Abstract
Clostridium difficile infection (CDI) is mediated by two major exotoxins, toxin A (TcdA) and toxin B (TcdB), that damage the colonic epithelial barrier and induce inflammatory responses. The function of the colonic vascular barrier during CDI has been relatively understudied. Here we report increased colonic vascular permeability in CDI mice and elevated vascular endothelial growth factor A (VEGF-A), which was induced in vivo by infection with TcdA- and/or TcdB-producing C. difficile strains but not with a TcdA−TcdB− isogenic mutant. TcdA or TcdB also induced the expression of VEGF-A in human colonic mucosal biopsies. Hypoxia-inducible factor signalling appeared to mediate toxin-induced VEGF production in colonocytes, which can further stimulate human intestinal microvascular endothelial cells. Both neutralization of VEGF-A and inhibition of its signalling pathway attenuated CDI in vivo. Compared to healthy controls, CDI patients had significantly higher serum VEGF-A that subsequently decreased after treatment. Our findings indicate critical roles for toxin-induced VEGF-A and colonic vascular permeability in CDI pathogenesis and may also point to the pathophysiological significance of the gut vascular barrier in response to virulence factors of enteric pathogens. As an alternative to pathogen-targeted therapy, this study may enable new host-directed therapeutic approaches for severe, refractory CDI.
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All data reported in this study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank W.-R. Lie (EMD Millipore) for help with the cytokine assays and T. Fei (Dana Farber Cancer Institute) for technical help. This work was supported by an Irving W. and Charlotte F. Rabb Award (to X.C.), Crohn’s and Colitis Foundation of America Career Development Award (to X.C.), Young Investigator Award for Probiotic Research (to X.C.), NIH/NIDDL grant no. P30 DK 41301 (to C.P.), NIH/NIDDK grant no. U01 DK110003 (to C.P.), NIH/NIAID R01 grant no. DK60729 (to C.P.), fellowship from the Crohn’s and Colitis Foundation of America (to K.B.), Foundation of China National Key Clinical Discipline (to J.W.), Foundation of China Scholarship Council (to J.Huang), National Health and Medical Research Council of Australia grant no. 1107812 (to D.L.), Australian Research Council Future Fellowship grant no. FT120100779 (to D.L.) and NIH/NIAID R01 grant nos AI116596 (to C.P.K.) and U19 AI 109776 (to C.P.K.).
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X.C., C.P.K., J.Huang, D.L., C.P. and J.W. were involved in study design, analysis and interpretation of data, drafted the manuscript and obtained funding. J.Huang, J.A.V.G., Y.M.S., K.B., H.X., X.Y., W.Z., Y.Z., J.D.G., S.J.M., S.L., D.P.-S., K.S.S., I.J.P. and J.Hansen contributed towards the acquisition of data. C.P., Y.M.S., A.C.M., M.H. and S.Y.-H. provided material support.
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Huang, J., Kelly, C.P., Bakirtzi, K. et al. Clostridium difficile toxins induce VEGF-A and vascular permeability to promote disease pathogenesis. Nat Microbiol 4, 269–279 (2019). https://doi.org/10.1038/s41564-018-0300-x
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DOI: https://doi.org/10.1038/s41564-018-0300-x
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