title: Salmonella enterica serovar Typhimurium regulates intercellular junction proteins and facilitates transepithelial neutrophil and bacterial passage.
abstract: The establishment of tight junctions (TJ) between columnar epithelial cells defines the functional barrier, which enteroinvasive pathogens have to overcome. Salmonella enterica serovar Typhimurium (S. typhimurium) directly invades intestinal epithelial cells but it is not well understood how the pathogen is able to overcome the intestinal barrier and gains access to the circulation. Therefore, we sought to determine whether infection with S. typhimurium could regulate the molecular composition of the TJ and, if so, whether these modifications would influence bacterial translocation and polymorphonuclear leukocyte (PMN) movement across model intestinal epithelium. We found that infection of a model intestinal epithelium with S. typhimurium over 2 h resulted in an approximately 80% loss of transepithelial electrical resistance. Western blot analysis of epithelial cell lysates demonstrated that S. typhimurium regulated the distribution of the TJ complex proteins claudin-1, zonula occludens (ZO)-2, and E-cadherin in Triton X-100-soluble and insoluble fractions. In addition, S. typhimurium was specifically able to dephosphorylate occludin and degrade ZO-1. This TJ alteration in the epithelial monolayer resulted in 10-fold increase in bacterial translocation and a 75% increase in N-formylmethionin-leucyl-phenyalanine-induced PMN transepithelial migration. Our data demonstrate that infection with S. typhimurium is associated with the rapid targeting of the tight junctional complex and loss of barrier function. This results in enhanced bacterial translocation and initiation of PMN migration across the intestinal barrier. Therefore, the ability to regulate the molecular composition of TJs facilitates the pathogenicity of S. typhimurium by aiding its uptake and distribution within the host.
authors.affiliation: Mucosal Immunology Laboratories, Massachusetts General Hospital, 114 16th Street, Charlestown, MA 02129, USA.
authors.forename: Henrik
authors.initials: H
authors.lastname: Köhler
authors.forename: Takanori
authors.initials: T
authors.lastname: Sakaguchi
authors.forename: Bryan P
authors.initials: BP
authors.lastname: Hurley
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Kase
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Kase
authors.forename: Hans-Christian
authors.initials: HC
authors.lastname: Reinecker
authors.forename: Beth A
authors.initials: BA
authors.lastname: McCormick
mesh_terms: D015820:Cadherins / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D062445:Claudin-1; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology*; D062793:Occludin; D010750:Phosphoproteins / Q000378:metabolism; D012486:Salmonella typhimurium / Q000378:metabolism* / Q000472:pathogenicity; D019108:Tight Junctions / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein
chemical_list: C568108:CLDN1 protein, human; D015820:Cadherins; D062445:Claudin-1; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein