1887

Microbiology Society logo

Volume 157, Issue 8

Phosphorylation of the epidermal growth factor receptor (EGFR) is essential for interleukin-8 release from intestinal epithelial cells in response to challenge with O157 : H7 flagellin Free

Abstract

Enterohaemorrhagic O157 : H7 is a major foodborne and environmental pathogen responsible for both sporadic cases and outbreaks of food poisoning, which can lead to serious sequelae, such as haemolytic uraemic syndrome. The structural subunit of O157 : H7 flagella is flagellin, which is both the antigenic determinant of the H7 serotype, an important factor in colonization, and an immunomodulatory protein that has been determined to be a major pro-inflammatory component through the instigation of host cell signalling pathways. Flagellin has highly conserved N- and C-terminal regions that are recognized by the host cell pattern recognition receptor Toll-like receptor (TLR) 5. Activation of this receptor triggers cell signalling cascades, which are known to activate host cell kinases and transcription factors that respond with the production of inflammatory mediators such as the chemokine interleukin-8 (IL-8), although the exact components of this pathway are not yet fully characterized. We demonstrate that O157 : H7-derived flagellin induces rapid phosphorylation of the epidermal growth factor receptor (EGFR), as an early event in intestinal epithelial cell signalling, and that this is required for the release of the pro-inflammatory cytokine IL-8.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Funding
This study was supported by the:
  • Biotechnology and Biological Sciences Research Council
  • BBSRC (Award BB/C508485/1)
  • Rural and Environment Research and Analysis Directorate
  • RERAD
  • RERAD (Award 22050)
© 2011 SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.047670-0
2011-08-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/157/8/2339.html?itemId=/content/journal/micro/10.1099/mic.0.047670-0&mimeType=html&fmt=ahah

References

  1. Allen-Vercoe E., Dibb-Fuller M., Thorns C. J., Woodward M. J. ( 1997). SEF17 fimbriae are essential for the convoluted colonial morphology of Salmonella enteritidis . FEMS Microbiol Lett 153:33–42 [View Article][PubMed]
     [Google Scholar]
  2. Ballard D. W., Walker W. H., Doerre S., Sista P., Molitor J. A., Dixon E. P., Peffer N. J., Hannink M., Greene W. C. ( 1990). The v-rel oncogene encodes a kappa B enhancer binding protein that inhibits NF-kappa B function. Cell 63:803–814 [CrossRef]
     [Google Scholar]
  3. Basu S., Pathak S. K., Chatterjee G., Pathak S., Basu J., Kundu M. ( 2008). Helicobacter pylori protein HP0175 transactivates epidermal growth factor receptor through TLR4 in gastric epithelial cells. J Biol Chem 283:32369–32376 [View Article][PubMed]
     [Google Scholar]
  4. Berin M. C., Darfeuille-Michaud A., Egan L. J., Miyamoto Y., Kagnoff M. F. ( 2002). Role of EHEC O157 : H7 virulence factors in the activation of intestinal epithelial cell NF-κB and MAP kinase pathways and the upregulated expression of interleukin 8. Cell Microbiol 4:635–648 [View Article][PubMed]
     [Google Scholar]
  5. Best A., La Ragione R. M., Sayers A. R., Woodward M. J. ( 2005). Role for flagella but not intimin in the persistent infection of the gastrointestinal tissues of specific-pathogen-free chicks by Shiga toxin-negative Escherichia coli O157 : H7. Infect Immun 73:1836–1846 [View Article][PubMed]
     [Google Scholar]
  6. Boots A. W., Hristova M., Kasahara D. I., Haenen G. R., Bast A., van der Vliet A. ( 2009). ATP-mediated activation of the NADPH oxidase DUOX1 mediates airway epithelial responses to bacterial stimuli. J Biol Chem 284:17858–17867 [View Article][PubMed]
     [Google Scholar]
