Skip to main content
SpringerLink
Log in

Activation of Transcription Factors and IL-8 Expression in Neutrophils Stimulated with Lipopolysaccharide from Porphyromonas gingivalis

  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

DNA binding activity of NF-κB and AP-1 were examined in neutrophils stimulated with LPS purified from P. gingivalis, a major pathogenic bacteria of periodontitis lesion. Porphyromonas gingivalis LPS enhanced the activity reaching a peak at a concentration of 500 ng/ml in the absence of serum. The NF-κB activation stimulated with 10 ng/ml of P. gingivalis LPS was suppressed approximately 44% by treatment of neutrophils with anti-CD14 antibody under the presence of serum. Increase in the steady-state IL-8 mRNA level was concomitantly observed by stimulation of neutrophils with 500 ng/ml of P. gingivalis LPS under the absence of serum. These results indicate that P. gingivalis LPS activates NF-κB and AP-1 in both serum-dependent and -independent manners, followed by increased IL-8 transcription in neutrophils, and suggested a role for P. gingivalis LPS in IL-8 synthesis by neutrophils in inflamed gingiva and GCF.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. Dubravec, D. B., D. R. Spriggs, J. A. Mannick, and M. L. Rodrick. 1990. Circulating human peripheral blood granulocytes synthesize and secrete tumor necrosis factor α. Proc. Natl. Acad. Sci. U.S.A. 87:6758–6761.

    PubMed  Google Scholar 

  2. Strieter, R. M., K. Kasahara, R. M. Allen, T. J. Standiford, M. W. Rolfe, F. S. Becker, S. W. Chensue, and S. L. Kunkel. 1992. Cytokine-induced neutrophil-derived interleukin-8. Am. J. Pathol. 141:397–407.

    PubMed  Google Scholar 

  3. Cicco, N. A., A. Lindemann, J. Content, P. Vandenbussche, M. LÜbbert, J. Gauss, R. Mertelsmann, and F. Herrmann. 1990. Inducible production of interleukin-6 by human polymorphonuclear neutrophils: Role of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha. Blood 75:2049–2052.

    PubMed  Google Scholar 

  4. Lloyd, A. R., and J. J. Oppenheim. 1992. Poly's lament: the neglected role of the polymorphonuclear neutrophil in the afferent limb of the immune response. Immunol. Today 13:169–172.

    Article  PubMed  Google Scholar 

  5. Liu, J. H., and J. Y. Djeu. 1995. Role of cytokines in neutrophil functions. In Human Cytokines: Their Role in Disease and Therapy. (B. B. Aggarwal and R. K. Puri, eds.), Blackwell Science, Cambridge. 71–86.

    Google Scholar 

  6. Sugita, N., T. Suzuki, H. Yoshie, N. Yoshida, M. Adachi, and K. Hara. 1993. Differential expression of CR3, FeeRII and Fc γ RIII on polymorphonuclear leukocytes in gingival crevicular fluid. J. Periodont. Res. 28:363–372.

    PubMed  Google Scholar 

  7. Aida, Y., T. Kukita, H. Takada, K. Maeda, and M. J. Pabst. 1995. Lipopolysaccharides from periodontal pathogens prime neutrophils for enhanced respiratory burst: differential effect of a synthetic lipid a precursor IVA (LA-14-PP). J. Periodont. Res. 30:116–123.

    PubMed  Google Scholar 

  8. Wilson, M. 1993. Biological activities of lipopolysaccharide and endotoxin. In Biology of the species Porphyromonas gingivalis (H. N. Shah, D. Mayrand and R. J. Genco, eds.), CRC Press, Boca Raton, Florida 171–197.

    Google Scholar 

  9. Shapira, L., S. Takashiba, C. Champagne, S. Amar and T. E. Van Dyke. 1994. Involvement of protein kinase C and protein tyrosine kinase in lipopolysaccharide-induced TNF-α and IL-1β production by human monocytes. J. Immunol. 153:1818–1824.

    PubMed  Google Scholar 

  10. Koga, T., T. Nishihara, T. Fujiwara, T. Nisizawa, N. Okahashi, T. Noguchi, and S. Hamada. 1985. Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli. Infect. Immun. 47:638–647.

    PubMed  Google Scholar 

  11. McDonald, P. P., A. Bald and M. A. Cassatella. 1997. Activation of the NF-κB pathway by inflammatory stimuli in human neutrophils. Blood 89:3421–3433.

    PubMed  Google Scholar 

  12. Mukaida, N., S. Okamoto, Y. Ishikawa, and K. Matsushima. 1994. Molecular mechanism of interleukin-8 gene expression. J. Leukoc. Biol. 56:554–558.

    PubMed  Google Scholar 

  13. Tamura, H., S. Tanaka, T. Obayashi, M. Yoshida, and T. Kawai. 1992. A new sensitive microplate assay of plasma endotoxin. J. Clin. Lab. Anal. 6:232–238.

