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. 1997 Aug;65(8):3248–3254. doi: 10.1128/iai.65.8.3248-3254.1997

Effects of Porphyromonas gingivalis and Escherichia coli lipopolysaccharides on mononuclear phagocytes.

F A Roberts 1, G J Richardson 1, S M Michalek 1
PMCID: PMC175459  PMID: 9234782

Abstract

The mononuclear phagocyte plays an important role in the regulation of microbe-induced inflammation, in part through its ability to secrete mediators, particularly cytokines, in response to microorganisms and their products. To evaluate the effects of the microbial flora associated with chronic adult periodontitis on cytokine induction, lipopolysaccharide (LPS) from the periodontopathogen Porphyromonas gingivalis was used to stimulate naive and phorbol ester-primed U937 monocytic cells, as well as elutriated human peripheral blood monocytes. We assessed the effect of this LPS, in comparison to that of LPS from Escherichia coli, on cell proliferation, cytokine induction, and surface expression of the LPS receptor CD14. P. gingivalis LPS stimulated proliferation of U937 cells at concentrations of greater than 1 ng/ml, while E. coli LPS inhibited proliferation. Phorbol myristic acid (PMA)-treated U937 cells and elutriated monocytes responded to E. coli LPS activation by producing tumor necrosis factor alpha (TNF-alpha) mRNA and protein; however, P. gingivalis LPS induced greater numbers of TNF-alpha mRNA-positive cells and higher (P < 0.05) levels of protein than did E. coli LPS. Both cell types expressed interleukin-1 beta (IL-1beta) mRNA and protein in response to either LPS treatment. Compared with E. coli LPS, P. gingivalis LPS induced significantly (P < 0.05) higher numbers of IL-1 mRNA-positive U937 cells and elutriated monocytes, as well as production of significantly more (P < 0.05) IL-1 protein by the monocytes. The PMA-treated U937 cells and the monocytes produced high levels of IL-1 receptor antagonist mRNA and protein which were only marginally affected by the LPS preparations. While E. coli LPS induced expression of CD 14 on the surface of PMA-primed U937 cells and monocytes, P. gingivalis LPS exhibited a significantly (P < 0.05) greater ability to enhance receptor levels. Our results indicate that P. gingivalis LPS can activate the mononuclear phagocyte for proliferation, cytokine production, and CD14 expression, providing evidence for the potential of this bacterial component to act as a critical regulatory factor in the chronic inflammatory response associated with periodontitis.

