| Project/Area Number |
10670751
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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 |
Pediatrics
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| Research Institution | Nara Medical University |
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Principal Investigator |
SHIMA Midori Nara Medical University, Dept of Pediatrics, Associate Professor, 医学部・小児科, 助教授 (30162663)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Ichiro Nara Medical University, Dept of Pediatrics, Assistant Professor, 医学部・小児科, 講師 (00201616)
中 宏之 奈良県立医科大学, 医学部, 助手 (40281761)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | factor VIII / hemophilia A / anti factor VIII alloantibody / inhibitor / factor VIII gene / 遺伝子治療 / C2ドメイン / 同種抗体 |
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| Research Abstract |
1.Factor VIII expression : Recombinant adeno viral vector consisting of Cytomegalovirus enhancer/chiken _-actin promoter/rabbit _-globulin 3'UTR, B domain less canine factor VIII gene and rabbit _-globulin poly A was constructed. COS1, HepG2 and Hela cells were transfected and FVIII was expressed. Approximately 0.6 unit/ml and 1.0 unit/ml of factor VIII was detected by COS1 cells at day 4 and 7, respectively. Furthermore, 0.8 unit/ml of and 1.0unit/ml of factor VIII was detected by HepG2 cells at day 4 and day 7, respectively. Factor VIII was not detected by Hela cells.
2.Immunological influence : Immunological and biological characterizations of anti-factor VIII alloantibodies were analyzed. Especially, (1)IgG subclass specificity, (2)binding region of the alloantibodies, (3)inhibitory mechanism on factor VIII cofactor function were determined. We demonstrated for the first time the inhibitory effects on factor IX binding, factor VIII binding to von Willebrand factor and activated platelets and inhibitory effects on factor VIII activation by thrombin and activated factor Xa. Furthermore, we detected factor VIII/anti-factor VIII autoantibodies immune complex in plasma.
3.Interaction between factor VIII genotype of heamophilia A patient and development of anti-factor VIII alloantibodies : We found a close relationship between HLA-DR4.1, DQ4 and DQA1 and inhibitor development. Furthermore we also found that some factor VIII genotype was associated with the development of inhibitor.
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