  7. Cario E., Gerken G., Podolsky D. K. ( 2004). Toll-like receptor 2 enhances ZO-1-associated intestinal epithelial barrier integrity via protein kinase C. Gastroenterology 127:224–238 [View Article][PubMed]
     [Google Scholar]
  8. Ding S. Z., Cho C. H., Lam S. K. ( 1997). Helicobacter pylori induces interleukin-8 expression in endothelial cells and the signal pathway is protein tyrosine kinase dependent. Biochem Biophys Res Commun 240:561–565 [View Article][PubMed]
     [Google Scholar]
  9. Dobbin H. S., Hovde C. J., Williams C. J., Minnich S. A. ( 2006). The Escherichia coli O157 flagellar regulatory gene flhC and not the flagellin gene fliC impacts colonization of cattle. Infect Immun 74:2894–2905 [View Article][PubMed]
     [Google Scholar]
  10. Egan L. J., de Lecea A., Lehrman E. D., Myhre G. M., Eckmann L., Kagnoff M. F. ( 2003). Nuclear factor-kappa B activation promotes restitution of wounded intestinal epithelial monolayers. Am J Physiol Cell Physiol 285:C1028–C1035[PubMed] [CrossRef]
     [Google Scholar]
  11. Erdem A. L., Avelino F., Xicohtencatl-Cortes J., Girón J. A. ( 2007). Host protein binding and adhesive properties of H6 and H7 flagella of attaching and effacing Escherichia coli . J Bacteriol 189:7426–7435 [View Article][PubMed]
     [Google Scholar]
  12. Finzi L., Shao M. X., Paye F., Housset C., Nadel J. A. ( 2009). Lipopolysaccharide initiates a positive feedback of epidermal growth factor receptor signaling by prostaglandin E2 in human biliary carcinoma cells. J Immunol 182:2269–2276 [View Article][PubMed]
     [Google Scholar]
  13. Franchi L., Amer A., Body-Malapel M., Kanneganti T. D., Ozören N., Jagirdar R., Inohara N., Vandenabeele P., Bertin J. et al. ( 2006). Cytosolic flagellin requires Ipaf for activation of caspase-1 and interleukin 1β in Salmonella-infected macrophages. Nat Immunol 7:576–582 [View Article][PubMed]
     [Google Scholar]
  14. Galán J. E., Pace J., Hayman M. J. ( 1992). Involvement of the epidermal growth factor receptor in the invasion of cultured mammalian cells by Salmonella typhimurium . Nature 357:588–589 [View Article][PubMed]
     [Google Scholar]
  15. Gao N., Kumar A., Jyot J., Yu F. S. ( 2010). Flagellin-induced corneal antimicrobial peptide production and wound repair involve a novel NF-κB-independent and EGFR-dependent pathway. PLoS ONE 5:e9351 [View Article][PubMed]
     [Google Scholar]
  16. Gazit A., Osherov N., Gilon C., Levitzki A. ( 1996). Tyrphostins. 6. Dimeric benzylidenemalononitrile tyrphostins: potent inhibitors of EGF receptor tyrosine kinase in vitro . J Med Chem 39:4905–4911 [View Article][PubMed]
     [Google Scholar]
  17. Ghosh S., Gifford A. M., Riviere L. R., Tempst P., Nolan G. P., Baltimore D. ( 1990). Cloning of the p50 DNA binding subunit of NF-kappa B: homology to rel and dorsal. Cell 62:1019–1029 [CrossRef]
     [Google Scholar]
  18. Girón J. A., Torres A. G., Freer E., Kaper J. B. ( 2002). The flagella of enteropathogenic Escherichia coli mediate adherence to epithelial cells. Mol Microbiol 44:361–379 [CrossRef]
     [Google Scholar]
  19. Gong Y., Zhao X. ( 2003). Shc-dependent pathway is redundant but dominant in MAPK cascade activation by EGF receptors: a modeling inference. FEBS Lett 554:467–472 [View Article][PubMed]
     [Google Scholar]
  20. Grif K., Orth D., Lederer I., Berghold C., Roedl S., Mache C. J., Dierich M. P., Würzner R. ( 2005). Importance of environmental transmission in cases of EHEC O157 causing hemolytic uremic syndrome. Eur J Clin Microbiol Infect Dis 24:268–271 [View Article][PubMed]
     [Google Scholar]
  21. Hayashi F., Smith K. D., Ozinsky A., Hawn T. R., Yi E. C., Goodlett D. R., Eng J. K., Akira S., Underhill D. M., Aderem A. ( 2001). The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410:1099–1103 [View Article][PubMed]
     [Google Scholar]
  22. Ibrahim G. F., Fleet G. H., Lyons M. J., Walker R. A. ( 1985). Method for the isolation of highly purified Salmonella flagellins. J Clin Microbiol 22:1040–1044[PubMed]
