1887

Abstract

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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27527-0
2004-12-01
2019-11-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/12/mic1503913.html?itemId=/content/journal/micro/10.1099/mic.0.27527-0&mimeType=html&fmt=ahah

References

  1. Aderem, A. & Ulevitch, R. J. ( 2000; ). Toll-like receptors in the induction of the innate immune response. Nature 406, 782–787.[CrossRef]
    [Google Scholar]
  2. Akira, S. & Hemmi, H. ( 2003; ). Recognition of pathogen-associated molecular patterns by TLR family. Immunol Lett 85, 85–95.[CrossRef]
    [Google Scholar]
  3. Bhattacharyya, A., Pathak, S., Datta, S., Chattopadhyay, S., Basu, J. & Kundu, M. ( 2002; ). Mitogen-activated protein kinases and nuclear factor-kappaB regulate Helicobacter pylori-mediated interleukin-8 release from macrophages. Biochem J 368, 121–129.[CrossRef]
    [Google Scholar]
  4. Blaser, M. J. ( 1990; ). Helicobacter pylori and the pathogenesis of gastroduodenal inflammation. J Infect Dis 161, 626–633.[CrossRef]
    [Google Scholar]
  5. Covacci, A., Telford, J. L., Del Giudice, G., Parsonnet, J. & Rappuoli, R. ( 1999; ). Helicobacter pylori virulence and genetic geography. Science 284, 1328–1333.[CrossRef]
    [Google Scholar]
  6. Crabtree, J. E., Wyatt, J. I., Trejdosiewicz, L. K., Peichl, P., Nichols, P. H., Ramsay, N., Primrose, J. N. & Lindley, I. J. ( 1994; ). Interleukin-8 expression in Helicobacter pylori infected, normal, and neoplastic gastroduodenal mucosa. J Clin Pathol 47, 61–66.[CrossRef]
    [Google Scholar]
  7. Crabtree, J. E., Covacci, A., Farmery, S. M., Xiang, Z., Tompkins, D. S., Perry, S., Lindley, I. J. & Rappuoli, R. ( 1995a; ). Helicobacter pylori induced interleukin-8 expression in gastric epithelial cells is associated with CagA positive phenotype. J Clin Pathol 48, 41–45.[CrossRef]
    [Google Scholar]
  8. Crabtree, J. E., Xiang, Z., Lindley, I. J., Tompkins, D. S., Rappuoli, R. & Covacci, A. ( 1995b; ). Induction of interleukin-8 secretion from gastric epithelial cells by a cagA negative isogenic mutant of Helicobacter pylori. J Clin Pathol 48, 967–969.[CrossRef]
    [Google Scholar]
  9. Cunningham, M. D., Shapiro, R. A., Seachord, C., Ratcliffe, K., Cassiano, L. & Darveau, R. P. ( 2000; ). CD14 employs hydrophilic regions to ‘capture’ lipopolysaccharides. J Immunol 164, 3255–3263.[CrossRef]
    [Google Scholar]
  10. Ellis, R. J. ( 1990; ). The molecular chaperone concept. Semin Cell Biol 1, 1–9.
    [Google Scholar]
  11. Gao, B. & Tsan, M. F. ( 2003; ). Recombinant human heat shock protein 60 does not induce the release of tumor necrosis factor alpha from murine macrophages. J Biol Chem 278, 22523–22529.[CrossRef]
    [Google Scholar]
  12. Gobert, A. P., Bambou, J. C., Werts, C., Balloy, V., Chignard, M., Moran, A. P. & Ferrero, R. L. ( 2004; ). Helicobacter pylori heat shock protein 60 mediates interleukin-6 production by macrophages via a toll-like receptor (TLR)-2-, TLR-4-, and myeloid differentiation factor 88-independent mechanism. J Biol Chem 279, 245–250.[CrossRef]
    [Google Scholar]
  13. Hayashi, S., Sugiyama, T., Yokota, K., Isogai, H., Isogai, E., Oguma, K., Asaka, M., Fujii, N. & Hirai, Y. ( 1998; ). Analysis of immunoglobulin A antibodies to Helicobacter pylori in serum and gastric juice in relation to mucosal inflammation. Clin Diagn Lab Immunol 5, 617–621.
