Interferon-gamma secretion is induced in IL-12 stimulated human NK cells by recognition of Helicobacter pylori or TLR2 ligands

Abstract

Helicobacter pylori induce a chronic inflammation in the human gastric mucosa characterized by increased production of interferon-gamma (IFN-γ). The presence of natural killer (NK) cells in the human gastric mucosa and the ability of NK cells to produce IFN-γ suggest an important role of NK cells in the immune response directed towards H. pylori infection. Since NK cells previously have been shown to respond to bacterial components with IFN-γ production, we investigated the mechanisms for the recognition of H. pylori. We found that inhibition of MyD88 homodimerization resulted in decreased production of IFN-γ and that inhibition of the p38 MAPK decreased the production as well as the secretion of IFN-γ. Further studies indicated an involvement of Toll-like receptors (TLRs), in particular TLR2. Finally, we showed that the H. pylori specific membrane bound lipoprotein HpaA induced IFN-γ production from NK cells through recognition by TLR2. In conclusion, we suggest an involvement of TLR2 in the recognition of H. pylori by human NK cells and that HpaA is a TLR2 ligand important for recognition.

Keywords

Human NK cells bacterial infection cell surface molecules cytokines H pylori IFN-γ

References

Byrne P. , McGuirk P. , Todryk S. , Mills KH Depletion of NK cells results in disseminating lethal infection with Bordetella pertussis associated with a reduction of antigen-specific Th1 and enhancement of Th2, but not Tr1 cells. Eur J Immunol 2004 ; 34: 2579-2588.

Sporri R. , Joller N. , Albers U. , Hilbi H. , Oxenius A. MyD88-dependent IFN-gamma production by NK cells is key for control of Legionella pneumophila infection. J Immunol 2006; 176: 6162-6171.

Papamichail M. , Perez SA , Gritzapis AD , Baxevanis CN Natural killer lymphocytes: biology, development, and function. Cancer Immunol Immunother 2004 ; 53: 176-186.

Cerwenka A. , Lanier LL Natural killer cells, viruses and cancer. Nat Rev Immunol 2001; 1: 41-49.

Biron CA , Nguyen KB , Pien GC , Cousens LP , Salazar-Mather TP Natural killer cells in antiviral defense: function and regulation by innate cytokines. Annu Rev Immunol 1999; 17: 189-220.

Sugiyama T. , Asaka M. Helicobacter pylori infection and gastric cancer. Med Electron Microsc 2004; 37: 149-157.

Pellicano A. , Sebkova L. , Monteleone G. et al. Interleukin-12 drives the Th1 signaling pathway in Helicobacter pylori-infected human gastric mucosa. Infect Immun 2007; 75: 1738-1744.

Sommer F. , Faller G. , Konturek P. et al. Antrum- and corpus mucosa-infiltrating CD4(+) lymphocytes in Helicobacter pylori gastritis display a Th1 phenotype. Infect Immun 1998; 66: 5543-5546.

Trinchieri G. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat Rev Immunol 2003; 3: 133-146.

10.

Akhiani AA , Pappo J. , Kabok Z. et al. Protection against Helicobacter pylori infection following immunization is IL-12-dependent and mediated by Th1 cells. J Immunol 2002; 169: 6977-6984.

11.

Yun CH , Lundgren A. , Azem J. et al. Natural killer cells and Helicobacter pylori infection: bacterial antigens and interleukin-12 act synergistically to induce gamma interferon production. Infect Immun 2005; 73: 1482-1490.

12.

Hayakawa Y. , Takeda K. , Yagita H. et al. IFN-gamma-mediated inhibition of tumor angiogenesis by natural killer T-cell ligand, alpha-galactosylceramide. Blood 2002; 100: 1728-1733.

13.

Ishigami S. , Natsugoe S. , Tokuda K. et al. Clinical impact of intratumoral natural killer cell and dendritic cell infiltration in gastric cancer. Cancer Lett 2000; 159: 103-108.

14.

Takeuchi H. , Maehara Y. , Tokunaga E. , Koga T. , Kakeji Y. , Sugimachi K. Prognostic significance of natural killer cell activity in patients with gastric carcinoma: a multivariate analysis . Am J Gastroenterol 2001; 96: 574-578.

15.

Helicobacter and Cancer Collaborative Group. Gastric cancer and Helicobacter pylori: a combined analysis of 12 case control studies nested within prospective cohorts. Gut 2001; 49: 347-353.

16.

HornungV, Rothenfusser S. , Britsch S. et al. Quantitative expression of Toll-like receptor 1-10 mRNA in cellular subsets of human peripheral blood mononuclear cells and sensitivity to CpG oligodeoxynucleotides. J Immunol 2002; 168: 4531-4537.

17.

Mandell L. , Moran AP , Cocchiarella A. et al. Intact Gram-negative Helicobacter pylori, Helicobacter felis, and Helicobacter hepaticus bacteria activate innate immunity via Toll-like receptor 2 but not Toll-like receptor 4. Infect Immun 2004; 72: 6446-6454.

18.

Torok AM , Bouton AH , Goldberg JB Helicobacter pylori induces interleukin-8 secretion by Toll-like receptor 2- and Toll-like receptor 5-dependent and -independent pathways. Infect Immun 2005; 73: 1523-1531.

19.

Ghiara P. , Rossi M. , Marchetti M. et al. Therapeutic intragastric vaccination against Helicobacter pylori in mice eradicates an otherwise chronic infection and confers protection against reinfection. Infect Immun 1997; 65: 4996-5002.

20.

Lundstrom AM , Bolin I. , Bystrom M. , Nystrom S. Recombinant HpaA purified from Escherichia coli has biological properties similar to those of native Helicobacter pylori HpaA. APMIS 2003; 111: 389-397.

21.

Lundin BS , Johansson C. , Svennerholm AM Oral immunization with a Salmonella enterica serovar typhi vaccine induces specific circulating mucosa-homing CD4(+) and CD8(+) T cells in humans. Infect Immun 2002; 70: 5622-5627.

22.

Strengell M. , Matikainen S. , Siren J. et al. IL-21 in synergy with IL-15 or IL-18 enhances IFN-gamma production in human NK and T cells. J Immunol 2003; 170: 5464-5469.

23.

Okusawa T. , Fujita M. , Nakamura J. et al. Relationship between structures and biological activities of mycoplasmal diacylated lipopeptides and their recognition by Toll-like receptors 2 and 6. Infect Immun 2004; 72: 1657-1665.

24.

Takeuchi O. , Sato S. , Horiuchi T. et al. Role of Toll-like receptor 1 in mediating immune response to microbial lipoproteins. J Immunol 2002; 169: 10-14.

25.

Andersen-Nissen E. , Smith KD , Strobe KL et al. Evasion of Toll-like receptor 5 by flagellated bacteria. Proc Natl Acad Sci USA 2005; 102: 9247-9252.

26.

Carlsohn E. , Nystrom J. , BolinI, Nilsson CL , Svennerholm AM HpaA is essential for Helicobacter pylori colonization in mice. Infect Immun 2006; 74: 920-926.

27.

Hirata Y. , Ohmae T. , Shibata W. et al. MyD88 and TNF receptor-associated factor 6 are critical signal transducers in Helicobacter pylori-infected human epithelial cells. J Immunol 2006; 176: 3796-3803.

28.

Pisegna S. , Pirozzi G. , Piccoli M. , Frati L. , Santoni A. , Palmieri G. p38 MAPK activation controls the TLR3-mediated up-regulation of cytotoxicity and cytokine production in human NK cells. Blood 2004; 104: 4157-4164.

29.

Girart MV , Fuertes MB , Domaica CI , Rossi LE , Zwirner NW Engagement of TLR3, TLR7, and NKG2D regulate IFN-gamma secretion but not NKG2D-mediated cytotoxicity by human NK cells stimulated with suboptimal doses of IL-12. J Immunol 2007; 179: 3472-3479.

30.

