A Role for TLR4 in Infection and the Recognition of Surface Layer Proteins
Clostridium difficile is the etiological agent of antibiotic-associated diarrhoea (AAD) and pseudomembranous colitis in humans. The role of the surface layer proteins (SLPs) in this disease has not yet been fully explored. The aim of this study was to investigate a role for SLPs in the recognition of C. difficile and the subsequent activation of the immune system. Bone marrow derived dendritic cells (DCs) exposed to SLPs were assessed for production of inflammatory cytokines, expression of cell surface markers and their ability to generate T helper (Th) cell responses. DCs isolated from C3H/HeN and C3H/HeJ mice were used in order to examine whether SLPs are recognised by TLR4. The role of TLR4 in infection was examined in TLR4-deficient mice. SLPs induced maturation of DCs characterised by production of IL-12, TNFα and IL-10 and expression of MHC class II, CD40, CD80 and CD86. Furthermore, SLP-activated DCs generated Th cells producing IFNγ and IL-17. SLPs were unable to activate DCs isolated from TLR4-mutant C3H/HeJ mice and failed to induce a subsequent Th cell response. TLR4−/− and Myd88−/−, but not TRIF−/− mice were more susceptible than wild-type mice to C. difficile infection. Furthermore, SLPs activated NFκB, but not IRF3, downstream of TLR4. Our results indicate that SLPs isolated from C. difficile can activate innate and adaptive immunity and that these effects are mediated by TLR4, with TLR4 having a functional role in experimental C. difficile infection. This suggests an important role for SLPs in the recognition of C. difficile by the immune system.
Vyšlo v časopise:
A Role for TLR4 in Infection and the Recognition of Surface Layer Proteins. PLoS Pathog 7(6): e32767. doi:10.1371/journal.ppat.1002076
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002076
Souhrn
Clostridium difficile is the etiological agent of antibiotic-associated diarrhoea (AAD) and pseudomembranous colitis in humans. The role of the surface layer proteins (SLPs) in this disease has not yet been fully explored. The aim of this study was to investigate a role for SLPs in the recognition of C. difficile and the subsequent activation of the immune system. Bone marrow derived dendritic cells (DCs) exposed to SLPs were assessed for production of inflammatory cytokines, expression of cell surface markers and their ability to generate T helper (Th) cell responses. DCs isolated from C3H/HeN and C3H/HeJ mice were used in order to examine whether SLPs are recognised by TLR4. The role of TLR4 in infection was examined in TLR4-deficient mice. SLPs induced maturation of DCs characterised by production of IL-12, TNFα and IL-10 and expression of MHC class II, CD40, CD80 and CD86. Furthermore, SLP-activated DCs generated Th cells producing IFNγ and IL-17. SLPs were unable to activate DCs isolated from TLR4-mutant C3H/HeJ mice and failed to induce a subsequent Th cell response. TLR4−/− and Myd88−/−, but not TRIF−/− mice were more susceptible than wild-type mice to C. difficile infection. Furthermore, SLPs activated NFκB, but not IRF3, downstream of TLR4. Our results indicate that SLPs isolated from C. difficile can activate innate and adaptive immunity and that these effects are mediated by TLR4, with TLR4 having a functional role in experimental C. difficile infection. This suggests an important role for SLPs in the recognition of C. difficile by the immune system.
