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  • 1 Helmholtz Zentrum München, Neuherberg, Germany
  • 2 Ludwig Maximilians University, Munich, Germany
  • 3 Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, D-85764 Oberschleissheim, Germany

Interleukin-8 (IL-8) is a potent neutrophil-activating chemokine which triggers the infiltration and migration of neutrophils into areas of bacterial infection. Helicobacter pylori-infected patient studies as well as animal models have revealed that H. pylori type I strains carrying an intact cytotoxin-associated gene pathogenicity island (cag-PAI) with a functional type IV secretion system (T4SS) induce IL-8 expression and secretion in gastric mucosa. This gastric mucosal IL-8 expression correlates with severe histological changes due to H. pylori infection.

In the present study, we explored a new recognition pattern on the bacterial adhesion protein CagL inducing IL-8 expression in H. pylori-infected host cells. To analyze the secreted IL-8 concentration, we performed IL-8 enzyme-linked immunosorbent assay (ELISA). To investigate the H. pylori-induced IL-8 expression on the transcriptional level, we transiently transfected gastric epithelial cells (AGS) with a human IL-8 luciferase reporter construct.

The results of this study demonstrate that specifically the C-terminal coiled-coil region of the H. pylori CagL protein, a protein described to be located on the tip of the T4SS-pilus, is responsible for several in vitro observations: 1) H. pylori-induced IL-8 secretion via the transforming growth factor (TGF)-α activated epidermal growth factor-receptor (EGF-R) signaling pathway; 2) H. pylori-induced elongation of the cells, a typical CagA-induced phenotype; and 3) the bridging of the T4SS to its human target cells. This novel bacterial-host recognition sequence allows a new insight into how H. pylori induces the inflammatory response in gastric epithelial cells and facilitates the development of precancerous conditions.

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  • 1.

    Houghton J , Wang TC: Helicobacter pylori and gastric cancer: a new paradigm for inflammation-associated epithelial cancers. Gastroenterology 128(6), 15671578 (2005)

    • Search Google Scholar
    • Export Citation
  • 2.

    Lee A , Fox J, Hazell S: Pathogenicity of Helicobacter pylori: a perspective. Infect Immun 61(5), 16011610 (1993)

  • 3.

    El-Omar EM , Carrington M, Chow WH, McColl KE, Bream JH, Young HA Herrera J, Lissowska J, Yuan CC, Rothman N, Lanyon G, Martin M, Fraumeni JF Jr, Rabkin CS: Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature 404(6776), 398402 (2000)

    • Search Google Scholar
    • Export Citation
  • 4.

    Rieder G , Fischer W, Haas R: Interaction of Helicobacter pylori with host cells: function of secreted and translocated molecules. Curr Opin Microbiol 8(1), 6773 (2005)

    • Search Google Scholar
    • Export Citation
  • 5.

    Ogura K , Maeda S, Nakao M, Watanabe T, Tada M, Kyutoku T, et al.: Virulence factors of Helicobacter pylori responsible for gastric diseases in Mongolian gerbil. J Exp Med 192(11), 16011610 (2000)

    • Search Google Scholar
    • Export Citation
  • 6.

    Rieder G , Merchant JL, Haas R: Helicobacter pylori cagtype IV secretion system facilitates corpus colonization to induce precancerous conditions in Mongolian gerbils. Gastroenterology 128(5), 12291242 (2005)

    • Search Google Scholar
    • Export Citation
  • 7.

    Wiedemann T , Loell E, Mueller S, Stoeckelhuber M, Stolte M, Haas R, Rieder G: Helicobacter pylori cag-pathogenicity island-dependent early immunological response triggers later precancerous gastric changes in Mongolian gerbils. PLoS One 4(3), e4754 (2009)

    • Search Google Scholar
    • Export Citation
  • 8.

    Baggiolini M , Loetscher P, Moser B: Interleukin-8 and the chemokine family. Int, J Immunopharmacol 17(2), 103108 (1995)

  • 9.

    Crabtree JE , Lindley IJ: Mucosal interleukin-8 and Helicobacter pylori-associated gastroduodenal disease. Eur, J Gastroenterol Hepatol 6 Suppl 1, S3338 (1994)

    • Search Google Scholar
    • Export Citation
  • 10.

    Ando T , Kusugami K, Ohsuga M, Shinoda M, Sakakibara M, Saito H, Fukatsu A, Ichiyama S, Ohta M: Interleukin-8 activity correlates with histological severity in Helicobacter pylori-associated antral gastritis. Am, J Gastroenterol 91(6), 11501156 (1996)

    • Search Google Scholar
    • Export Citation
  • 11.

    Lee KE , Khoi PN, Xia Y, Park JS, Joo YE, Kim KK, Choi SY, Jung YD: Helicobacter pylori and interleukin-8 in gastric cancer. World, J Gastroenterol 19(45), 819281202 (2013)

    • Search Google Scholar
    • Export Citation
  • 12.

    Siddique I , Al-Qabandi A, Al-Ali J, Alazmi W, Memon A, Mustafa AS, Junaid TA: Association between Helicobacter pylori genotypes and severity of chronic gastritis, peptic ulcer disease and gastric mucosal interleukin-8 levels: evidence from a study in the Middle East. Gut Pathog 6(1), 41 (2014)

    • Search Google Scholar
    • Export Citation
  • 13.

    Chongruksut W , Limpakan Yamada S, Chakrabandhu B, Ruengorn C, Nanta S: Correlation of Helicobacter pylori and interleukin-8 mRNA expression in high risk gastric cancer population prediction. World, J Gastrointest Oncol 8(2), 215221 (2016)

    • Search Google Scholar
    • Export Citation
  • 14.

    Fischer W , Puls J, Buhrdorf R, Gebert B, Odenbreit S, Haas R: Systematic mutagenesis of the Helicobacter pylori cag pathogenicity island: essential genes for CagA translocation in host cells and induction of interleukin-8. Mol Microbiol 42(5), 13371348 (2001)

    • Search Google Scholar
    • Export Citation
  • 15.

    Barrozo RM , Cooke CL, Hansen LM, Lam AM, Gaddy JA, Johnson EM, Cariaga TA, Suarez G, Peek RM Jr, Cover TL, Solnick JV: Functional plasticity in the type IV secretion system of Helicobacter pylori. PLoS Pathog 9(2), e1003189 (2013)

    • Search Google Scholar
    • Export Citation
  • 16.

    Kumar Pachathundikandi S , Brandt S, Madassery J, Backert S: Induction of TLR-2 and TLR-5 expression by Helicobacter pylori switches cagPAI-dependent signalling leading to the secretion of IL-8 and TNF-alpha. PLoS One 6(5), e19614 (2011)

    • Search Google Scholar
    • Export Citation
  • 17.

    Nozawa Y , Nishihara K, Peek RM, Nakano M, Uji T, Ajioka H, Matsuura N, Miyake H: Identification of a signaling cascade for interleukin-8 production by Helicobacter pylori in human gastric epithelial cells. Biochem Pharmacol 64(1), 2130 (2002)

    • Search Google Scholar
    • Export Citation
  • 18.

    Owen RJ , Sharp S, Lawson AJ, Durrani Z, Rijpkema S, Kidd M: Investigation of the biological relevance of Helicobacter pylori cagE locus diversity, presence of CagA tyrosine phosphorylation motifs and vacuolating cytotoxin genotype on IL-8 induction in gastric epithelial cells. FEMS Immunol Med Microbiol 36(3), 135140 (2003)

    • Search Google Scholar
    • Export Citation
  • 19.

    Rieder G , Hatz RA, Moran AP, Walz A, Stolte M, Enders G: Role of adherence in interleukin-8 induction in Helicobacter pylori-associated gastritis. Infect Immun 65(9), 36223630 (1997)

    • Search Google Scholar
    • Export Citation
  • 20.