    [Google Scholar]
  14. Huesca, M., Borgia, S., Hoffman, P. & Lingwood, C. A. ( 1996; ). Acidic pH changes receptor binding specificity of Helicobacter pylori: a binary adhesion model in which surface heat shock (stress) proteins mediate sulfatide recognition in gastric colonization. Infect Immun 64, 2643–2648.
    [Google Scholar]
  15. Ishii, E., Yokota, K., Sugiyama, T. & 7 other authors ( 2001; ). Immunoglobulin G1 antibody response to Helicobacter pylori heat shock protein 60 is closely associated with low-grade gastric mucosa-associated lymphoid tissue lymphoma. Clin Diagn Lab Immunol 8, 1056–1059.
    [Google Scholar]
  16. Kawahara, Y., Yokota, K., Mizuno, M. & 7 other authors ( 1999; ). Antibodies to human gastric epithelial cells and heat shock protein 60 in Helicobacter pylori positive mucosa associated lymphoid tissue lymphoma. Gut 45, 20–23.[CrossRef]
    [Google Scholar]
  17. Kawahara, T., Teshima, S., Oka, A., Sugiyama, T., Kishi, K. & Rokutan, K. ( 2001; ). Type I Helicobacter pylori lipopolysaccharide stimulates toll-like receptor 4 and activates mitogen oxidase 1 in gastric pit cells. Infect Immun 69, 4382–4389.[CrossRef]
    [Google Scholar]
  18. Kawai, M., Nishikomori, R., Jung, E. Y., Tai, G., Yamanaka, C., Mayumi, M. & Heike, T. ( 1995; ). Pyrrolidine dithiocarbamate inhibits intercellular adhesion molecule-1 biosynthesis induced by cytokines in human fibroblasts. J Immunol 154, 2333–2341.
    [Google Scholar]
  19. Lee, S. K., Stack, A., Katzowitsch, E., Aizawa, S. I., Suerbaum, S. & Josenhans, C. ( 2003; ). Helicobacter pylori flagellins have very low intrinsic activity to stimulate human gastric epithelial cells via TLR5. Microbes Infect 5, 1345–1356.[CrossRef]
    [Google Scholar]
  20. Mahmut, N., Inoue, K., Fujinaga, Y. & 7 other authors ( 2002; ). Characterisation of monoclonal antibodies against haemagglutinin associated with Clostridium botulinum type C neurotoxin. J Med Microbiol 51, 286–294.
    [Google Scholar]
  21. Meyer-ter-Vehn, T., Covacci, A., Kist, M. & Pahl, H. L. ( 2000; ). Helicobacter pylori activates mitogen-activated protein kinase cascades and induces expression of the proto-oncogenes c-fos and c-jun. J Biol Chem 275, 16064–16072.[CrossRef]
    [Google Scholar]
  22. Ohashi, K., Burkart, V., Flohe, S. & Kolb, H. ( 2000; ). Cutting edge: heat shock protein 60 is a putative endogenous ligand of the toll-like receptor-4 complex. J Immunol 164, 558–561.[CrossRef]
    [Google Scholar]
  23. Parsonnet, J., Friedman, G. D., Vandersteen, D. P., Chang, Y., Vogelman, J. H., Orentreich, N. & Sibley, R. K. ( 1991; ). Helicobacter pylori infection and the risk of gastric carcinoma. N Engl J Med 325, 1127–1131.[CrossRef]
    [Google Scholar]
  24. Parsonnet, J., Hansen, S., Rodriguez, L., Gelb, A. B., Warnke, R. A., Jellum, E., Orentreich, N., Vogelman, J. H. & Friedman, G. D. ( 1994; ). Helicobacter pylori infection and gastric lymphoma. N Engl J Med 330, 1267–1271.[CrossRef]
    [Google Scholar]
  25. Rad, R., Gerhard, M., Lang, R., Schoniger, M., Rosch, T., Schepp, W., Becker, I., Wagner, H. & Prinz, C. ( 2002; ). The Helicobacter pylori blood group antigen-binding adhesin facilitates bacterial colonization and augments a nonspecific immune response. J Immunol 168, 3033–3041.[CrossRef]
    [Google Scholar]
  26. Sasu, S., LaVerda, D., Qureshi, N., Golenbock, D. T. & Beasley, D. ( 2001; ). Chlamydia pneumoniae and chlamydial heat shock protein 60 stimulate proliferation of human vascular smooth muscle cells via toll-like receptor 4 and p44/p42 mitogen-activated protein kinase activation. Circ Res 89, 244–250.[CrossRef]
    [Google Scholar]
  27. Sharma, S. A., Miller, G. G., Peek, R. J., Jr, Perez-Perez, G. & Blaser, M. J. ( 1997; ). T-cell, antibody, and cytokine responses to homologs of the 60-kilodalton heat shock protein in Helicobacter pylori infection. Clin Diagn Lab Immunol 4, 440–446.