    PubMed  Google Scholar 

  14. English, D., and B. R. Andersen. 1974. Single-step separation of red blood cells, granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J. Immunol. Methods 5:249–252.

    Article  PubMed  Google Scholar 

  15. Meda, L., S. Gasperini, M. Ceska, and M. A. Cassatella. 1994. Modulation of proinflammatory cytokine release from human polymorphonuclear leukocytes by gamma interferon. Cell. Immunol. 157:448–461.

    Article  PubMed  Google Scholar 

  16. Shapira, L., S. Takashiba, S. Amar and T. E. Van Dyke. 1994. Porphyromonas gingivalis lipopolysaccharide stimulation of human monocytes: dependence on serum and CD14 receptor. Oral Microbiol. Immunol. 9:112–117.

    PubMed  Google Scholar 

  17. Osborn, L., S. Kunkel, and G. J. Nabel. 1989. Tumor necrosis factor α and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor κB. Proc. Natl. Acad. Sci. U.S.A. 84:2336–2340.

    Google Scholar 

  18. Thieblemont, N., N. Haeffner-Cavaillon, A. Haeffner, B. Cholley, L. Weiss, and M. D. Kazatchkine. 1995. Triggering of complement receptors CR1 (CD35) and CR3 (CD11b/CD18) induces nuclear translocation of NF-κB (p50/p65) in human monocytes and enhances viral replication of HIV-infected monocytic cells. J. Immunol. 155:4861–4867.

    PubMed  Google Scholar 

  19. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principal of protein-dye binding. Anal. Biochem. 72:248–254.

    Article  PubMed  Google Scholar 

  20. Adcock, I. M., S. J. Lane, C. R. Brown, M. J. Peters, T. H. Lee, and P. J. Barnes. 1995. Differences in binding of glucocorticoid receptor to DNA in steroid-resistant asthma. J. Immunol. 154:3500–3505.

    PubMed  Google Scholar 

  21. Nomura, T., T. Takahashi and K. Hara. 1993. Expression of TIMP-1, TIMP-2 and collagenase mRNA in periodontitis-affected human gingival tissue. J. Periodont. Res. 28:354–362.

    PubMed  Google Scholar 

  22. Sugita, N., Y. Matsuki, H. Yoshie, and K. Hara. 1997. Characterization of complement receptor type 1 and type 3 mRNA expression on polymorphonuclear leucocytes (neutrophils) in gingival crevicular fluid from periodontitis-affected patients. Arch. Oral Biol. 42:113–120.

    Article  PubMed  Google Scholar 

  23. Chelly, J., J. C. Kaplan, P. Maire, S. Gautron, and A. Kahn. 1988. Transcription of the dytrophin gene in human muscle and non-muscle tissues. Nature 333:858–860.

    Article  PubMed  Google Scholar 

  24. Tokoro, Y., T. Yamamoto, and K. Hara. 1996. IL-1β mRNA as the predominant inflammatory cytokine transcript: correlation with inflammatory cell infiltration into human gingiva. J. Oral Pathol. Med. 25:225–231.

    PubMed  Google Scholar 

  25. Ishikawa, Y., N. Mukaida, K. Kuno, N. Rice, S. Okamoto, and K. Matsushima. 1995. Establishment of lipopolysaccharide-dependent nuclear factor κB activation in a cell-free system. J. Biol. Chem. 270:4158–4164.

    Article  PubMed  Google Scholar 

  26. Ding, A., S. Hwang, H. M. Lander, and Q. W. Xie. 1995. Macrophages derived from C3H/HeJ (Lpsd) mice respond to bacterial lipopolysaccharide by activating NF-κB. J. Leukoc. Biol. 57:174–179.

    PubMed  Google Scholar 

  27. Matsusaka, T., K. Fujikawa, Y. Nishio, N. Mukaida, K. Matsushima, T. Kishimoto, and S. Akira. 1993. Transcription factors NF-IL6 and NF-κB synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc. Natl. Acad. Sci. U.S.A. 90:10193–10197.

    PubMed  Google Scholar 

  28. Baeuerle, P. A., and T. Henkel. 1994. Function and activation of NF-κB in the immune system. Annu. Rev. Immunol. 12:141–179.

    PubMed  Google Scholar 

  29. Fujishima, S., A. R. Hoffman, T. Vu, K. J. Kim, H. Zheng, D. Daniel, Y. Kim, E. F. Wallace, J. W. Larrick, and T. A. Raffin. 1993. Regulation of neutrophil interleukin 8 gene expression and protein secretion by LPS, TNF-α, and IL-1β. J. Cell. Physiol. 154:478–485.