Zdroje
1. PoxtonIRMcCoubreyJBlairG 2001 The pathogenicity of Clostridium difficile. Clin Microbiol Infect 7 421 427
2. WrenBW 2006 Clostridium difficile comes of age. Future Microbiol 1 243 245
3. LyrasDO'ConnorJRHowarthPMSambolSPCarterGP 2009 Toxin B is essential for virulence of Clostridium difficile. Nature 458 1176 1179
4. BabcockGJBroeringTJHernandezHJMandellRBDonahueK 2006 Human monoclonal antibodies directed against toxins A and B prevent Clostridium difficile-induced mortality in hamsters. Infect Immun 74 6339 6347
5. Ni EidhinDBO'BrienJBMcCabeMSAthie-MoralesVKelleherDP 2008 Active immunization of hamsters against Clostridium difficile infection using surface-layer protein. FEMS Immunol Med Microbiol 52 207 218
6. O'BrienJBMcCabeMSAthie-MoralesVMcDonaldGSNi EidhinDB 2005 Passive immunisation of hamsters against Clostridium difficile infection using antibodies to surface layer proteins. FEMS Microbiol Lett 246 199 205
7. DrudyDCalabiEKyneLSougioultzisSKellyE 2004 Human antibody response to surface layer proteins in Clostridium difficile infection. FEMS Immunol Med Microbiol 41 237 242
8. Grogono-ThomasRDworkinJBlaserMJNewellDG 2000 Roles of the surface layer proteins of Campylobacter fetus subsp. fetus in ovine abortion. Infect Immun 68 1687 1691
9. SaraMSleytrUB 2000 S-Layer proteins. J Bacteriol 182 859 868
10. CarpenterSO'NeillLA 2007 How important are Toll-like receptors for antimicrobial responses? Cell Microbiol 9 1891 1901
11. AbelBThieblemontNQuesniauxVJBrownNMpagiJ 2002 Toll-like receptor 4 expression is required to control chronic Mycobacterium tuberculosis infection in mice. J Immunol 169 3155 3162
12. AkiraSTakedaK 2004 Toll-like receptor signalling. Nat Rev Immunol 4 499 511
13. BrikosCO'NeillLA 2008 Signalling of toll-like receptors. Handb Exp Pharmacol 21 50
14. JiangZGeorgelPDuXShamelLSovathS 2005 CD14 is required for MyD88-independent LPS signaling. Nat Immunol 6 565 570
15. KaganJCSuTHorngTChowAAkiraS 2008 TRAM couples endocytosis of Toll-like receptor 4 to the induction of interferon-beta. Nat Immunol 9 361 368
16. MoynaghPN 2005 TLR signalling and activation of IRFs: revisiting old friends from the NF-kappaB pathway. Trends Immunol 26 469 476
17. LoscherCEDraperELeavyOKelleherDMillsKH 2005 Conjugated linoleic acid suppresses NF-kappa B activation and IL-12 production in dendritic cells through ERK-mediated IL-10 induction. J Immunol 175 4990 4998
18. WeaverCTHattonRDManganPRHarringtonLE 2007 IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol 25 821 852
19. AndohMZhangGRussell-LodrigueKEShiveHRWeeksBR 2007 T cells are essential for bacterial clearance, and gamma interferon, tumor necrosis factor alpha, and B cells are crucial for disease development in Coxiella burnetii infection in mice. Infect Immun 75 3245 3255
20. WertsCTappingRIMathisonJCChuangTHKravchenkoV 2001 Leptospiral lipopolysaccharide activates cells through a TLR2-dependent mechanism. Nat Immunol 2 346 352
21. HoshinoKTakeuchiOKawaiTSanjoHOgawaT 1999 Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J Immunol 162 3749 3752
22. AdachiOKawaiTTakedaKMatsumotoMTsutsuiH 1998 Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. Immunity 9 143 150
23. YamamotoMSatoSHemmiHHoshinoKKaishoT 2003 Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301 640 643
24. SambolSPTangJKMerriganMMJohnsonSGerdingDN 2001 Infection of hamsters with epidemiologically important strains of Clostridium difficile. J Infect Dis 183 1760 1766
25. ChenXKatcharKGoldsmithJDNanthakumarNCheknisA 2008 A mouse model of Clostridium difficile-associated disease. Gastroenterology 135 1984 1992
26. KaurSVaishnaviCRayPKochharRPrasadKK
27. HigginsSCLavelleECMcCannCKeoghBMcNeelaE 2003 Toll-like receptor 4-mediated innate IL-10 activates antigen-specific regulatory T cells and confers resistance to Bordetella pertussis by inhibiting inflammatory pathology. J Immunol 171 3119 3127
28. LavelleECMcNeelaEArmstrongMELeavyOHigginsSC 2003 Cholera toxin promotes the induction of regulatory T cells specific for bystander antigens by modulating dendritic cell activation. J Immunol 171 2384 2392
29. LeeHKIwasakiA 2007 Innate control of adaptive immunity: dendritic cells and beyond. Semin Immunol 19 48 55
30. McLoughlinRMLeeJCKasperDLTzianabosAO 2008 IFN-gamma regulated chemokine production determines the outcome of Staphylococcus aureus infection. J Immunol 181 1323 1332
31. MilnerJDBrenchleyJMLaurenceAFreemanAFHillBJ 2008 Impaired T(H)17 cell differentiation in subjects with autosomal dominant hyper-IgE syndrome. Nature 452 773 776
32. ShutoTKatoKMoriYViriyakosolSObaM 2005 Membrane-anchored CD14 is required for LPS-induced TLR4 endocytosis in TLR4/MD-2/CD14 overexpressing CHO cells. Biochem Biophys Res Commun 338 1402 1409
33. FukataMMichelsenKSEriRThomasLSHuB 2005 Toll-like receptor-4 is required for intestinal response to epithelial injury and limiting bacterial translocation in a murine model of acute colitis. Am J Physiol Gastrointest Liver Physiol 288 G1055 1065
34. Rakoff-NahoumSPaglinoJEslami-VarzanehFEdbergSMedzhitovR 2004 Recognition of commensal microflora by toll-like receptors is required for intestinal homeostasis. Cell 118 229 241
35. AusielloCMCerquettiMFedeleGSpensieriFPalazzoR 2006 Surface layer proteins from Clostridium difficile induce inflammatory and regulatory cytokines in human monocytes and dendritic cells. Microbes Infect 8 2640 2646
36. FaganRPAlbesa-JoveDQaziOSvergunDIBrownKA 2009 Structural insights into the molecular organization of the S-layer from Clostridium difficile. Mol Microbiol 71 1308 1322
37. SiegemundSSchutzeNFreudenbergMALutzMBStraubingerRK 2007 Production of IL-12, IL-23 and IL-27p28 by bone marrow-derived conventional dendritic cells rather than macrophages after LPS/TLR4-dependent induction by Salmonella Enteritidis. Immunobiology 212 739 750
38. MahonBPSheahanBJGriffinFMurphyGMillsKH 1997 Atypical disease after Bordetella pertussis respiratory infection of mice with targeted disruptions of interferon-gamma receptor or immunoglobulin mu chain genes. J Exp Med 186 1843 1851
39. RudnerXLHappelKIYoungEAShellitoJE 2007 Interleukin-23 (IL-23)-IL-17 cytokine axis in murine Pneumocystis carinii infection. Infect Immun 75 3055 3061
40. HappelKIZhengMYoungEQuintonLJLockhartE 2003 Cutting edge: roles of Toll-like receptor 4 and IL-23 in IL-17 expression in response to Klebsiella pneumoniae infection. J Immunol 170 4432 4436
41. LawleyTDClareSWalkerAWGouldingDStablerRA 2009 Antibiotic treatment of clostridium difficile carrier mice triggers a supershedder state, spore-mediated transmission, and severe disease in immunocompromised hosts. Infect Immun 77 3661 3669
42. JordanJMWoodsMEOlanoJWalkerDH 2008 The absence of Toll-like receptor 4 signaling in C3H/HeJ mice predisposes them to overwhelming rickettsial infection and decreased protective Th1 responses. Infect Immun 76 3717 3724
43. O'NeillLA 2006 How Toll-like receptors signal: what we know and what we don't know. Curr Opin Immunol 18 3 9
44. McCoyCEO'NeillLA 2008 The role of toll-like receptors in macrophages. Front Biosci 13 62 70
45. Mata-HaroVCekicCMartinMChiltonPMCasellaCR 2007 The vaccine adjuvant monophosphoryl lipid A as a TRIF-biased agonist of TLR4. Science 316 1628 1632
46. BrunPScarpaMGrilloAPaluGMengoliC 2008 Clostridium difficile TxAC314 and SLP-36 kDa enhance the immune response toward a co-administered antigen. J Med Microbiol 57 725 731
47. PechineSJanoirCBoureauHGleizesATsapisN 2007 Diminished intestinal colonization by Clostridium difficile and immune response in mice after mucosal immunization with surface proteins of Clostridium difficile. Vaccine 25 3946 3954
48. HarrisGKuoLeeRChenW 2006 Role of Toll-like receptors in health and diseases of gastrointestinal tract. World J Gastroenterol 12 2149 2160
49. EidhinDNRyanAWDoyleRMWalshJBKelleherD 2006 Sequence and phylogenetic analysis of the gene for surface layer protein, slpA, from 14 PCR ribotypes of Clostridium difficile. J Med Microbiol 55 69 83
50. CerquettiMMolinariASebastianelliADiociaiutiMPetruzzelliR 2000 Characterization of surface layer proteins from different Clostridium difficile clinical isolates. Microb Pathog 28 363 372
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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