Contact between and gastric epithelial cells results in activation of NF-B followed by secretion of interleukin (IL)-8. However, host-cell receptor(s) and their ligands involved in -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 , was examined during -induced IL-8 secretion . Recombinant 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-B activation was analysed immunocytochemically by anti-NF-B p65 antibody and ammonium pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-B-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-B 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 HSP60 activates NF-B and induces IL-8 production through TLR-triggered pathways in gastric epithelial cells. Thus, it is possible that HSP60 and TLR interaction in host cells contributes to the development of gastric inflammation caused by infection.


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