Project/Area Number |
15590489
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Laboratory medicine
|
Research Institution | Okayama University |
Principal Investigator |
KATAOKA Mikio Okayama University, Medical School, Professor, 医学部, 教授 (50177391)
|
Co-Investigator(Kenkyū-buntansha) |
ISHIKAWA Tetsuya Okayama University, Medical School, Associate Professor, 医学部, 助教授 (90221754)
倉園 久生 岡山大学, 医学部, 教授 (90186487)
柴倉 美砂子 岡山大学, 医学部, 助手 (30314694)
|
Project Period (FY) |
2003 – 2005
|
Project Status |
Completed (Fiscal Year 2005)
|
Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
|
Keywords | Sarcoidosis / Propionibacterium acnes / Propionibacterium granulosum / Bronchoalveolar lavage / Real-time PCR / Propoinibacterium acnes / リアルタイム PCR |
Research Abstract |
Aim : Despite the extensive investigations into the pathogenesis of sarcoidosis, its etiology is still unknown. We have reported alveolar lymphocytes from patients with sarcoidosis are sensitized by Propionibacterium acnes (P.acnes) and the proliferation and IL-2 production in these cells were seen when these cells were stimulated by P.acnes. Recently, Eishi and his colleague have reported that many genomes of P.acnes and Propionibacterium granulosum (P.granulosum) were detected in lymph nodes from sarcoidosis patients. In this study, we investigated the quantitative analysis of Propionibacterial DNA in bronchoalveolar lavage (BAL) cells in patients with sarcoidosis to clarify the role of Propionibacteria in the pathogenesis of sarcoidosis. Moreover, we investigated the serum level of antibody. Subjects and Methods : Bronchoalveolar lavage cells were obtained from 42 patients with sarcoidosis and from those with 30 non-sarcoid lung diseases. BAL cells were undergone assays of real-time
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PCR for Propionibacterial DNA in Light Cycler apparatus using Taq Man probes and primers to amplify 16S rRNA of Propionibacteria. DNA was extracted using QlAamp DNA Mini Kit and the concentration of DNA was measured in the spectrophotometer. The number of bacterial genomes in 500 ng of total DNA were estimated with an internal standard curve for each species. The serum level of antibody against P.acnes was estimated using whole bacterial ELISA method. The titer of antibody was shown as optical density. Results : The number of genomes of P.acnes in BAL cells in patients with sarcoidosis are higher than in those with non-sarcoid lung diseases. However, there was no difference in the number of genomes of P.granulosum in BAL cells between sarcoidosis patients and non-sarcoidosis patients. There are clear relationship between the number of genomes of P.acnes in BAL cells and serum angiotensin converting enzyme level and proportion of macrophage and lymphocyte in BAL fluids in patients with sarcoidosis. The antibody level against P.acnes in patients with sarcoidosis was significantly higher than in those with non-sarcoid lung diseases. Conclusion : These results suggest that P.acnes might be involved in the pathogenesis of sarcoidosis. Less
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