title: Helicobacter pylori heat-shock protein 60 induces inflammatory responses through the Toll-like receptor-triggered pathway in cultured human gastric epithelial cells.
abstract: Contact between Helicobacter pylori and gastric epithelial cells results in activation of NF-kappaB followed by secretion of interleukin (IL)-8. However, host-cell receptor(s) and their ligands involved in H. pylori-related IL-8 production have yet to be fully defined. In this study, the interaction between Toll-like receptors (TLRs), which are host receptors for pathogens involved in the innate immune response, and heat-shock protein (HSP) 60, an immune-potent antigen of H. pylori, was examined during H. pylori-induced IL-8 secretion in vitro. Recombinant H. pylori HSP60 (rHpHSP60) was prepared and added to cultured KATO III human gastric epithelial cells with or without pre-incubation with mouse monoclonal anti-TLR2 or anti-TLR4 antibodies. IL-8 mRNA expression and IL-8 protein release were analysed by Northern blotting and immunoassay. Involvement of NF-kappaB activation was analysed immunocytochemically by anti-NF-kappaB p65 antibody and ammonium pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-kappaB-mediated transcriptional activation. rHpHSP60 induced IL-8 mRNA expression and IL-8 secretion in a dose-dependent manner in KATO III cells. Anti-TLR2 antibody inhibited rHpHSP60-induced IL-8 secretion by 75 %, and anti-TLR4 antibody inhibited it by 30 %. rHpHSP60 induced nuclear translocation of NF-kappaB p65, which was inhibited by pretreatment with anti-TLR2 antibody. Treatment with PDTC significantly decreased the secretion of IL-8 induced by rHpHSP60. These findings suggest that H. pylori HSP60 activates NF-kappaB and induces IL-8 production through TLR-triggered pathways in gastric epithelial cells. Thus, it is possible that H. pylori HSP60 and TLR interaction in host cells contributes to the development of gastric inflammation caused by H. pylori infection.
authors.affiliation: Department of Medicine and Medical Science (Medicine 1), Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Ryuta
authors.initials: R
authors.lastname: Takenaka
authors.forename: Kenji
authors.initials: K
authors.lastname: Yokota
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Ayada
authors.forename: Motowo
authors.initials: M
authors.lastname: Mizuno
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Fujinami
authors.forename: Song-Nan
authors.initials: SN
authors.lastname: Lin
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Toyokawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Okada
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shiratori
authors.forename: Keiji
authors.initials: K
authors.lastname: Oguma
mesh_terms: D000818:Animals; D002460:Cell Line; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000382:microbiology*; D005753:Gastric Mucosa / Q000166:cytology / Q000382:microbiology*; D016480:Helicobacter pylori / Q000276:immunology / Q000472:pathogenicity*; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000209:etiology; D016209:Interleukin-8 / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors
chemical_list: D018834:Chaperonin 60; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D011956:Receptors, Cell Surface; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors