A lipid A-associated protein of Porphyromonas gingivalis, derived from the haemagglutinating domain of the RI protease gene family, is a potent stimulator of interleukin 6 synthesis Free

Abstract

SUMMARY: There is evidence that the lipid A-associated proteins (LAPs) of enteric bacteria can induce the synthesis of interleukin 1 (IL-1) and therefore may be important virulence factors. Porphyromonas gingivdis is now recognized as a major pathogen in the chronic inflammatory periodontal diseases and it has previously been reported that a crude LAP fraction from this organism could induce IL-1 and interleukin 6 (IL-6) synthesis. In the present study the chemical and biological properties of the LAPs of this bacterium have been further characterized. Analysis by SDS-PAGE has shown that the LAPs comprise nine proteins of molecular masses 81,68,48,47,28,25,20,17 and 16 kDa. These LAPS, at concentrations as low as 100 ng mV, were shown to stimulate human gingival fibroblasts, human peripheral blood mononuclear cells and whole human blood t o produce the pro-inflammatory cytokine IL-6. The cytokine-inducing activity of the LAPs was reduced after heat-inactivation and trypsinization, suggesting that the activity was not due to contaminating LPS. To establish which proteins in this mixture were the active cytokine inducers, the LAPs were separated by electrophoresis on polyacrylamide gels. The majority of the activity was associated with the 17 kDa LAP. N-terminal sequence analysis demonstrated that this protein was homologous t o an internal region of a conserved adhesin domain contained within a family of P. gingivdis extracellular proteins including the RI protease, Lys-gingipain, porphypain and haemagglutinin A. In addition to a role in adherence, the adhesin domain(s) of these proteins may also have cytokine-inducing properties. Furthermore, it has also been shown that the previously observed degradation of cytokines by P. gingivelis may be attributable to the catalytic domain of the RI protease. Thus, different domains within the same molecule appear t o have opposing actions on pro-inflammatory cytokine levels and the balance between these two activities may influencethe cytokine status of the periodontiurn in patients with the common chronic inflammatory conditions known as the periodontal diseases.

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1998-11-01
2024-03-28
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References

  1. Aduse-Opoku J., Muir J., Slaney J.M., Rangarajan M., Curtis M.A. 1995; Characterization, genetic analysis, and expression of a protease antigen (PrpRI) of Porphyromonas gingivalis W50.. Infect Immun 63:4744–4754
    [Google Scholar]
  2. Aduse-Opoku J., Slaney J.M., Rangarajan M., Muir J., Young K.A., Curtis M.A. 1997; The Tla of Porphyromonas gingivalis W50: a homologue of the arginine-specific protease precursor (PrpRI) which shares sequence similarity to TonB linked receptors.. J Bacteriol 179:4778–4788
    [Google Scholar]
  3. Barkocy-Gallagher G.A., Han N., Patti J.M., Whitlock J., Progulske-Fox A., Lantz M.S. 1996; Analysis of the prtP gene encoding porphypain, a cysteine proteinase of Porphyromonas gingivalis.. J Bacteriol 178:2734–2741
    [Google Scholar]
  4. Bjornson B.H., Agura E., Harvey J., Johns M., Andrews R.G., McCabe W.R. 1988; Endotoxin-associated protein; a potent stimulus for human granulopoietic activity which may be accessory cell independent.. Infect Immun 56:1602–1607
    [Google Scholar]
  5. Calkins C.C., Platt K., Potempa J., Travis J. 1998; Inactivation of tumor necrosis factor-alpha by proteinases (gingipains) from the periodontal pathogen, Porphyromonas gingivalis. Implications of immune system evasion.. J Biol Chem 273:6611–6614
    [Google Scholar]
  6. Cridland J.C., Booth V., Ashley F.P., Curtis M.A., Wilson R.F., Shepherd P. 1994; Preliminary characterisation of antigens recognised by monoclonal antibodies raised to Porphyromonas gingivalis and by sera from patients with periodontitis.. J Periodontal Res 29:339–347
    [Google Scholar]
  7. Cunningham M.D., Seachford C., Ratcliffe K., Bainbridge B., Aruffo A., Darveau R.P. 1996; Helicobacter pylori and Porphyromonas gingivalis lipopolysaccharides are poorly transferred to recombinant soluble CD14.. Infect Immun 64:3601–3608
    [Google Scholar]
  8. Curtis M.A., Aduse-Opoku J., Slaney J.M., Rangarajan M., Booth V., Cridland J., Shepherd P. 1996; Characterization of an adherence and antigenic determinant of the ArgI protease of Porphyromonas gingivalis which is present on multiple gene products.. Infect Immun 64:2532–2539
    [Google Scholar]
  9. Darveau R.P., Hancock R.E.W. 1983; Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains.. J Bacteriol 155:831–838
    [Google Scholar]
  10. Emancipator K., Csako G., Elin R.J. 1996; In vitro inactivation of bacterial endotoxin by human lipoproteins and apoproteins.. Infect Immun 60:596–601
    [Google Scholar]
  11. European Pharmacopoeia 1997, Third edition. 2.6.14.. Bacterial Endotoxins pp. 29–37 Strasbourg: European Department for the Quality of Medicines;
    [Google Scholar]
  12. Fletcher J., Reddi K., Poole S., Nair S., Henderson B., Tabona P., Wilson M. 1997; Interactions between periodonto- pathogenic bacteria and cytokines.. J Periodontal Res 32:200–205
    [Google Scholar]
  13. Fletcher J., Nair S., Poole S., Henderson B., Wilson M. 1998; Cytokine degradation by biofilms of Porphyromonas gingivalis.. Curr Microbiol 36:216–219
    [Google Scholar]
  14. Goodman G.W., Sultzer B.M. 1979a; Characterization of the chemical and physical properties of a novel B-lymphocyte activator, endotoxin protein.. Infect Immun 24:685–696
    [Google Scholar]
  15. Goodman G.W., Sultzer B.M. 1979b; Further studies on the activation of lymphocytes by endotoxin protein.. J Immunol 122:1329–1334
    [Google Scholar]
  16. Hamada S., Toga T., Nishihara T., Fujiwara T., Okahashi N. 1988; Characterization and immunobiological activities of lipopolysaccharides from periodontal bacteria.. Adv Dent Res 2:284–291
    [Google Scholar]
  17. Hitchcock P.J., Morrison D.C. 1984; The protein component of bacterial endotoxins.. In Handbook of Endotoxin 1 Chemistry of Endotoxin pp. 339–374 Rietschel E.T., Bhattacharya SK. (editors) Amsterdam: Elsevier;
    [Google Scholar]
  18. Hogan M.M., Vogel S.N. 1987; Lipid A-associated proteins provide an alternative “second signal” in the activation of recombinant interferon-gamma-primed C3H/HeJ macrophages to a fully tumoricidal state.. J Immunol 139:3697–3702
    [Google Scholar]
  19. Hogan M.M., Vogel S.N. 1988; Production of tumor necrosis factor by rIFN-gamma-primed C3H/HeJ (Lpsd) macrophages requires the presence of lipid A-associated proteins.. J Immunol 141:4196–4202
    [Google Scholar]
  20. Johns M.A., Sipe J.D., Melton L.B., Strom T.B., McCabe W.R. 1988; Endotoxin-associated protein; interleukin-l-like activity on serum amyloid A synthesis and T-lymphocyte activation.. Infect Immun 56:1593–1601
    [Google Scholar]
  21. Killion J.W., Morrison D.C. 1986; Protection of C3H/HeJ mice from lethal Salmonella typhimurium LT2 infection by immunization with lipopolysaccharide-lipid A-associated protein complexes.. Infect Immun 54:1–8
    [Google Scholar]
  22. Kurimatsu H.K., Yoneda M., Madden T. 1995; Proteases and collagenases of Porphyromonas gingivalis.. Adv Dent Res 9:37–40
    [Google Scholar]
  23. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  24. Lantz M.S., Allen R.D., Duck L.W., Switalski L.M., Hook M. 1991; Porphyromonas gingivalis surface components bind and degrade connective tissue proteins.. J Periodontal Res 26:283–285
    [Google Scholar]
  25. Mangan D.F., Wahl S.M., Sultzer B.M., Mergenhagen S.E. 1993; Stimulation of human monocytes by endotoxin-associated protein; inhibition of programmed cell death (apoptosis) and potential significance in adjuvancy.. Infect lmmun 60:1648–1686
    [Google Scholar]
  26. Matsudaira P. 1987; Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.. J Biol Chem 262:10035–10038
    [Google Scholar]
  27. Morrison D.C., Jacobs D.M. 1976; Binding of polymyxin B to the lipid-A region of bacterial LPS.. Immunochemistry 13:815–818
    [Google Scholar]
  28. Morrison D.C., Leive L. 1975; Fractions of lipopoly- saccharides from E. coli 0111 ;B4 prepared by two extraction procedures.. J Biol Chem 250:2911–2919
    [Google Scholar]
  29. Naiming H.A., Whitlock J., Progulske-Fox A. 1996; Thehemagglutinin gene A (hagA) of Porphyromonas gingivalis 381 contains four large contiguous direct repeats.. Infect lmmun 64:4000–4007
    [Google Scholar]
  30. Pavloff N., Potempa J., Pike R.N., Prochazka V., Kiefer M.C., Travis J., Barr P.J. 1995; Molecular cloning and structural characterization of the Arg-gingipain proteinase of Porphyro-monas gingivalis. Biosynthesis as a proteinase-adhesin poly-protein.. J Biol Chem 270:1007–1010
    [Google Scholar]
  31. Phillips M., Castagna R., Sultzer B.M., Eisenstein T.K. 1989; Immunogenic endotoxin associated protein from a rough strain of Salmonella.. FEMS Microbiol Immunol 1:485–490
    [Google Scholar]
  32. Pike R., McGraw W., Potempa J., Travis J. 1994; Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization and evidence for the existence of complexes with hemagglutinins.. J Biol Chem 269:406–411
    [Google Scholar]
  33. Porat R., Yanoov M., Johns M.A., Shibolet S., Michalevicz R. 1992; Effects of endotoxin-associated protein on hematopoiesis.. Infect lmmun 60:1756–1760
    [Google Scholar]
  34. Progulske-Fox A., Rao V., Han N., Lepine G., Witlock J., Lantz M. 1993; Molecular characterization of hemagglutinin genes of periodontopathic bacteria.. J Periodontal Res 28:473–474
    [Google Scholar]
  35. Reddi K., Meghji S., Wilson M., Henderson B. 1995a; Comparison of the osteolytic activity of surface-associated proteins of bacteria implicated in periodontal disease.. Oral Dis 1:26–31
    [Google Scholar]
  36. Reddi K., Poole S., Nair S., Meghji S., Henderson B., Wilson M. 1995b; Lipid A-associated proteins from periodonto- pathogenic bacteria induce interleukin-6 production by human gingival fibroblasts and monocytes.. FEMS Immunol Med Microbiol 11:137–144
    [Google Scholar]
  37. Reife R.A., Shapiro R.A., Bamber B.A., Berry K.K., Mick G.E., Darveau R.P. 1995; Porphyromonas gingivalis lipopoly- saccharide is poorly recognized by molecular components of nnate host defence in a mouse model of early inflammation.. Infect lmmun 63:4686–4694
    [Google Scholar]
  38. Sultzer B.M. 1968; Genetic control in leukocyte responses to endotoxin.. Nature 219:1253–1254
    [Google Scholar]
  39. Sultzer B.M. 1969; Genetic factors in leukocyte responses to endotoxin; further studies in mice.. J Immunol 103:32–38
    [Google Scholar]
  40. Sultzer B.M., Goodman G.W. 1976; Endotoxin protein; a B- cell mitogen and polyclonal activator of C3H/HeJ lymphocytes.. J Exp Med 144:821–827
    [Google Scholar]
  41. Sultzer B.M., Craig J.P., Castagna R. 1985; The adjuvant effect of pertussis endotoxin protein in modulating the immune response to cholera toxoid in mice.. Dev Biol Stand 61:225–232
    [Google Scholar]
  42. Taktak Y.S., Selkirk S., Bristow A.F., Carpenter A., Ball C., Rafferty B., Poole S. 1991; Assay of pyrogens by interleukin- 6 release from monocytic cell lines.. J Pharm Pharmacol 43:578–582
    [Google Scholar]
  43. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets; procedure and some applications.. Proc Natl Acad Sci USA 76:4350–4354
    [Google Scholar]
  44. Viriyakosol S., Kirkland T.N. 1996; The N-terminal half of membrane CD 14 is a functional cellular lipopolysaccharide receptor.. Infect lmmun 64:653–656
    [Google Scholar]
  45. Watanabe A., Takeshita A., Kitano S., Hanazawa S. 1996; CD 14 mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in HGFs.. Infect lmmun 11:4484–4494
    [Google Scholar]
  46. Westphal O., Jann K. 1965; Bacterial lipopolysaccharide. Extraction with phenol-water and further applications of the procedure.. Methods Carbohydr Chem 5:83–91
    [Google Scholar]
  47. Wilson M., Meghji S., Barber P., Henderson B. 1993; Biological activities of surface-associated material from Por-phyromonas gingivalis.. FEMS Immunol Med Microbiol 6:147–155
    [Google Scholar]
  48. Wober W., Alaupovic P. 1971; Studies on the protein moiety of endotoxin from gram-negative bacteria. Characterization of the protein moiety isolated by phenol treatment of endotoxin from Serratia marcescens 08 and Escherichia coli 0 141;K85(B).. Eur J Biochem 19:340–356
    [Google Scholar]
  49. Wurfel M.M., Kunitake S.T., Lichenstein H., Kane J.P., Wright S.D. 1994; Lipopolysaccharide-binding protein is carried on lipoproteins and acts as a co-factor in the neutralization of LPS.. J Exp Med 180:1025–1035
    [Google Scholar]
  50. Wurfel M.M., Hailman E., Wright S.D. 1995; Soluble CD 14 acts as a shuttle in the neutralization of LPS by LPS binding protein and reconstituted lipoprotein.. J Exp Med 181:1743–1745
    [Google Scholar]
  51. Ziegler-Heitbrock H.W., Frankenberger H., Wedel A. 1995; Tolerance to lipopolysaccharide in human blood monocytes.. Immunobiology 193:217–223
    [Google Scholar]
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