title: Epithelial cells augment barrier function via activation of the Toll-like receptor 2/phosphatidylinositol 3-kinase pathway upon recognition of Salmonella enterica serovar Typhimurium curli fibrils in the gut.
abstract: Curli fibrils, the best-characterized functional bacterial amyloids, are an important component of enterobacterial biofilms. We have previously shown that curli fibrils are recognized by the Toll-like receptor 2 (TLR2)/TLR1 heterodimer complex. Utilizing polarized T-84 cells, an intestinal epithelial cell line derived from colon carcinoma grown on semipermeable tissue culture inserts, we determined that infection with a Salmonella enterica serovar Typhimurium csgBA mutant, which does not express curli, resulted in an increase in intestinal barrier permeability and an increase in bacterial translocation compared to infection with curliated wild-type S. Typhimurium. When the TLR2 downstream signaling molecule phosphatidylinositol 3-kinase (PI3K) was blocked using wortmannin or LY294002, the difference in disruption of the intestinal epithelium and bacterial translocation was no longer observed. Additionally, disruption of polarized T-84 cells treated basolaterally with the TLR5 ligand flagellin was prevented when the polarized cells were simultaneously treated with the synthetic TLR2/TLR1 ligand Pam(3)CSK(4) or with purified curli fibrils in the apical compartment. Similar to in vitro observations, C57BL/6 mice infected with the csgBA mutant suffered increased disruption of the intestinal epithelium and therefore greater dissemination of the bacteria to the mesenteric lymph nodes than mice infected with wild-type S. Typhimurium. The differences in disruption of the intestinal epithelium and bacterial dissemination in the mice infected with csgBA mutant or wild-type S. Typhimurium were not apparent in TLR2-deficient mice. Overall, these studies report for the first time that activation of the TLR2/PI3K pathway by microbial amyloids plays a critical role in regulating the intestinal epithelial barrier as well as monitoring bacterial translocation during infection.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, Temple University, Philadelphia, PA, USA.
authors.forename: Gertrude O
authors.initials: GO
authors.lastname: Oppong
authors.forename: Glenn J
authors.initials: GJ
authors.lastname: Rapsinski
authors.forename: Tiffanny N
authors.initials: TN
authors.lastname: Newman
authors.forename: Jessalyn H
authors.initials: JH
authors.lastname: Nishimori
authors.forename: Steven G
authors.initials: SG
authors.lastname: Biesecker
authors.forename: Çagla
authors.initials: Ç
authors.lastname: Tükel
mesh_terms: D000682:Amyloid / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000378:metabolism* / Q000382:microbiology; D005260:Female; D005408:Flagellin / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000378:metabolism / Q000382:microbiology; D006367:HeLa Cells; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000382:microbiology; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D010539:Permeability; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D019779:Salmonella enterica / Q000378:metabolism*; D015398:Signal Transduction; D051194:Toll-Like Receptor 1 / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000378:metabolism*
chemical_list: D000682:Amyloid; D001426:Bacterial Proteins; D016209:Interleukin-8; D008024:Ligands; C495342:TLR2 protein, human; D051194:Toll-Like Receptor 1; D051195:Toll-Like Receptor 2; D005408:Flagellin; C077481:Crl protein, Bacteria; D019869:Phosphatidylinositol 3-Kinases