Mucosal barriers segregate commensal microbes from the intestinal epithelia to maintain gut homeostasis. Ly6/Plaur domain-containing 8 (Lypd8), a highly glycosylated glycosylphosphatidylinositol-anchored protein selectively expressed on colonic enterocytes, promotes this segregation by inhibiting bacterial invasion of the inner mucus layer and colonic epithelia. However, it remains unclear whether Lypd8 prevents infection with enteric bacterial pathogens. Here, we demonstrate that Lypd8 strongly contributes to early-phase defense against Citrobacter rodentium, which causes colitis by inducing attachment and effacement (A/E) lesions on colonic epithelia. Lypd8 inhibits C. rodentium attachment to intestinal epithelial cells by binding to intimin, thereby suppressing the interaction between intimin and translocated intimin receptor. Lypd8 deficiency leads to rapid C. rodentium colonization in the colon, resulting in severe colitis with Th17-cell and neutrophil expansion in the lamina propria. This study identifies a novel function for Lypd8 against A/E bacteria and highlights the role of enterocytes as crucial players in innate immunity for protection against enteric bacterial pathogens.
Subscribe to Journal
Get full journal access for 1 year
only $92.33 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Collins, J. W. et al. Citrobacter rodentium: infection, inflammation and the microbiota. Nat. Rev. Microbiol. 12, 612–623 (2014).
Majowicz, S. E. et al. Global incidence of human Shiga toxin-producing Escherichia coli infections and deaths: a systematic review and knowledge synthesis. Foodborne Pathog. Dis. 11, 447–455 (2014).
Maynard, C. L., Elson, C. O., Hatton, R. D. & Weaver, C. T. Reciprocal interactions of the intestinal microbiota and immune system. Nature 489, 231–241 (2012).
Okumura, R. & Takeda, K. Roles of intestinal epithelial cells in the maintenance of gut homeostasis. Exp. Mol. Med. 49, e338 (2017).
Bergstrom, K. S. et al. Muc2 protects against lethal infectious colitis by disassociating pathogenic and commensal bacteria from the colonic mucosa. PLoS Pathog. 6, e1000902 (2010).
Goto, Y. et al. Innate lymphoid cells regulate intestinal epithelial cell glycosylation. Science 345, 1254009 (2014).
van Ampting, M. T. et al. Intestinally secreted C-type lectin Reg3b attenuates salmonellosis but not listeriosis in mice. Infect. Immun. 80, 1115–1120 (2012).
Wilson, C. L. et al. Regulation of intestinal alpha-defensin activation by the metalloproteinase matrilysin in innate host defense. Science 286, 113–117 (1999).
Okumura, R. et al. Lypd8 promotes the segregation of flagellated microbiota and colonic epithelia. Nature 532, 117–121 (2016).
Gaytan, M. O., Martinez-Santos, V. I., Soto, E. & Gonzalez-Pedrajo, B. Type three secretion system in attaching and effacing pathogens. Front. Cell. Infect. Microbiol. 6, 129 (2016).
Kenny, B. et al. Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91, 511–520 (1997).
Liu, H., Magoun, L., Luperchio, S., Schauer, D. B. & Leong, J. M. The Tir-binding region of enterohaemorrhagic Escherichia coli intimin is sufficient to trigger actin condensation after bacterial-induced host cell signalling. Mol. Microbiol. 34, 67–81 (1999).
Zheng, Y. et al. Interleukin-22 mediates early host defense against attaching and effacing bacterial pathogens. Nat. Med. 14, 282–289 (2008).
Backert, I. et al. STAT3 activation in Th17 and Th22 cells controls IL-22-mediated epithelial host defense during infectious colitis. J. Immunol. 193, 3779–3791 (2014).
Aychek, T. et al. IL-23-mediated mononuclear phagocyte crosstalk protects mice from Citrobacter rodentium-induced colon immunopathology. Nat. Commun. 6, 6525 (2015).
Atarashi, K. et al. Th17 cell induction by adhesion of microbes to intestinal epithelial cells. Cell 163, 367–380 (2015).
Sassone-Corsi, M. & Raffatellu, M. No vacancy: how beneficial microbes cooperate with immunity to provide colonization resistance to pathogens. J. Immunol. 194, 4081–4087 (2015).
