title: Salmonella typhimurium transcytoses flagellin via an SPI2-mediated vesicular transport pathway.
abstract: Apical colonization of polarized epithelia by Salmonella typhimurium results in translocation of flagellin to the basolateral membrane domain, thus enabling activation of toll-like receptor 5 (TLR5)-mediated pro-inflammatory gene expression. Such flagellin transcytosis occurred without a change in epithelial permeability to 40 kDa FITC dextran, did not require bacterial motility and was independent of transepithelial movement of intact bacteria. Flagellin transcytosis was blocked at 20 degrees C, suggesting dependence on vesicular transport consistent with results from confocal microscopy that showed flagellin independent of bacteria inside epithelial cells. Furthermore, vesicles isolated from S. typhimurium-infected epithelia were highly enriched in flagellin. Flagellin transcytosis was dependent upon genes of Salmonella pathogenicity island (SPI)-2, which alter vesicular trafficking, but independent of SPI-1 that mediates bacterial invasion. Furthermore, such SPI-2 mutants were unable to mediate the localization of flagellin into intracellular vesicles consistent with flagellin transcytosis mediated by a S. typhimurium take-over of host vesicle trafficking pathways. As a result of their inability to transcytose flagellin, apical colonization by SPI-2 mutants induced substantially less epithelial IL-8 secretion than wild-type strains suggesting that such SPI-2 mediated transcytosis of flagellin plays a role in the pathogenesis of the mucosal inflammation characteristic of human Salmonellosis.
authors.affiliation: Epithelial Pathobiology Division, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA 30322, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Lyons
authors.forename: Lixin
authors.initials: L
authors.lastname: Wang
authors.forename: James E
authors.initials: JE
authors.lastname: Casanova
authors.forename: Shanthi V
authors.initials: SV
authors.lastname: Sitaraman
authors.forename: Didier
authors.initials: D
authors.lastname: Merlin
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Gewirtz
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D002460:Cell Line; D004285:Dogs; D004848:Epithelium / Q000378:metabolism / Q000382:microbiology*; D005408:Flagellin / Q000378:metabolism*; D044404:Genomic Islands; D007249:Inflammation; D016209:Interleukin-8 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009154:Mutation; D021381:Protein Transport; D011956:Receptors, Cell Surface / Q000378:metabolism; D012486:Salmonella typhimurium / Q000472:pathogenicity* / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D013696:Temperature; D013997:Time Factors; D051198:Toll-Like Receptor 5; D051193:Toll-Like Receptors
chemical_list: D001426:Bacterial Proteins; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; C440638:SPI-2 protein, Salmonella; D051198:Toll-Like Receptor 5; D051193:Toll-Like Receptors; D005408:Flagellin