Sivori S. , Falco M. , Della Chiesa M. et al. CpG and double-stranded RNA trigger human NK cells by Toll-like receptors: induction of cytokine release and cytotoxicity against tumors and dendritic cells. Proc Natl Acad Sci USA 2004 ; 101: 10116-10121.

31.

Gorski KS , Waller EL , Bjornton-Severson J. et al. Distinct indirect pathways govern human NK-cell activation by TLR-7 and TLR-8 agonists. Int Immunol 2006; 18: 1115-1126.

32.

Schmidt KN , Leung B. , Kwong M. et al. APC-independent activation of NK cells by the Toll-like receptor 3 agonist double-stranded RNA. J Immunol 2004 ; 172: 138-143.

33.

Hart OM , Athie-Morales V. , O’Connor GM , Gardiner CM TLR7/ 8-mediated activation of human NK cells results in accessory cell-dependent IFN-gamma production. J Immunol 2005; 175: 1636-1642.

34.

Kamath AT , Sheasby CE , Tough DF Dendritic cells and NK cells stimulate bystander T cell activation in response to TLR agonists through secretion of IFN-alpha beta and IFN-gamma. J Immunol 2005; 174: 767-776.

35.

Smith Jr MF , Mitchell A. , Li G. et al. 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 2003; 278: 32552-32560.

36.

Marcenaro E. , Ferranti B. , Falco M. , Moretta L. , Moretta A. Human NK cells directly recognize Mycobacterium bovis via TLR2 and acquire the ability to kill monocyte-derived DC. Int Immunol 2008; 20: 1155-1167.

37.

BeckerI, Salaiza N. , Aguirre M. et al. Leishmania lipophosphoglycan (LPG) activates NK cells through Toll-like receptor-2. Mol Biochem Parasitol 2003; 130: 65-74.

38.

Eriksson M. , Meadows SK , Basu S. , Mselle TF , Wira CR , Sentman CL TLRs mediate IFN-gamma production by human uterine NK cells in endometrium. J Immunol 2006; 176: 6219-6224.

39.

Della Chiesa M. , Sivori S. , Castriconi R. , Marcenaro E. , Moretta A. Pathogen-induced private conversations between natural killer and dendritic cells. Trends Microbiol 2005; 13: 128-136.

40.

Hafsi N. , Voland P. , Schwendy S. et al. Human dendritic cells respond to Helicobacter pylori, promoting NK cell and Th1-effector responses in vitro. J Immunol 2004; 173: 1249-1257.

41.

Matysiak-Budnik T. , Coffin B. , Lavergne-Slove A. , Sabate JM , Megraud F. , Heyman M. Helicobacter pylori increases the epithelial permeability to a food antigen in human gastric biopsies . Am J Gastroenterol 2004; 99: 225-232.

42.

Ismail S. , Hampton MB , Keenan JI Helicobacter pylori outer membrane vesicles modulate proliferation and interleukin-8 production by gastric epithelial cells. Infect Immun 2003; 71: 5670-5675.

43.

Dubois A. , Boren T. Helicobacter pylori is invasive and it may be a facultative intracellular organism. Cell Microbiol 2007; 9: 1108-1116.

44.

Shegarfi H. , Sydnes K. , Lovik M. , Inngjerdingen M. , Rolstad B. , Naper C. The role of natural killer cells in resistance to the intracellular bacterium Listeria monocytogenes in rats. Scand J Immunol 2009; 70: 238-244.

45.

Le-Barillec K. , Magalhaes JG , Corcuff E. et al. Roles for T and NK cells in the innate immune response to Shigella flexneri. J Immunol 2005; 175: 1735-1740.

46.

Harrington L. , Srikanth CV , Antony R. , Shi HN , Cherayil BJ A role for natural killer cells in intestinal inflammation caused by infection with Salmonella enterica serovar Typhimurium . FEMS Immunol Med Microbiol 2007; 51: 372-380.

47.

Sayi A. , Kohler E. , Hitzler I. et al. The CD4+ T cell-mediated IFN-gamma response to Helicobacter infection is essential for clearance and determines gastric cancer risk. J Immunol 2009; 182: 7085-7101.