Inhibition of tissue-reparing signaling by P.gingivalis infection

• Project/Area Number: 15390645
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Periodontal dentistry
• Research Institution: Osaka University
• Principal Investigator: OKAHASHI Nobuo Osaka University, Graduate School of Dentistry, Associate Professor, 大学院・歯学研究科, 助教授 (40150180)
• Co-Investigator(Kenkyū-buntansha): AMANO Atsuo Osaka University, Graduate School of Dentistry, Professor, 大学院・歯学研究科, 教授 (50193024) ; NAKAGAWA Ichiro Osaka University, Graduate School of Dentistry, Assistant Professor, 大学院・歯学研究科, 講師 (70294113)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥7,900,000 (Direct Cost: ¥7,900,000); Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000); Fiscal Year 2003: ¥6,000,000 (Direct Cost: ¥6,000,000)
• Keywords: periodontitis / tissue regeneration / P.gingivalis / cellular attachment / epithelial cells / periodontal ligament cells / 骨芽細胞

Research Abstract

P.gingivalis, a periodontopathic Gram-negative bacteria, is known to invade into a variety of host cells such as epithelial cells and fibroblasts. A chronic inflammation of infected periodontal tissue results in tissue destruction. In this study, we investigated the involvement of the two major virulent factors of this pathogen ; fimbriae and a protease called gingipain on the destruction and inhibition of regeneration of infected host tissues.
At first, we constructed fimbriae-deficient and gingipain-deficient mutants of P.gingivalis. Using these two mutants, we found that :
1)Both fimbriae- and gingipain-deficient mutants of P.gingivalis showed decreased invasion in human periodontal ligament cells (PDLs).
2)Infection of wild type of P.gingivalis in serum-free medium resulted in a detachment of PDLs from the culture dishes. However, gingipain-deficient mutant did not induce such detachment of PDLs.
3)Infection of wild type of P.gingivalis induced a degradation of integrin-associated signaling molecules such as FAK and paxillin. However, the degradation of FAK and paxillin was not observed during infection with gingipain-deficient mutant.
4)By using a scratch assay, we found that the infection of P.gingivalis strongly inhibited the regeneration of damaged monolayers of PDLs. The bacterial infection inhibited the cellular movement as well as the cellular growth.
5)Gingipain-deficient mutant did not show the inhibitory effect on the regeneration of the infected cellular monolayers. On the other hand, fimbriae-deficient mutant did inhibit the regeneration process.
These results strongly suggested that gingipain of P.gingivalis plays an important role in the inhibition of regeneration of the infected periodontal tissues.