    [Google Scholar]
  28. Sharma, S. A., Tummuru, M. K., Blaser, M. J. & Kerr, L. D. ( 1998; ). Activation of IL-8 gene expression by Helicobacter pylori is regulated by transcription factor nuclear factor-kappa B in gastric epithelial cells. J Immunol 160, 2401–2407.
    [Google Scholar]
  29. Smith, M. F., Jr, Mitchell, A., Li, G., Ding, S., Fitzmaurice, A. M., Ryan, K., Crowe, S. & Goldberg, J. B. ( 2003; ). Toll-like receptor (TLR) 2 and TLR5, but not TLR4, are required for Helicobacter pylori-induced NF-kappa B activation and chemokine expression by epithelial cells. J Biol Chem 278, 32552–32560.[CrossRef]
    [Google Scholar]
  30. Stokes, D. C., Shenep, J. L., Fishman, M., Hildner, W. K., Bysani, G. K. & Rufus, K. ( 1989; ). Polymyxin B prevents lipopolysaccharide-induced release of tumor necrosis factor-alpha from alveolar macrophages. J Infect Dis 160, 52–57.[CrossRef]
    [Google Scholar]
  31. Vabulas, R. M., Ahmad-Nejad, P., da Costa, C., Miethke, T., Kirschning, C. J., Hacker, H. & Wagner, H. ( 2001; ). Endocytosed HSP60s use toll-like receptor 2 (TLR2) and TLR4 to activate the toll/interleukin-1 receptor signaling pathway in innate immune cells. J Biol Chem 276, 31332–31339.[CrossRef]
    [Google Scholar]
  32. Yamaguchi, H., Osaki, T., Kurihara, N., Taguchi, H., Hanawa, T., Yamamoto, T. & Kamiya, S. ( 1997; ). Heat-shock protein 60 homologue of Helicobacter pylori is associated with adhesion of H. pylori to human gastric epithelial cells. J Med Microbiol 46, 825–831.[CrossRef]
    [Google Scholar]
  33. Yamaguchi, H., Osaki, T., Kurihara, N. Kitajima M., Kai, M., Takahashi, M., Taguchi, H. & Kamiya, S. ( 1999; ). Induction of secretion of interleukin-8 from human gastric epithelial cells by heat-shock protein 60 homologue of Helicobacter pylori. J Med Microbiol 48, 927–933.[CrossRef]
    [Google Scholar]
  34. Yamaoka, Y., Kikuchi, S., el-Zimaity, H. M., Gutierrez, O., Osato, M. S. & Graham, D. Y. ( 2002; ). Importance of Helicobacter pylori oipA in clinical presentation, gastric inflammation, and mucosal interleukin 8 production. Gastroenterology 123, 414–424.[CrossRef]
    [Google Scholar]
  35. Young, D. B. ( 1990; ). Chaperonins and the immune response. Semin Cell Biol 1, 27–35.
    [Google Scholar]
  36. Yunoki, N., Yokota, K., Mizuno, M. & 7 other authors ( 2000; ). Antibody to heat shock protein can be used for early serological monitoring of Helicobacter pylori eradication treatment. Clin Diagn Lab Immunol 7, 574–577.
    [Google Scholar]
  37. Ziegler-Heitbrock, H. W., Sternsdorf, T., Liese, J., Belohradsky, B., Weber, C., Wedel, A., Schreck, R., Bauerle, P. & Strobel, M. ( 1993; ). Pyrrolidine dithiocarbamate inhibits NF-kappa B mobilization and TNF production in human monocytes. J Immunol 151, 6986–6993.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27527-0
Loading
/content/journal/micro/10.1099/mic.0.27527-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error