Studies on structure and function of factor VIII and Von willebrand factor complex in hemophiliacs
Project/Area Number |
03454507
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Research Category |
Grant-in-Aid for General Scientific Research (B)
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Allocation Type | Single-year Grants |
Research Field |
Laboratory medicine
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Research Institution | Tokyo Medical College |
Principal Investigator |
FUKUTAKE Katsuyuki Tokyo Medical College, Clinical Pathology, professor, 医学部, 教授 (40192306)
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Project Period (FY) |
1991 – 1993
|
Project Status |
Completed (Fiscal Year 1993)
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Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥1,100,000 (Direct Cost: ¥1,100,000)
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Keywords | factor VIII / von Willebrand factor / homophilia / von Willebrand disease / von Willebrand factor gene / point mutation / PCR / ポリモルフィズム / 第VIII因子 / 時間分解蛍光光度計 |
Research Abstract |
Hitherto abnormalities of blood coagulation factor VIII (F.VIII) and von willebrand factor (vWF) have been recognized hemophilia A and von Willebrand disease. However, paralleling progress in the analysis of the proteins of these coagulation factors it has become clear that these abnormalities are not lomited merely to congenital protein deficiencies but are also seen in a variety of complex disease states associated with various protein structural abnomalities. By studying antibodies developing to F.VIII in humans and analyzing the epitope on the protein of F.VIII we investigated the relation between F.VIII and vWF.It was found thaat the epitope of the antibodies recgnizing the light chain is present on C2 domein, and that these antibodies work more strongly against F.VIII alone than aagainst FVIII-vWF comples, suggesting that near this epitope changes in complex formation are induced. Also, with regard to vWF, type II von willebrand disease patients were analyzed regarding exon 28 which is a site expresing important fanctions. Tow types of polymorphism were discovered, and were confirmed to be useful in the diagnosis of this disease. Regarding von Willebrand factor gene in 19 von Willebrand disease patients, analysis was performed by amplification by PCR, restriction enzyme MspI digestion and electrophoresis, AluI digestion and Mutation Detection Enhancement (MDE) gels electrophoresis. In 5 cases abnormal patterns were obtained by analysis with restriction enzyme MspI digestion of exon 28 and MDE gel electrophoresis, hetero-duplex bands were detected in 4 cases. From this study, 6 types of point mutation were discovered in exon 28 of vWF gene, while in other exons any mutations were not discovered. On the other hand, in 40 hemophilia A patients missense mutations in codon 854 of exon 20 showing the Normandy variant were searched for, although not a single case was detected.
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Report
(4 results)
Research Products
(12 results)