Molecular and genetic analysis of polymorphonuclear luekocytes in early onset periodontitis patients
| Project/Area Number |
06454536
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Conservative dentistry
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| Research Institution | Niigata University |
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Principal Investigator |
YOSHIE Hiromasa Niigata University School of Dentistry, Associate professor, 歯学部, 助教授 (20143787)
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| Co-Investigator(Kenkyū-buntansha) |
NOMURA Takashi Niigata University School of Dentistry, Research instructor, 歯学部・付属病院, 助手 (70251840)
MATSUKI Yutaka Niigata University School of Dentistry, Research instructor, 歯学部, 助手 (70242435)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1994: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | early onset periodontitis / polymorphonuclear leukocyte / Fcgamma receptor / complement receptor / phagocytosis / mRNA / gingival crevicular fluid / 貪食能 / Fcレセプター |
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| Research Abstract |
I.Immunoglobulin G type II and III receptors (FcgammaRII and FcgammaRIII) are essential for polymorphonuclear leukocytic (PMNs) phagocytosis. To determine whether this receptor downregulation may contribute to the periodontal host defense borne by PMNs, we examined the correlation between FcgammaRII and FcgammaRIII expressions and the phagocytic capacity of GCF-PMNs. In order to verify at which level of cellular events the loss of FcgammaR occurs, we quantified mRNA levels to assess a de novo synthesis of these receptors. GCF was collected from 21 patients with adult periodontitis by gingival crevicular washing. Autologous peripheral blood (PB) PMNs served as control. Surface expressions of FcgammaRs and phagocytic capacity via FcgammaRs were analyzed by flow cytometry. The difference in FcgammaR mRNA levels between GCF- and PB-PMNs was assessed by reverse transcription-polymerase chain reaction (RT-PCR). The amplified products were visualized by agarose gel electrophoresis and the end
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product yields were quantified by computerized image-analysis. Both FcgammaRII and FcgammaRIII expressions and phagocytic capacity on GCF-PMNs were significantly lower than those on PB-PMNs (p<0.001). The mRNA level of FcgammaRIII of GCF-PMNs was significantly lower than that of PB-PMNs. Thus, GCF-PMNs are characterized by the decreased surface expressions and mRNA levels of FcgammaRs, and the impaired phagocytosis.
II.In this study we attempted to determine the mRNA levels of complement receptor type1 and 3 (CR1, CR3) on PMNs in GCF by using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). GCF samples were obtained from 11 adult periodontitis patients by gingival crevicular washing, and venipunctured PB was used as a control. RT-PCR analysis was performed using the primer sets for CR1, CR3 and beta-actin. Digoxigenin-labelled RNA probes were synthesized from RT-PCR products for ISH.Both CR1 and CR3 mRNA levels relative to beta-actin were significantly lower in GCF-PMNs than in PB-PMNs. In ISH,a greater majority of PB-PMNs showed positive CR1 and CR3 mRNA expressions, while only a few PMNs showed positive signals in GCF.Our data in this study suggest that increased expressions of CR on the PMN cell surface appear to be unrelated to de novo synthesis. Less
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Report
(4 results)
Research Products
(9 results)