Research Products

• [Journal Article] Porphyromonas gingivalis induces receptor activator of NF-kB ligand expression in osteoblasts through the activator protein-1 pathway. (2004)
  • Author(s): Okahashi N, Inaba H, Nakagawa I, Yamamura T, Kuboniwa M, Nakayama K, Hamada S, Amano A
  • Journal Title: Infection and Immunity 72 Pages: 1706-1714
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Variations of Porphyromonas gingivalis fimbriae in relation to microbial pathogenesis. (2004)
  • Author(s): Amano A, Nakagawa I, Okahashi N, Hamada S
  • Journal Title: Journal of Periodontal Research 39 Pages: 136-142
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probe (2004)
  • Author(s): Kuboniwa M, Amano A, Kimura RK, Sekine S, Kato S, Yamamoto Y, Okahashi N, Iida T, Shizukuishi S
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 168-176
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Comparison of inflammatory changes caused by Porphyromonas gingivalis with distinct fimA genotypes in a mouse abscess model. (2004)
  • Author(s): Nakano K, Kuboniwa M, Nakagawa I, Yamamura Y, Nomura R, Okahashi N, Ooshima T, Amano, A
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 205-209
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effect of enamel matrix derivative on periodontal ligament cells diminished by Porphyromonas gingivalis (2004)
  • Author(s): Inaba H, Kawai S, Nakayama K, Okahashi N, Amano A
  • Journal Title: Journal of Periodontology 75 Pages: 865-872
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Autophagy defends cells against invading group A streptococci (2004)
  • Author(s): Nakagawa I, Amano A, Mizushima N, Yamamoto A, Yamaguchi H, Kamimoto T, Nara A, Funao J, Nakata M, Tsuda K, Hamada S, Yoshimori T
  • Journal Title: Science 306 Pages: 1037-1040
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Porphyromonas gingivalis induces receptor activator of NF-kB ligand expression in osteoblasts throuhg the activator protein-1 pathway. (2004)
  • Author(s): Okahashi, N., Inaba, H., Nakagawa, I., Yamamura, T., Kuboniwa, M., Nakayama, K., Hamada, S., Amano, A.
  • Journal Title: Infection and Immunity 72 Pages: 1706-1714
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Variations of Porphyromonas gingivalis fimbriae in relation to microbial pathogenesis. (2004)
  • Author(s): Amano, A., Nakagawa, I.Okahashi, N., Hamada, S.
  • Journal Title: Journal of Periodontal Research 39 Pages: 136-142
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Quantitative detection of periodontal pathogenesis using real-time polymerase chain reaction with TaqMan probe. (2004)
  • Author(s): Kuboniwa, M., Amano, A., Kimura, R.K., Sekine, S., Kato, S., Yamamoto, Y., Okahashi, N., Iida, T., Shizukuishi, S.
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 168-176
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Comparison of inflammatory changes caused by Porphyromonas gingivalis with distinct fimA genotypes in a mouse abscess model. (2004)
  • Author(s): Nakano, K., Kuboniwa, M., Nakagawa, I., Yamamura, T., Nomura, R., Okahashi, N., Ooshima, T., Amano, A.
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 205-209
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effect of enamel matrix derivative on periodontal ligament cells diminished by Porphyromonas gingivalis. (2004)
  • Author(s): Inaba, H., Kawai, S., Nakayama, K., Okahashi, N., Amano, A.
  • Journal Title: Journal of Periodontology 75 Pages: 865-872
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Autophagy defends cells against invading group A streptococcus. (2004)
  • Author(s): Nakagawa, I., Amano, A., Mizushima, N., Yamamoto, A., Yamaguchi, H., Kamimoto, T., Nara, A., Funao, J., Nakata, M., Tsuda, K., Hamada, S., Yoshimori, T.
  • Journal Title: Science 306 Pages: 1037-1040
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Porphyromonas gingivalis induces receptor activator of NF-kB ligand expression in osteoblasts through the activator protein a pathway. (2004)
  • Author(s): Okahashi N, Inaba H, Nakagawa I, Yamamura T, Kuboniwa M, Nakayama K, Hamada S, Amano A
  • Journal Title: Infection and Immunity 72 Pages: 1706-1714
• [Journal Article] Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probe (2004)
  • Author(s): Kuboniwa M, Amano A, Kimura RK, Sekine S, Kato S, Yamamoto Y, Okahashi N, Iida T.
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 168-176
• [Journal Article] Comparison of inflammatory changes caused by Porphyromonas gingivalis with distinct fimA genotypes in a mouse abscess model. (2004)
  • Author(s): Nakano K, Kuboniwa M, Nakagawa I, Yamamura Y, Nomura R, Okahashi N, Ooshima T, Amano A
  • Journal Title: Oral Microbiology and Immunology 19 Pages: 205-209
• [Journal Article] Autophagy defends cells against invading group A streoptococc (2004)
  • Author(s): Nakagawa I, Amano A, Mizushima N, Yamamoto A, Yamaguchi H, Kamimoto T, Nara A, Funao J, Nakata M, Tsuda K, Hamada S, Yoshimori T
  • Journal Title: Science 306 Pages: 1037-1040
• [Publications] Itoh K, Udagawa N, Kobayashi K, Suda K, Li X, Takami M, Okahashi N, Nishihara T, Takahashi N.: "Lipopolysaccharide promotes the survival of osteoclasts via Toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophages"Journal of Immunology. 170. 3688-3695 (2003)
• [Publications] Tani H, Limn CK, Yap CC, Onishi M, Nozaki M, Nishimune Y, Okahashi N, Kitagawa Y, Watanabe R, Mochizuki R, Moriishi K, Matsuura Y.: "In vitro and in vivo gene delivery by recombinant baculoviruses"Journal of Virology. 77. 9799-9808 (2003)
• [Publications] Okahashi N, Sakurai A, Nakagawa I, Fujiwara T, Kawabata S, Amano A, Hamada S.: "Infection by Streptococcus pyogenes induces the receptor activator of NF-kB ligand expression in mouse osteoblastic cells"Infection and Immunity. 71. 948-955 (2003)
• [Publications] Nakagawa I, Kurokawa K, Yamashita A, Nakata M, Tomiyasu Y, Okahashi N, Kawabata S, Yamazaki K, Shiba T, Yasunaga T, Hayashi H, Hattori M, Hamada S.: "Genomic sequence of an M3 strain of Streptococcus pyogenes reveals a large scale genomic rearrangement in invasive strains and new insights into phage evolution"Genome Research. 13. 1042-1055 (2003)
• [Publications] Sakurai A, Okahashi N, Nakagawa I, Kawabata S, Amano A. Ooshima T, Hamada S.: "Streptococcus pyogenes infection induces septic arthritis with increased production of receptor activator of NF-kB ligand"Infection and Immunity. 71. 6019-6026 (2003)
• [Publications] Nakano K, Nomura R, Nakagawa I, Hamada S, Ooshima T.: "Determination of Streptococcus mutans with a cell wall polysaccharide specific to a new serotype k, in the human oral cavity"Journal of Clinical Microbiology. 42. 198-202 (2004)