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Selected References

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  1. Arend W. P. Interleukin 1 receptor antagonist. A new member of the interleukin 1 family. J Clin Invest. 1991 Nov;88(5):1445–1451. doi: 10.1172/JCI115453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beutler B. TNF, immunity and inflammatory disease: lessons of the past decade. J Investig Med. 1995 Jun;43(3):227–235. [PubMed] [Google Scholar]
  3. Bone R. C. The pathogenesis of sepsis. Ann Intern Med. 1991 Sep 15;115(6):457–469. doi: 10.7326/0003-4819-115-6-457. [DOI] [PubMed] [Google Scholar]
  4. Bucy R. P., Karr L., Huang G. Q., Li J., Carter D., Honjo K., Lemons J. A., Murphy K. M., Weaver C. T. Single cell analysis of cytokine gene coexpression during CD4+ T-cell phenotype development. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7565–7569. doi: 10.1073/pnas.92.16.7565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burchett S. K., Weaver W. M., Westall J. A., Larsen A., Kronheim S., Wilson C. B. Regulation of tumor necrosis factor/cachectin and IL-1 secretion in human mononuclear phagocytes. J Immunol. 1988 May 15;140(10):3473–3481. [PubMed] [Google Scholar]
  6. Daniel-Issakani S., Spiegel A. M., Strulovici B. Lipopolysaccharide response is linked to the GTP binding protein, Gi2, in the promonocytic cell line U937. J Biol Chem. 1989 Dec 5;264(34):20240–20247. [PubMed] [Google Scholar]
  7. Darveau R. P., Cunningham M. D., Bailey T., Seachord C., Ratcliffe K., Bainbridge B., Dietsch M., Page R. C., Aruffo A. Ability of bacteria associated with chronic inflammatory disease to stimulate E-selectin expression and promote neutrophil adhesion. Infect Immun. 1995 Apr;63(4):1311–1317. doi: 10.1128/iai.63.4.1311-1317.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dinarello C. A. The interleukin-1 family: 10 years of discovery. FASEB J. 1994 Dec;8(15):1314–1325. [PubMed] [Google Scholar]
  9. Ertel W., Morrison M. H., Wang P., Ba Z. F., Ayala A., Chaudry I. H. The complex pattern of cytokines in sepsis. Association between prostaglandins, cachectin, and interleukins. Ann Surg. 1991 Aug;214(2):141–148. doi: 10.1097/00000658-199108000-00008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gemmell E., Seymour G. J. Different responses in B cells induced by Porphyromonas gingivalis and Fusobacterium nucleatum. Arch Oral Biol. 1992;37(7):565–573. doi: 10.1016/0003-9969(92)90139-y. [DOI] [PubMed] [Google Scholar]
  11. Genco R. J. Host responses in periodontal diseases: current concepts. J Periodontol. 1992 Apr;63(4 Suppl):338–355. doi: 10.1902/jop.1992.63.4s.338. [DOI] [PubMed] [Google Scholar]
  12. Hamada S., Takada H., Ogawa T., Fujiwara T., Mihara J. Lipopolysaccharides of oral anaerobes associated with chronic inflammation: chemical and immunomodulating properties. Int Rev Immunol. 1990;6(4):247–261. doi: 10.3109/08830189009056635. [DOI] [PubMed] [Google Scholar]
  13. Hammerberg C., Arend W. P., Fisher G. J., Chan L. S., Berger A. E., Haskill J. S., Voorhees J. J., Cooper K. D. Interleukin-1 receptor antagonist in normal and psoriatic epidermis. J Clin Invest. 1992 Aug;90(2):571–583. doi: 10.1172/JCI115896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hannum C. H., Wilcox C. J., Arend W. P., Joslin F. G., Dripps D. J., Heimdal P. L., Armes L. G., Sommer A., Eisenberg S. P., Thompson R. C. Interleukin-1 receptor antagonist activity of a human interleukin-1 inhibitor. Nature. 1990 Jan 25;343(6256):336–340. doi: 10.1038/343336a0. [DOI] [PubMed] [Google Scholar]
  15. Hass R., Bartels H., Topley N., Hadam M., Köhler L., Goppelt-Strübe M., Resch K. TPA-induced differentiation and adhesion of U937 cells: changes in ultrastructure, cytoskeletal organization and expression of cell surface antigens. Eur J Cell Biol. 1989 Apr;48(2):282–293. [PubMed] [Google Scholar]
  16. Howard M., Paul W. E. Regulation of B-cell growth and differentiation by soluble factors. Annu Rev Immunol. 1983;1:307–333. doi: 10.1146/annurev.iy.01.040183.001515. [DOI] [PubMed] [Google Scholar]
  17. Karr L. J., Panoskaltsis-Mortari A., Li J., Devore-Carter D., Weaver C. T., Bucy R. P. In situ hybridization for cytokine mRNA with digoxigenin-labeled riboprobes. Sensitivity of detection and double label applications. J Immunol Methods. 1995 May 11;182(1):93–106. doi: 10.1016/0022-1759(95)00027-8. [DOI] [PubMed] [Google Scholar]
  18. Kielian T. L., Blecha F. CD14 and other recognition molecules for lipopolysaccharide: a review. Immunopharmacology. 1995 Apr;29(3):187–205. doi: 10.1016/0162-3109(95)00003-c. [DOI] [PubMed] [Google Scholar]
  19. Klein J. B., Schepers T. M., Dean W. L., Sonnenfeld G., McLeish K. R. Role of intracellular calcium concentration and protein kinase C activation in IFN-gamma stimulation of U937 cells. J Immunol. 1990 Jun 1;144(11):4305–4311. [PubMed] [Google Scholar]
