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Analysis of the interaction between periodontopathic bacteria and junctional epithelial cells using in situ (RT-) PCR

Research Project

Project/Area Number 10671968
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Periodontal dentistry
Research InstitutionThe University of Tokushima

Principal Investigator

NAKAE Hideaki  Tokushima Univ., Conserv. Dent., Assistant professor, 歯学部・附属病院, 講師 (30227730)

Co-Investigator(Kenkyū-buntansha) KAWASAKI Akiko  Tokushima Univ., Conserv. Dent., Instructor, 歯学部, 助手 (60294708)
YUMOTO Hiromichi  Tokushima Univ., Conserv. Dent., Instructor, 歯学部, 助手 (60284303)
YOSHIYAMA Masahiro  Tokushima Univ., Conserv. Dent., Assistant professor, 歯学部・附属病院, 講師 (10201071)
SHIMIZU Hirotoshi  Tokushima Univ., Conserv. Dent., Instructor, 歯学部, 助手 (70294709)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsPeriodontopathic bacteria / in situ-PCR / RT-PCR / Junctional epithelium / IL-8 / ICAM-1 / MAP kinase / in situ PCR / in situ / 免疫染色 / 歯周ポケット / 侵入
Research Abstract

We examined the interaction between periodontopathic bacteria and periodontal junctional epithelial cells. In RT-PCR, Significant IL-8, IL-6, PGHS-2 mRNA levels were induced from epithelial cells in response to exposure to periodontopahtic Eiklenella corrodens . Interestingly, After incubation with E. corrodens supernatant, KB cells showed marked increase in the levels of IL-8, IL-6, PGHS -2 mRNA. These results imply that the direct contact of E. corrodens with oral epithelial cells is not necessarily required for the sirumulation pf IL-6, IL-8, and Prostaglamdin E2. We suggest that E. corrodens induces the epithelial cells to secrete proinflammatory cyotkines which serve as an early signaling system to host immune and inflammatory cells in underlying conective tissue.
To elucidate the mechanism of the transmigration neutrophils through the epithelium, we investigated the expression of adhesion molecules on epithelial cells in response to E. corrodens attachment. In RT-PCR, ICAM-1 mRNA … More levels were significantly increased within 1h in response to exposure to E. corrodeens and continued over the 12h period of study. Furthermore, inhibition of the attachment of E. corrodens to KB cells with N-acetyl-D-galactosamine (GalNAc) decreased the expression of ICAM-1 and purified adhesin of E. corrodens (GalNAc specific lectin like substance, EcLS) increased the ICAM-1 expression on epithelial cells. These findings show that EcLS plays the key role in ICAM-1 expression on gingival epithelial cells.
To clarify the signal transduction in KB cells after infection of E. corrodens, we examined components that were phosphrylated by the attachment of E. corrodens to epithelial cells. In western blot analysis, phosphorylated p42/44MAK kinase was detected but phosphorylated EGF receptor was not detected. In histoimmunologycal study, phosphorylated p42/44MAP kinase were detected in epithelium but not connective tissues. Furthermore, inhibitors of MAP kinase blocked the expression of ICAM-1 and IL-8. In summary, secreted proteins from E. corrodens and EcLS can interact with epithelial cells mediating with p42/44 MAP kinase. Less

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] 藤中恵子 他: "ヒト感染根管における侵入細菌の局在に関する免疫組織学解析"日本歯科保存学雑誌. 43(印刷中). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松尾敬志 他: "歯科領域での抗菌剤の投与"医薬ジャーナル. 35. 1791-1798 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Yumoto et al.: "lnterleukin-6 (IL-6) and IL-8 are induced in human oral cpithelial cells in response to exposure to periodontopathic Eikenella corrodens"Infect.Immun.. 67,1. 384-394 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 中江英明 他: "歯学生のための歯内療法学"歯の外傷の治療法(印刷中). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Fujinaka K, et al.: "Immunohistological studies on the localization of selected bacteria in infected root canal systems"Jpn J Conserv Dent. (in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Matsuo T, et al.: "Application of antimicrobial agents in dental field"Medicine and Drug Journal. 35. 1791-1798 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yumoto H, et al.: "Interleukin-6 (IL-6) and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopathic Eikenella corrodens"Infect. Immun.. 67. 384-394 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakae H, et al.: "Treatment of injured teeth"Ishiyaku Publisher In Endodontics for dental student (in press). 28 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 藤中 恵子 他: "ヒト感染根管における侵入細菌の局在に関する免疫組織学的解析"日本歯科保存学雑誌. 43(印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 松尾 敬志 他: "歯科領域での抗菌剤の投与"医薬ジャーナル. 35. 1791-1798 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 中江 英明 他: "歯学生のための歯内療法学"歯の外傷の治療法(印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Yumoto et al.: "Interleukin-6(IL-6)and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopatjic Eikenellacorrodens" Infect.Immun.Vol.676,No.1. 384-394 (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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