     [Google Scholar]
  23. Ivison S. M., Khan M. A., Graham N. R., Bernales C. Q., Kaleem A., Tirling C. O., Cherkasov A., Steiner T. S. ( 2007). A phosphorylation site in the Toll-like receptor 5 TIR domain is required for inflammatory signalling in response to flagellin. Biochem Biophys Res Commun 352:936–941 [View Article][PubMed]
     [Google Scholar]
  24. Iyoda S., Koizumi N., Satou H., Lu Y., Saitoh T., Ohnishi M., Watanabe H. ( 2006). The GrlR-GrlA regulatory system coordinately controls the expression of flagellar and LEE-encoded type III protein secretion systems in enterohemorrhagic Escherichia coli . J Bacteriol 188:5682–5692 [View Article][PubMed]
     [Google Scholar]
  25. Jansson M., Philipson L., Vennström B. ( 1983). Isolation and characterization of multiple human genes homologous to the oncogenes of avian erythroblastosis virus. EMBO J 2:561–565[PubMed]
     [Google Scholar]
  26. Jefferies C. A., Doyle S., Brunner C., Dunne A., Brint E., Wietek C., Walch E., Wirth T., O’Neill L. A. ( 2003). Bruton’s tyrosine kinase is a Toll/interleukin-1 receptor domain-binding protein that participates in nuclear factor κB activation by Toll-like receptor 4. J Biol Chem 278:26258–26264 [View Article][PubMed]
     [Google Scholar]
  27. Kawahara T., Kuwano Y., Teshima-Kondo S., Takeya R., Sumimoto H., Kishi K., Tsunawaki S., Hirayama T., Rokutan K. ( 2004). Role of nicotinamide adenine dinucleotide phosphate oxidase 1 in oxidative burst response to Toll-like receptor 5 signaling in large intestinal epithelial cells. J Immunol 172:3051–3058[PubMed] [CrossRef]
     [Google Scholar]
  28. Khan M. A., Bouzari S., Ma C., Rosenberger C. M., Bergstrom K. S., Gibson D. L., Steiner T. S., Vallance B. A. ( 2008). Flagellin-dependent and -independent inflammatory responses following infection by enteropathogenic Escherichia coli and Citrobacter rodentium . Infect Immun 76:1410–1422 [View Article][PubMed]
     [Google Scholar]
  29. Kieran M., Blank V., Logeat F., Vandekerckhove J., Lottspeich F., Le Bail O., Urban M. B., Kourilsky P., Baeuerle P. A., Israël A. ( 1990). The DNA binding subunit of NF-kappa B is identical to factor KBF1 and homologous to the rel oncogene product. Cell 62:1007–1018 [CrossRef]
     [Google Scholar]
  30. Kim J. G., Lee S. J., Kagnoff M. F. ( 2004). Nod1 is an essential signal transducer in intestinal epithelial cells infected with bacteria that avoid recognition by Toll-like receptors. Infect Immun 72:1487–1495 [View Article][PubMed]
     [Google Scholar]
  31. Koff J. L., Shao M. X., Kim S., Ueki I. F., Nadel J. A. ( 2006). Pseudomonas lipopolysaccharide accelerates wound repair via activation of a novel epithelial cell signaling cascade. J Immunol 177:8693–8700[PubMed] [CrossRef]
     [Google Scholar]
  32. Koff J. L., Shao M. X., Ueki I. F., Nadel J. A. ( 2008). Multiple TLRs activate EGFR via a signaling cascade to produce innate immune responses in airway epithelium. Am J Physiol Lung Cell Mol Physiol 294:L1068–L1075 [View Article][PubMed]
     [Google Scholar]
  33. Kogut M. H., Genovese K. J., He H., Kaiser P. ( 2008). Flagellin and lipopolysaccharide up-regulation of IL-6 and CXCLi2 gene expression in chicken heterophils is mediated by ERK1/2-dependent activation of AP-1 and NF-κB signaling pathways. Innate Immun 14:213–222 [View Article][PubMed]
     [Google Scholar]
  34. La Ragione R. M., Sayers A. R., Woodward M. J. ( 2000). The role of fimbriae and flagella in the colonization, invasion and persistence of Escherichia coli O78 : K80 in the day-old-chick model. Epidemiol Infect 124:351–363 [View Article][PubMed]
     [Google Scholar]
  35. Levitzki A. ( 1992). Tyrphostins: tyrosine kinase blockers as novel antiproliferative agents and dissectors of signal transduction. FASEB J 6:3275–3282[PubMed]