McKee, M. L., Melton-Celsa, A. R., Moxley, R. A., Francis, D. H. & O'Brien, A. D. Enterohemorrhagic Escherichia coli O157:H7 requires intimin to colonize the gnotobiotic pig intestine and to adhere to HEp-2 cells. Infect. Immun. 63, 3739–3744 (1995).
Petty, N. K. et al. Citrobacter rodentium is an unstable pathogen showing evidence of significant genomic flux. PLoS Pathog. 7, e1002018 (2011).
Khan, M. A. et al. Flagellin-dependent and -independent inflammatory responses following infection by enteropathogenic Escherichia coli and Citrobacter rodentium. Infect. Immun. 76, 1410–1422 (2008).
Zhang, X. W. et al. A C-type lectin with an immunoglobulin-like domain promotes phagocytosis of hemocytes in crayfish Procambarus clarkii. Sci. Rep. 6, 29924 (2016).
Kelly, G. et al. Structure of the cell-adhesion fragment of intimin from enteropathogenic Escherichia coli. Nat. Struct. Biol. 6, 313–318 (1999).
Notti, R. Q. & Stebbins, C. E. The structure and function of type III secretion systems. Microbiol. Spectr. 4, https://doi.org/10.1128/microbiolspec.VMBF-0004-2015 (2016).
Batchelor, M. et al. Structural basis for recognition of the translocated intimin receptor (Tir) by intimin from enteropathogenic Escherichia coli. EMBO J. 19, 2452–2464 (2000).
Loonen, L. M. et al. REG3gamma-deficient mice have altered mucus distribution and increased mucosal inflammatory responses to the microbiota and enteric pathogens in the ileum. Mucosal Immunol. 7, 939–947 (2014).
Mellies, J. L. & Lorenzen, E. Enterohemorrhagic Escherichia coli virulence gene. Regul. Microbiol. Spectr. 2, EHEC-0004–EHEC-2013 (2014).
Mangan, P. R. et al. Transforming growth factor-beta induces development of the T(H)17 lineage. Nature 441, 231–234 (2006).
Lebeis, S. L., Bommarius, B., Parkos, C. A., Sherman, M. A. & Kalman, D. TLR signaling mediated by MyD88 is required for a protective innate immune response by neutrophils to Citrobacter rodentium. J. Immunol. 179, 566–577 (2007).
Giron, J. A., Torres, A. G., Freer, E. & Kaper, J. B. The flagella of enteropathogenic Escherichia coli mediate adherence to epithelial cells. Mol. Microbiol. 44, 361–379 (2002).
Gavin, R. et al. Lateral flagella of Aeromonas species are essential for epithelial cell adherence and biofilm formation. Mol. Microbiol. 43, 383–397 (2002).
Liu, H. et al. Point mutants of EHEC intimin that diminish Tir recognition and actin pedestal formation highlight a putative Tir binding pocket. Mol. Microbiol. 45, 1557–1573 (2002).
Yi, Y. et al. Crystal structure of EHEC intimin: insights into the complementarity between EPEC and EHEC. PLoS One 5, e15285 (2010).
We thank T. Kondo and Y. Magota for technical assistance, Y. Fujioka, T. Nakano, and K. Sano at Osaka Medical College for analysis of bacterial morphology, T. Sheen at Edanz Group for editing a draft of this paper, and C. Hidaka for secretarial assistance. The transmission electron microscopic study was supported by E. Oiki and N. Hayakawa at the Center for Medical Research and Education, Graduate School of Medicine, Osaka University. The surface plasmon resonance interaction analysis using Biacore was supported by Y. Ito at the Center for Medical Research and Education, Graduate School of Medicine, Osaka University. This study was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (A18H040280 and T18K151870), the Japan Agency for Medical Research and Development (JP18gm1010004), and the Terumo Foundation for Life Sciences and Arts.
The authors declare no competing interests.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Okumura, R., Kodama, T., Hsu, C. et al. Lypd8 inhibits attachment of pathogenic bacteria to colonic epithelia. Mucosal Immunol 13, 75–85 (2020). https://doi.org/10.1038/s41385-019-0219-4