| Project/Area Number |
12670440
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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 |
内科学一般
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| Research Institution | TOKYO WOMEN'S MEDICAL UNIVERSITY |
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Principal Investigator |
HARIGAI Masayoshi Tokyo Women's Medical University, Institute of Rheumatology, Assistant Professor, 医学部, 講師 (20238207)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Michi Tokyo Women's Medical University, Institute of Rheumatology, Research Fellow, 医学部, 助手 (50277141)
KAWAGUCHI Yasushi Tokyo Women's Medical University, Institute of Rheumatology, Research Fellow, 医学部, 助手 (90297549)
HARA Masako Tokyo Women's Medical University, Institute of Rheumatology, Professor, 医学部, 教授 (80090009)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | rheumatoid arthritis / tumor necrosis factor-α / tristetraprolin / gene therapy / tristetraproline / RNA結合蛋白 / AU rich / 慢性関節リウマチ / RNA結合蛋白質 |
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| Research Abstract |
Background : TNF-α plays a central role in the immunopathogenesis of rheumatoid arthritis (RA). Expression of TNF-α is tightly regulated by a series of regulatory mechanisms in vivo. These include transcriptional, translational and post-translational regulation. Tristetraprolin (TTP) is a recently described RNA binding protein which is responsible for degradation of TNF-α messenger RNA (mRNA). It binds to 3'-untranslated region of TNF-α mRNA and accelerates degradation of the transcript. In this study, we explored the expression of TTP in synovial tissue from patients with RA and prepared adenovirus for inducible expression of TTP. Methods : Expression of TTP in synovial tissue of patients with RA or osteoarthritis (OA) was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. Adenovirus was made using in vitro ligation method. Messenger RNA was quantified using ABI Prism 7900. Results : TTP mRNA was detected in 5 out of 6
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synovial tissue samples from patients with RA using RT-PCR. TTP protein was detected in all synovial tissue samples using Western blotting. Immunohistochemistry revealed expression of TTP in the lining layer of synovial tissue as well as around micro-vessels in the sublining region of synovial tissue from patients with RA. Synovial tissue from patients with OA did not express TTP. The expression plasmids for human and mice TTP was prepared and transfected into COS7 cells. These plasmid successfully induced expression of human or mice TTP in these cells. Human TTP cDNA along with tetracycline responsive element was introduced into adenovirus (human TTP adenovirus). When COS7 cells were co-infected with human TTP adenovirus and tetracycline-controlled transactivator virus, strong expression of human TTP was induced. Conclusion : TTP is strongly expressed in synovial tissue from patients with RA, but not from OA. Adenovirus which can induce expression of human TTP was prepared. Investigation of the effects of human TTP adenovirus gene therapy against experimental arthritides should be determined in future. Less
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