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Reduced intestinal epithelial mitochondrial function enhances in vitro interleukin-8 production in response to commensal Escherichia coli

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Abstract

Uncoupling of oxidative phosphorylation in epithelial mitochondria results in decreased epithelial barrier function as characterized by increased internalization of non-invasive Escherichia coli and their translocation across the epithelium. We hypothesized that the increased burden of intracellular commensal bacteria would activate the enterocyte, with the potential to promote inflammation. Treatment of human colon-derived epithelial cell lines in vitro with dinitrophenol (DNP) and commensal E. coli (strains F18, HB101) provoked increased production of interleukin (IL-8), which was not observed with conditioned medium from the bacteria, lipopolysaccharide or inert beads. The IL-8 response was inhibited by co-treatment with cytochalasin-D (blocks F-actin rearrangement), chloroquine (blocks phagosome acidification) and a MyD88 inhibitor (blocks TLR signaling), consistent with TLR-signaling mediating IL-8 synthesis subsequent to bacterial internalization. Use of the mitochondria-targeted antioxidant, mitoTEMPO, or U0126 to block ERK1/2 MAPK signalling inhibited DNP+E. coli-evoked IL-8 production. Mutations in the NOD2 (the intracellular sensor of bacteria) or ATG16L1 (autophagy protein) genes are susceptibility traits for Crohn’s, and epithelia lacking either protein displayed enhanced IL-8 production in comparison to wild-type cells when exposed to DNP + E coli. Thus, metabolic stress perturbs the normal epithelial–bacterial interaction resulting in increased IL-8 production due to uptake of bacteria into the enterocyte: this potentially pro-inflammatory event is enhanced in cells lacking NOD2 or ATG16L1 that favor increased survival of bacteria within the enterocyte. We speculate that by increasing epithelial permeability and IL-8 production, reduced mitochondria function in the enteric epithelium would contribute to the initiation, pathophysiology, and reactivation of inflammatory disease in the gut.

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Acknowledgements

A. Saxena was supported by a Queen Elizabeth II Graduate Studentship Award. F. Lopes was supported by CIHR/Canadian Association of Gastroenterology (CAG)/Allergan Inc. and Alberta Innovates-Health solutions (AI-HS) Post-Doctoral Fellowships. D.M. McKay holds a Canada Research Chair (CRC: Tier 1) in Intestinal Immunophysiology in Health and Disease.

Funding

Funding provided by a Canadian Institutes for Health Research (CIHR) Operating Grant (MPO-126005) to D.M. McKay.

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  1. Gastrointestinal Research Group and Inflammation Research Network, Department of Physiology and Pharmacology, Calvin, Joan and Phoebe Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, 1877 HSC, 3330 Hospital Drive NW, Calgary, AB, T2N 4N1, Canada

    Alpana Saxena, Fernando Lopes & Derek M. McKay

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  1. Alpana Saxena
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Correspondence to Derek M. McKay.

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Saxena, A., Lopes, F. & McKay, D.M. Reduced intestinal epithelial mitochondrial function enhances in vitro interleukin-8 production in response to commensal Escherichia coli. Inflamm. Res. 67, 829–837 (2018). https://doi.org/10.1007/s00011-018-1172-5

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