    Viala J , Chaput C, Boneca IG, Cardona A, Girardin SE, Moran AP, Athman R, Mémet S, Huerre MR, Coyle AJ, DiStefano PS, Sansonetti PJ, Labigne A, Bertin J, Philpott DJ, Ferrero RL: Nod1 responds to peptidoglycan delivered by the Helicobacter pylori cag pathogenicity island. Nat Immunol 5(11), 11661174 (2004)

    • Search Google Scholar
    • Export Citation
  • 21.

    Irving AT , Mimuro H, Kufer TA, Lo C, Wheeler R, Turner LJ, Thomas BJ, Malosse C, Gantier MP, Casillas LN, Votta BJ, Bertin J, Boneca IG, Sasakawa C, Philpott DJ, Ferrero RL, Kaparakis-Liaskos M: The immune receptor NOD1 and kinase RIP2 interact with bacterial peptidoglycan on early endosomes to promote autophagy and inflammatory signaling. Cell Host Microbe 15(5), 623635 (2014)

    • Search Google Scholar
    • Export Citation
  • 22.

    Aihara M , Tsuchimoto D, Takizawa H, Azuma A, Wakebe H, Ohmoto Y, Imagawa K, Kikuchi M, Mukaida N, Matsushima K: Mechanisms involved in Helicobacter pylori-induced interleukin-8 production by a gastric cancer cell line, MKN45. Infect Immun 65(8), 32183224 (1997)

    • Search Google Scholar
    • Export Citation
  • 23.

    Brandt S , Kwok T, Hartig R, Konig W, Backert S: NFkappaB activation and potentiation of proinflammatory responses by the Helicobacter pylori CagA protein. Proc Natl Acad Sci, U S A 102(26), 93009305 (2005)

    • Search Google Scholar
    • Export Citation
  • 24.

    Chu SH , Kim H, Seo JY, Lim JW, Mukaida N, Kim KH: Role of NF-kappaB and AP-1 on Helicobater pyloriinduced IL-8 expression in AGS cells. Dig Dis Sci 48(2), 257265 (2003)

    • Search Google Scholar
    • Export Citation
  • 25.

    Seo JH , Lim JW, Kim H, Kim KH: Helicobacter pylori in a Korean isolate activates mitogen-activated protein kinases, AP-1, and NF-kappaB and induces chemokine expression in gastric epithelial AGS cells. Lab Invest 84(1), 4962 (2004)

    • Search Google Scholar
    • Export Citation
  • 26.

    Naumann M , Wessler S, Bartsch C, Wieland B, Covacci A, Haas R, Meyer TF: Activation of activator protein 1 and stress response kinases in epithelial cells colonized by Helicobacter pylori encoding the cag pathogenicity island. J Biol Chem 274(44), 3165531662 (1999)

    • Search Google Scholar
    • Export Citation
  • 27.

    Sharma SA , Tummuru MK, Blaser MJ, Kerr LD: 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(5), 24012407 (1998)

    • Search Google Scholar
    • Export Citation
  • 28.

    Kwok T , Zabler D, Urman S, Rohde M, Hartig R, Wessler S, Misselwitz R, Berger J, Sewald N, König W, Backert S: Helicobacter exploits integrin for type IV secretion and kinase activation. Nature 449(7164), 862866 (2007)

    • Search Google Scholar
    • Export Citation
  • 29.

    Pham KT , Weiss E, Jimenez Soto LF, Breithaupt U, Haas R, Fischer W: CagI is an essential component of the Helicobacter pylori Cag type IV secretion system and forms a complex with CagL. PLoS One 7(4), e35341 (2012)

    • Search Google Scholar
    • Export Citation
  • 30.

    Gebert B , Fischer W, Weiss E, Hoffmann R, Haas R: Helicobacter pylori vacuolating cytotoxin inhibits T lymphocyte activation. Science 301(5636), 10991102 (2003)

    • Search Google Scholar
    • Export Citation
  • 31.

