1999 Fiscal Year Final Research Report Summary
Analysis of the interaction between periodontopathic bacteria and junctional epithelial cells using in situ (RT-) PCR
| Project/Area Number |
10671968
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Periodontal dentistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
NAKAE Hideaki Tokushima Univ., Conserv. Dent., Assistant professor, 歯学部・附属病院, 講師 (30227730)
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| 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)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Periodontopathic bacteria / in situ-PCR / RT-PCR / Junctional epithelium / IL-8 / ICAM-1 / MAP kinase |
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| 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
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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
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