     [Google Scholar]
  36. Levitzki A., Gazit A. ( 1995). Tyrosine kinase inhibition: an approach to drug development. Science 267:1782–1788 [View Article][PubMed]
     [Google Scholar]
  37. Lipson K. E., Pang L., Huber L. J., Chen H., Tsai J. M., Hirth P., Gazit A., Levitzki A., McMahon G. ( 1998). Inhibition of platelet-derived growth factor and epidermal growth factor receptor signaling events after treatment of cells with specific synthetic inhibitors of tyrosine kinase phosphorylation. J Pharmacol Exp Ther 285:844–852[PubMed]
     [Google Scholar]
  38. Mahajan A., Currie C. G., Mackie S., Tree J., McAteer S., McKendrick I., McNeilly T. N., Roe A., La Ragione R. M. et al. ( 2009). An investigation of the expression and adhesin function of H7 flagella in the interaction of Escherichia coli O157 : H7 with bovine intestinal epithelium. Cell Microbiol 11:121–137 [View Article][PubMed]
     [Google Scholar]
  39. McNeilly T. N., Naylor S. W., Mahajan A., Mitchell M. C., McAteer S., Deane D., Smith D. G., Low J. C., Gally D. L., Huntley J. F. ( 2008). Escherichia coli O157 : H7 colonization in cattle following systemic and mucosal immunization with purified H7 flagellin. Infect Immun 76:2594–2602 [View Article][PubMed]
     [Google Scholar]
  40. McNeilly T. N., Mitchell M. C., Nisbet A. J., McAteer S., Erridge C., Inglis N. F., Smith D. G. E., Low J. C., Gally D. L., Huntley J. F. ( 2010). IgA and IgG antibody responses following systemic immunization of cattle with native H7 flagellin differ in epitope recognition and capacity to neutralise TLR5 signalling. Vaccine 28:1412–1421 [View Article][PubMed]
     [Google Scholar]
  41. Miao E. A., Alpuche-Aranda C. M., Dors M., Clark A. E., Bader M. W., Miller S. I., Aderem A. ( 2006). Cytoplasmic flagellin activates caspase-1 and secretion of interleukin 1β via Ipaf. Nat Immunol 7:569–575 [CrossRef]
     [Google Scholar]
  42. Miyamoto Y., Iimura M., Kaper J. B., Torres A. G., Kagnoff M. F. ( 2006). Role of Shiga toxin versus H7 flagellin in enterohaemorrhagic Escherichia coli signalling of human colon epithelium in vivo. Cell Microbiol 8:869–879 [View Article][PubMed]
     [Google Scholar]
  43. Naik S., Kelly E. J., Meijer L., Pettersson S., Sanderson I. R. ( 2001). Absence of Toll-like receptor 4 explains endotoxin hyporesponsiveness in human intestinal epithelium. J Pediatr Gastroenterol Nutr 32:449–453 [View Article][PubMed]
     [Google Scholar]
  44. Nataro J. P., Kaper J. B. ( 1998). Diarrheagenic Escherichia coli . Clin Microbiol Rev 11:142–201[PubMed]
     [Google Scholar]
  45. Ostroff S. M., Griffin P. M., Tauxe R. V., Shipman L. D., Greene K. D., Wells J. G., Lewis J. H., Blake P. A., Kobayashi J. M. ( 1990). A statewide outbreak of Escherichia coli O157 : H7 infections in Washington State. Am J Epidemiol 132:239–247[PubMed]
     [Google Scholar]
  46. Raimondi F., Santoro P., Barone M. V., Pappacoda S., Barretta M. L., Nanayakkara M., Apicella C., Capasso L., Paludetto R. ( 2008). Bile acids modulate tight junction structure and barrier function of Caco-2 monolayers via EGFR activation. Am J Gastrointest Liver Physiol 294:G906–G913 [CrossRef]
     [Google Scholar]
  47. Rajagopal S., Huang S., Moskal T. L., Lee B. N., el-Naggar A. K., Chakrabarty S. ( 1995). Epidermal growth factor expression in human colon and colon carcinomas: anti-sense epidermal growth factor receptor RNA down-regulates the proliferation of human colon cancer cells. Int J Cancer 62:661–667 [View Article][PubMed]
     [Google Scholar]
  48. Roxas J. L., Koutsouris A., Viswanathan V. K. ( 2007). Enteropathogenic Escherichia coli-induced epidermal growth factor receptor activation contributes to physiological alterations in intestinal epithelial cells. Infect Immun 75:2316–2324 [View Article][PubMed]
     [Google Scholar]