    Yasumoto K , Okamoto S, Mukaida N, Murakami S, Mai M, Matsushima K: Tumor necrosis factor alpha and interferon gamma synergistically induce interleukin 8 production in a human gastric cancer cell line through acting concurrently on AP-1 and NF-kB-like binding sites of the interleukin 8 gene. J Biol Chem 267(31), 2250622511 (1992)

    • Search Google Scholar
    • Export Citation
  • 32.

    Laemmli UK : Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259), 680685 (1970)

  • 33.

    Odenbreit S , Puls J, Sedlmaier B, Gerland E, Fischer W, Haas R: Translocation of Helicobacter pylori CagA into gastric epithelial cells by type IV secretion. Science 287(5457), 14971500 (2000)

    • Search Google Scholar
    • Export Citation
  • 34.

    Selbach M , Moese S, Meyer TF, Backert S: Functional analysis of the Helicobacter pylori cag pathogenicity island reveals both VirD4-CagA-dependent and VirD4-CagA-independent mechanisms. Infect Immun 70(2), 665671 (2002)

    • Search Google Scholar
    • Export Citation
  • 35.

    Naumann M : Pathogenicity island-dependent effects of Helicobacter pylori on intracellular signal transduction in epithelial cells. Int, J Med Microbiol 295(5), 335341 (1995)

    • Search Google Scholar
    • Export Citation
  • 36.

    Huang J , O’Toole PW, Doig P, Trust TJ: Stimulation of interleukin-8 production in epithelial cell lines by Helicobacter pylori. Infect Immun 63(5), 17321738 (1995)

    • Search Google Scholar
    • Export Citation
  • 37.

    Linden S , Nordman H, Hedenbro J, Hurtig M, Boren T, Carlstedt I: Strain-and blood group-dependent binding of Helicobacter pylori to human gastric MUC5AC glycoforms. Gastroenterology 123(6), 19231930 (2002)

    • Search Google Scholar
    • Export Citation
  • 38.

    Mahdavi J , Sonden B, Hurtig M, Olfat FO, Forsberg L, Roche N, Angstrom J, Larsson T, Teneberg S, Karlsson KA, Altraja S, Wadström T, Kersulyte D, Berg DE, Dubois A, Petersson C, Magnusson KE, Norberg T, Lindh F, Lundskog BB, Arnqvist A, Hammarström L, Borén T. Helicobacter pylori SabA adhesin in persistent infection and chronic inflammation. Science 297(5581), 573578 (2002)

    • Search Google Scholar
    • Export Citation
  • 39.

    Odenbreit S , Till M, Hofreuter D, Faller G, Haas R: Genetic and functional characterization of the alpAB gene locus essential for the adhesion of Helicobacter pylori to human gastric tissue. Mol Microbiol 31(5), 15371548 (1999)

    • Search Google Scholar
    • Export Citation
  • 40.

    Conradi J , Tegtmeyer N, Wozna M, Wissbrock M, Michalek C, Gagell C, Cover TL, Frank R, Sewald N, Backert S: An RGD helper sequence in CagL of Helicobacter pylori assists in interactions with integrins and injection of CagA. Front Cell Infect Microbiol 2, 70 (2012)

    • Search Google Scholar
    • Export Citation
  • 41.

    Jones DT : Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol 292(2), 195202 (1999)

  • 42.

    Mitra SK , Schlaepfer DD: Integrin-regulated FAK-Src signaling in normal and cancer cells. Curr Opin Cell Biol 18(5), 516523 (2006)

  • 43.

    Nilsson C , Sillen A, Eriksson L, Strand ML, Enroth H, Normark S, Falk P, Engstrand L: Correlation between cag pathogenicity island composition and Helicobacter pylori-associated gastroduodenal disease. Infect Immun 71(11), 65736578 (2003)

    • Search Google Scholar
    • Export Citation
  • 44.

    Covacci A , Rappuoli R: Tyrosine-phosphorylated bacterial proteins: Trojan horses for the host cell. J Exp Med 191(4), 587592 (2000)

  • 45.

