These results identify a regulatory interaction between IL-10 and PGE2, dysregulation of which may drive aberrant Mφ activation and impaired host defense contributing to IBD pathogenesis.

Highlights

The inflammatory bowel diseases (IBDs) encompassing Crohn’s disease and ulcerative colitis are complex chronic inflammatory conditions of the gastrointestinal tract
We used four independent human Induced pluripotent stem cells (iPSCs) lines (HIPSI0114i-kolf_2, HPSI0813fpdj_3, HPSI0314i-bubh_1, and HPSI0713i-uimo_1) from unrelated healthy individuals obtained from the Human Induced Pluripotent Stem Cell Initiative
The IL-10RB−/− and the four control iPSC lines were differentiated into Mφs (Alasoo et al, 2015; Hale et al, 2015) with very similar phenotypes in terms of morphology and expression of markers

Summary

Introduction

The inflammatory bowel diseases (IBDs) encompassing Crohn’s disease and ulcerative colitis are complex chronic inflammatory conditions of the gastrointestinal tract. Mouse models identified IL-10 as a critical cytokine in the maintenance of intestinal homeostasis (Kole and Maloy, 2014; Kühn et al, 1993; Moore et al, 2001) through limiting the activation state of macrophages (Mφs; Bogdan et al, 1991; Smythies et al, 2005). Myeloid cell–specific loss of IL-10 receptor α (Shouval et al, 2014a; Zigmond et al, 2014), IL-10 receptor β (Donnelly et al, 2004; Kotenko et al, 1997; Spencer et al, 1998), or STAT3 (Kobayashi et al, 2003) all lead to spontaneous colitis in mice, suggesting that IL-10 signaling in myeloid cells is critical for the maintenance of intestinal homeostasis. In the absence of IL-10 signaling, the resident microbiota drives excessive MyD88-dependent TLR signaling and uncontrolled Mφ activation, contributing to intestinal pathology (Hoshi et al, 2012; Rakoff-Nahoum et al, 2006; Redhu et al, 2017)

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Results

Conclusion