  49. Saha R. N., Jana M., Pahan K. ( 2007). MAPK p38 regulates transcriptional activity of NF-κB in primary human astrocytes via acetylation of p65. J Immunol 179:7101–7109[PubMed] [CrossRef]
     [Google Scholar]
  50. Shaykhiev R., Behr J., Bals R. ( 2008). Microbial patterns signalling via Toll-like receptors 2 and 5 contribute to epithelial repair, growth and survival. PLoS ONE 3:e1393 [View Article]
     [Google Scholar]
  51. Sherman P., Soni R., Yeger H. ( 1988). Characterization of flagella purified from enterohemorrhagic, vero-cytotoxin-producing Escherichia coli serotype O157 : H7. J Clin Microbiol 26:1367–1372[PubMed]
     [Google Scholar]
  52. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. ( 1985). Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85 [View Article][PubMed]
     [Google Scholar]
  53. Tjabringa G. S., Aarbiou J., Ninaber D. K., Drijfhout J. W., Sørensen O. E., Borregaard N., Rabe K. F., Hiemstra P. S. ( 2003). The antimicrobial peptide LL-37 activates innate immunity at the airway epithelial surface by transactivation of the epidermal growth factor receptor. J Immunol 171:6690–6696[PubMed] [CrossRef]
     [Google Scholar]
  54. Tong W. M., Ellinger A., Sheinin Y., Cross H. S. ( 1998). Epidermal growth factor receptor expression in primary cultured human colorectal carcinoma cells. Br J Cancer 77:1792–1798 [View Article][PubMed]
     [Google Scholar]
  55. Vijay-Kumar M., Wu H., Jones R., Grant G., Babbin B., King T. P., Kelly D., Gewirtz A. T., Neish A. S. ( 2006). Flagellin suppresses epithelial apoptosis and limits disease during enteric infection. Am J Pathol 169:1686–1700 [View Article][PubMed]
     [Google Scholar]
  56. Vijay-Kumar M., Wu H., Aitken J., Kolachala V. L., Neish A. S., Sitaraman S. V., Gewirtz A. T. ( 2007a). Activation of Toll-like receptor 3 protects against DSS-induced acute colitis. Inflamm Bowel Dis 13:856–864 [View Article][PubMed]
     [Google Scholar]
  57. Vijay-Kumar M., Sanders C. J., Taylor R. T., Kumar A., Aitken J. D., Sitaraman S. V., Neish A. S., Uematsu S., Akira S. et al. ( 2007b). Deletion of TLR5 results in spontaneous colitis in mice. J Clin Invest 117:3909–3921[PubMed]
     [Google Scholar]
  58. Vijay-Kumar M., Aitken J. D., Sanders C. J., Frias A., Sloane V. M., Xu J., Neish A. S., Rojas M., Gewirtz A. T. ( 2008). Flagellin treatment protects against chemicals, bacteria, viruses, and radiation. J Immunol 180:8280–8285[PubMed] [CrossRef]
     [Google Scholar]
  59. Yu Y., Zeng H., Lyons S., Carlson A., Merlin D., Neish A. S., Gewirtz A. T. ( 2003). TLR5-mediated activation of p38 MAPK regulates epithelial IL-8 expression via posttranscriptional mechanism. Am J Physiol Gastrointest Liver Physiol 285:G282–G290[PubMed] [CrossRef]
     [Google Scholar]
  60. Zhang Y., Wolf-Yadlin A., Ross P. L., Pappin D. J., Rush J., Lauffenburger D. A., White F. M. ( 2005). Time-resolved mass spectrometry of tyrosine phosphorylation sites in the epidermal growth factor receptor signaling network reveals dynamic modules. Mol Cell Proteomics 4:1240–1250 [CrossRef]
     [Google Scholar]
  61. Zhou X., Girón J. A., Torres A. G., Crawford J. A., Negrete E., Vogel S. N., Kaper J. B. ( 2003). Flagellin of enteropathogenic Escherichia coli stimulates interleukin-8 production in T84 cells. Infect Immun 71:2120–2129 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.047670-0
Loading
Phosphorylation of the epidermal growth factor receptor (EGFR) is essential for interleukin-8 release from intestinal epithelial cells in response to challenge with Escherichia coli O157 : H7 flagellin
Microbiology 157, 2339 (2011); https://doi.org/10.1099/mic.0.047670-0
/content/journal/micro/10.1099/mic.0.047670-0
/content/journal/micro/10.1099/mic.0.047670-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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