  20. Koga T., Nishihara T., Fujiwara T., Nisizawa T., Okahashi N., Noguchi T., Hamada S. Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli. Infect Immun. 1985 Mar;47(3):638–647. doi: 10.1128/iai.47.3.638-647.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mangan D. F., Mergenhagen S. E., Wahl S. M. Apoptosis in human monocytes: possible role in chronic inflammatory diseases. J Periodontol. 1993 May;64(5 Suppl):461–466. [PubMed] [Google Scholar]
  22. McIntire F. C., Sievert H. W., Barlow G. H., Finley R. A., Lee A. Y. Chemical, physical, biological properties of a lipopolysaccharide from Escherichia coli K-235. Biochemistry. 1967 Aug;6(8):2363–2372. doi: 10.1021/bi00860a011. [DOI] [PubMed] [Google Scholar]
  23. Millar S. J., Goldstein E. G., Levine M. J., Hausmann E. Modulation of bone metabolism by two chemically distinct lipopolysaccharide fractions from Bacteroides gingivalis. Infect Immun. 1986 Jan;51(1):302–306. doi: 10.1128/iai.51.1.302-306.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morrison D. C., Ryan J. L. Endotoxins and disease mechanisms. Annu Rev Med. 1987;38:417–432. doi: 10.1146/annurev.me.38.020187.002221. [DOI] [PubMed] [Google Scholar]
  25. Nair B. C., Mayberry W. R., Dziak R., Chen P. B., Levine M. J., Hausmann E. Biological effects of a purified lipopolysaccharide from Bacteroides gingivalis. J Periodontal Res. 1983 Jan;18(1):40–49. doi: 10.1111/j.1600-0765.1983.tb00333.x. [DOI] [PubMed] [Google Scholar]
  26. Nambu M., Morita M., Watanabe H., Uenoyama Y., Kim K. M., Tanaka M., Iwai Y., Kimata H., Mayumi M., Mikawa H. Regulation of Fc gamma receptor expression and phagocytosis of a human monoblast cell line U937. Participation of cAMP and protein kinase C in the effects of IFN-gamma and phorbol ester. J Immunol. 1989 Dec 15;143(12):4158–4165. [PubMed] [Google Scholar]
  27. Offenbacher S., Heasman P. A., Collins J. G. Modulation of host PGE2 secretion as a determinant of periodontal disease expression. J Periodontol. 1993 May;64(5 Suppl):432–444. doi: 10.1902/jop.1993.64.5s.432. [DOI] [PubMed] [Google Scholar]
  28. Ogawa T. Immunobiological properties of chemically defined lipid A from lipopolysaccharide of Porphyromonas (Bacteroides) gingivalis. Eur J Biochem. 1994 Feb 1;219(3):737–742. doi: 10.1111/j.1432-1033.1994.tb18552.x. [DOI] [PubMed] [Google Scholar]
  29. Page R. C. The role of inflammatory mediators in the pathogenesis of periodontal disease. J Periodontal Res. 1991 May;26(3 Pt 2):230–242. doi: 10.1111/j.1600-0765.1991.tb01649.x. [DOI] [PubMed] [Google Scholar]
  30. Pedrinaci S., Huelin C., Patarroyo M., Ruiz-Cabello F., Garrido F. Studies on CD11a and CD18 molecules with two new monoclonal antibodies: differential myelomonocytic antigen expression of PMA treated HL60 and U937 cell lines. Hybridoma. 1989 Feb;8(1):13–23. doi: 10.1089/hyb.1989.8.13. [DOI] [PubMed] [Google Scholar]
  31. Radzun H. J., Parwaresch M. R., Sundström C., Nilsson K., Eissner M. Monocytic origin of the human hematopoietic cell line U-937 and its convertibility to macrophages evidenced by isoenzyme mapping. Int J Cancer. 1983 Feb 15;31(2):181–186. doi: 10.1002/ijc.2910310208. [DOI] [PubMed] [Google Scholar]
  32. Redman T. K., Harmon C. C., Michalek S. M. Oral immunization with recombinant Salmonella typhimurium expressing surface protein antigen A (SpaA) of Streptococcus sobrinus: effects of the Salmonella virulence plasmid on the induction of protective and sustained humoral responses in rats. Vaccine. 1996 Jun;14(9):868–878. doi: 10.1016/0264-410x(96)00013-8. [DOI] [PubMed] [Google Scholar]
  33. Shapira L., Takashiba S., Amar S., Van Dyke T. E. Porphyromonas gingivalis lipopolysaccharide stimulation of human monocytes: dependence on serum and CD14 receptor. Oral Microbiol Immunol. 1994 Apr;9(2):112–117. doi: 10.1111/j.1399-302x.1994.tb00044.x. [DOI] [PubMed] [Google Scholar]
  34. Socransky S. S., Haffajee A. D. The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol. 1992 Apr;63(4 Suppl):322–331. doi: 10.1902/jop.1992.63.4s.322. [DOI] [PubMed] [Google Scholar]
  35. Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
  36. Ulevitch R. J., Tobias P. S. Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin. Annu Rev Immunol. 1995;13:437–457. doi: 10.1146/annurev.iy.13.040195.002253. [DOI] [PubMed] [Google Scholar]
  37. Wahl L. M., Katona I. M., Wilder R. L., Winter C. C., Haraoui B., Scher I., Wahl S. M. Isolation of human mononuclear cell subsets by counterflow centrifugal elutriation (CCE). I. Characterization of B-lymphocyte-, T-lymphocyte-, and monocyte-enriched fractions by flow cytometric analysis. Cell Immunol. 1984 May;85(2):373–383. doi: 10.1016/0008-8749(84)90251-x. [DOI] [PubMed] [Google Scholar]
  38. Wahl S. M., Katona I. M., Stadler B. M., Wilder R. L., Helsel W. E., Wahl L. M. Isolation of human mononuclear cell subsets by counterflow centrifugal elutriation (CCE). II. Functional properties of B-lymphocyte-, T-lymphocyte-, and monocyte-enriched fractions. Cell Immunol. 1984 May;85(2):384–395. doi: 10.1016/0008-8749(84)90252-1. [DOI] [PubMed] [Google Scholar]
  39. Williams R. C. Periodontal disease. N Engl J Med. 1990 Feb 8;322(6):373–382. doi: 10.1056/NEJM199002083220606. [DOI] [PubMed] [Google Scholar]

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