    PubMed  Google Scholar 

  30. Wright, S. D., R. A. Ramos, P. S. Tobias, R. J. Ulevitch, and J. C. Mathison. 1990. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science 249:1431–1433.

    PubMed  Google Scholar 

  31. Schumann, R. R., S. R. Leong, G. W. Flaggs, P. W. Gray, S. D. Wright, J. C. Mathison, P. S. Tobias, and R. J. Ulevitch. 1990. Structure and function of lipopolysaccharide binding protein. Science 249:1429–1431.

    PubMed  Google Scholar 

  32. Simmons, D. L., S. Tan, D. G. Tenen, A. Nicholson-Weller and B. Seed. 1989. Monocyte antigen CD14 is a phospholipid anchored membrane protein. Blood 73:284–289.

    PubMed  Google Scholar 

  33. Haziot, A., B. Z. Tsuberi, and S. M. Goyert. 1993. Neutrophil CD14: Biochemical properties and role in the secretion of tumor necrosis factor-α in response to lipopolysaccharide. J. Immunol. 150:5556–5565.

    PubMed  Google Scholar 

  34. Mukaida, N., Y. Ishikawa, N. Ikeda, N. Fujioka, S. Watanabe, K. Kuno, and K. Matsushima. 1996. Novel insight into molecular mechanism of endotoxin shock: biochemical analysis of LPS receptor signaling in a cell-free system targeting NF-κB and regulation of cytokine production/action through β2 integrin in vivo. J. Leukoc. Biol. 59:145–151.

    PubMed  Google Scholar 

  35. Shapira, L., C. Champagne, B. Gordon, S. Amar, and T. E. Van Dyke. 1995. Lipopolysaccharide priming of superoxide release by human neutrophils: Role of membrane CD14 and serum LPS binding protein. Inflammation 19:289–295.

    PubMed  Google Scholar 

  36. Hailman, E., T. Vasselon, M. Kelley, L. A. Busse, M. C. T. Hu, H. S. Lichenstein, P. A. Detmers, and S. D. Wright. 1996. Stimulation of macrophages and neutrophils by complexes of lipopolysaccharide and soluble CD14. J. Immunol. 156:4384–4390.

    PubMed  Google Scholar 

  37. Wright, S. D., S. M. Levin, M. T. C. Jong, Z. Chad, and L. G. Kabbash. 1989. CR3 (CD11b/CD18) expresses one binding site for Arg-Gly-Asp-containing peptides and a second site for bacterial lipopolysaccharide. J. Exp. Med. 169:175–183.

    Article  PubMed  Google Scholar 

  38. Nair, B. C., W. R. Mayberry, R. Dziak, P. B. Chen, M. J. Levine, and E. Hausmann. 1983. Biological effects of a purified lipopolysaccharide from Bacteroides gingivalis. J. Periodont. Res. 18:40–49.

    PubMed  Google Scholar 

  39. Harada, A., N. Sekido, T. Akahoshi, T. Wada, N. Mukaida, and K. Matsushima. 1994. Essential involvement of interleukin-8 (IL-8) in acute inflammation. J. Leukoc. Biol. 56:559–564.

    PubMed  Google Scholar 

  40. Beaulieu, A. D., and S. R. McColl. 1994. Differential expression of two major cytokines produced by neutrophils, interleukin-8 and the interleukin-1 receptor antagonist, in nuetrophils isolated from the synovial fluid and peripheral blood of patients with rheumatoid arthritis. Arthritis Rheum. 57:855–859.

    Google Scholar 

  41. Tonetti, M. S., M. A. Imboden, L. Gerber, N. P. Lang, J. Laissue, and C. Mueller. 1994. Localized expression of mRNA for phagocyte-specific chemotactic cytokines in human periodontal infections. Infect. Immun. 62:4005–4014.

    PubMed  Google Scholar 

  42. Furie, M. B., and G. J. Randolph. 1995. Chemokines and tissue injury. Am. J. Pathol. 146:1287–1301.

    PubMed  Google Scholar 

  43. Gura, T. 1996. Chemokines take center stage in inflammatory ills. Science 272:954–956.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Periodontology, Niigata University School of Dentistry, Japan

    Noriko Sugita, Asako Kimura, Yutaka Matsuki, Hiromasa Yoshie & Kohji Hara

  2. Institute of Nephrology, Niigata University School of Medicine, Japan

    Asako Kimura & Tadashi Yamamoto

Authors
  1. Noriko Sugita
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Asako Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yutaka Matsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tadashi Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hiromasa Yoshie
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kohji Hara
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sugita, N., Kimura, A., Matsuki, Y. et al. Activation of Transcription Factors and IL-8 Expression in Neutrophils Stimulated with Lipopolysaccharide from Porphyromonas gingivalis . Inflammation 22, 253–267 (1998). https://doi.org/10.1023/A:1022344031223

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022344031223

Keywords

Search

Navigation