    Backert S , Fronzes R, Waksman G: VirB2 and VirB5 proteins: specialized adhesins in bacterial type-IV secretion systems? Trends Microbiol 16(9), 409413 (2008)

    • Search Google Scholar
    • Export Citation
  • 46.

    Barden S , Lange S, Tegtmeyer N, Conradi J, Sewald N, Backert S, Niemann HH: A helical RGD motif promoting cell adhesion: crystal structures of the Helicobacter pylori type IV secretion system pilus protein CagL: structure 21(11), 19311941 (2013)

    • Search Google Scholar
    • Export Citation
  • 47.

    Kutter S , Buhrdorf R, Haas J, Schneider-Brachert W, Haas R, Fischer W: Protein subassemblies of the Helicobacter pylori Cag type IV secretion system revealed by localization and interaction studies. J Bacteriol 190(6), 21612171 (2008)

    • Search Google Scholar
    • Export Citation
  • 48.

    Strauss HM , Keller S: Pharmacological interference with protein–protein interactions mediated by coiled-coil motifs. Handb Exp Pharmacol 186), 461482 (2008)

    • Search Google Scholar
    • Export Citation
  • 49.

    Lupas A : Coiled coils: new structures and new functions. Trends Biochem Sci 21(10), 375382 (1996)

  • 50.

    Gazi AD , Charova SN, Panopoulos NJ, Kokkinidis M: Coiled-coils in type III secretion systems: structural flexibility, disorder and biological implications. Cell Microbiol 11(5), 719729 (2009)

    • Search Google Scholar
    • Export Citation
  • 51.

    Delahay RM , Frankel G: Coiled-coil proteins associated with type III secretion systems: a versatile domain revisited. Mol Microbiol 45(4), 905916 (2002)

    • Search Google Scholar
    • Export Citation
  • 52.

    Jimenez-Soto LF , Kutter S, Sewald X, Ertl C, Weiss E, Kapp U, et al.: Helicobacter pylori type IV secretion apparatus exploits beta1 integrin in a novel RGD-independent manner. PLoS Pathog 5(12), e1000684 (2009)

    • Search Google Scholar
    • Export Citation
  • 53.

    Grassl GA , Kracht M, Wiedemann A, Hoffmann E, Aepfelbacher M, von Eichel-Streiber C, Bohn E, Autenrieth IB: Activation of NF-kappaB and IL-8 by Yersinia enterocolitica invasin protein is conferred by engagement of Rac1 and MAP kinase cascades. Cell Microbiol 5(12), 957971 (2003)

    • Search Google Scholar
    • Export Citation
  • 54.

    Keates S , Keates AC, Katchar K, Peek RM Jr, Kelly CP: Helicobacter pylori induces up-regulation of the epidermal growth factor receptor in AGS gastric epithelial cells. J Infect Dis 196(1), 95103 (2007)

    • Search Google Scholar
    • Export Citation
  • 55.

    Joh T , Kataoka H, Tanida S, Watanabe K, Ohshima T, Sasaki M, et al.: Helicobacter pylori-stimulated interleukin-8 (IL-8) promotes cell proliferation through transactivation of epidermal growth factor receptor (EGFR) by disintegrin and metalloproteinase (ADAM) activation. Dig Dis Sci 50(11), 20812089 (2005)

    • Search Google Scholar
    • Export Citation
  • 56.

    Wallasch C , Crabtree JE, Bevec D, Robinson PA, Wagner H, Ullrich A: Helicobacter pylori-stimulated EGF receptor transactivation requires metalloprotease cleavage of HBEGF. Biochem Biophys Res Commun 295(3), 695701 (2002)

    • Search Google Scholar
    • Export Citation
  • 57.

    Backert S , Selbach M: Role of type IV secretion in Helicobacter pylori pathogenesis. Cell Microbiol 10(8), 15731581 (2008)

  • 58.

    Mimuro H , Berg DE, Sasakawa C: Control of epithelial cell structure and developmental fate: lessons from Helicobacter pylori. Bioessays 30(6), 515520 (2008)

    • Search Google Scholar
    • Export Citation

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