| Project/Area Number |
15590312
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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 |
Human pathology
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| Research Institution | Iwate Medical University |
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Principal Investigator |
SAWAI Takashi (2005) Iwate Medical University, School of Medicine, Professor, 医学部, 教授 (00125577)
黒瀬 顕 (2003-2004) 岩手医科大学, 医学部, 講師 (70244910)
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| Co-Investigator(Kenkyū-buntansha) |
KAMATAKI Akihisa Iwate Medical University, School of Medicine, Research Associate, 医学部, 助手 (60360004)
SASAKI Nobuhito Iwate Medical University, School of Medicine, Research Associate, 医学部, 助手 (50382601)
澤井 高志 岩手医科大学, 医学部, 教授 (00125577)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | mixed connective tissue disease (MCTD) / pulmonary hypertension / anti-endothelial cell antibody (AECA) / アポトーシス / NK細胞 / サイトカイン / IL-2 / 血管内皮細胞 / MCTD / AECA / Fas |
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| Research Abstract |
The pathogenesis of pulmonary hypertension (PH) in mixed connective tissue disease (MCTD) is still unclear. Recent studies have suggested that PH in MCTD is resulted from endothelial cell injury. To reveal the pathogenic mechanism of PH in patients with MCTD, this research was focused on anti-endothelial cell antibodies (AECA) and identification of target proteins of AECA.
Cyto-ELISA was performed to detect AECA in MCTD patients with PH using micro vascular endothelial cell (HMVEC-L). To determine AECA prevalence, values higher than 2 S.D. above the mean of healthy control were considered as positive. Six of 9 MCTD patients had positive results. The titer of AECA was higher in MCTD patients with PH than without PH.
For identification of targets of AECA, proteins from HMVEC-L were separated by two-dimensional electrophoresis and transferred on PVDF membranes. Western blot analyses were performed with sera from healthy control or MCTD patients. Detected protein spots with a difference among the samples were selected and identified by peptide mass finger printing. About 20 protein spots were detected and half of these were identified.
As the results of two-dimensional western blot analyses, candidate antigens were obtained. These included proteins reported as autoantigen in various diseases. Furthermore, one of candidates is suggested to be associated with PH in patients with SSc. This raises possibility that the identified proteins include antigens of AECA, indicating that two-dimensional western blot analysis is useful technique for detection of the target antigens of AECA.
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