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Study on functinal domains of von Willebrand (vW) factor in patients with vW disease.

vW 病患者血管性血友病 (vW) 因子功能域的研究。

基本信息

批准号：
01570744
负责人：
TAKAHASHI Yukihiro
金额：
$ 1.34万
依托单位：
Nara Medical University (1991)The University of Tokyo (1989-1990)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

I. The analysis of functional domains of von Willebrand factor(vWf) and the production and characterization of monoclonal antibodies (MAbs) against vWf.A. the factorVIII-binding domain: vWf-plasmin or trypsin digested fragment p-34(amino acid residue 1-298(273)) was purified using DEAE and Heparin column. 4 different MAbs against p-34 were produced by D.Meyer, INSERM 143 France, collaborated with me. Now, I'm going to characterize these antibodiesB. Platelet glycoprotein Ib(GPIb)-binding domain: Two different MAbs (NMC/vW4 from our lab. 322 from france) against vWf which inhibit the binding of vWf to GPIb were characterized, and I discovered the different functional site of GPIb-binding of vWf by two inducers, such as antibiotics ristocetin and venom factor botrocetin. and Aurin tricarboxylic acid (ATA) is known to inhibit ristocetin-induced platelet aggregation but not the other platelet aggregaters. Its capacity to abolish human vWf-platelet interaction was further in vestigated and was found that ATA which blocks the vWf/GPIb pathway by interfering with vWf and not with platelet, is a potential tool in prevention the early stages of thrombosisC. Human collagen-binding domain: One MAb(NMC/vW3) which inhibit the vWf-human typeIII fibrilar collagen binding was discovered and was shown the epitope in vWf subunit. That is, The epitope was localized between the amino acid residue 911 and 1365.II. The analysis of plasma von Willebrand factor in von Willebrand disease. I studied on the establishment of several vWf function assays and vWf protein analysis of plasma vWf in normal and several types of von Willebrand disease.A. The vWf-human collagen binding assay, using a minute plasma was established. The abnormality of the collagen- binding in type II vWd was discovered.B. The analytical method of vWf subunit in plasma vWf using a minute sample was established. Now the plasma vWf in several types of vWd is going to examine.

I.血管性血友病因子（vWf）的功能结构域分析及抗vWf. A单克隆抗体的制备和鉴定凝血因子VIII结合结构域：使用DEAE和肝素柱纯化vWF-纤溶酶或胰蛋白酶消化的片段p-34（氨基酸残基1-298（273））。4种不同的抗p-34单克隆抗体是由D.Meyer，CIM 143法国，与我合作生产的。血小板糖蛋白Ib（GPIb）结合结构域：两种不同的单克隆抗体（来自我们实验室的NMC/vW 4）。322）对vWf与GPIb结合的抑制作用进行了研究，发现了两种诱导剂（抗生素瑞斯托霉素和蛇毒因子肉毒素）对vWf与GPIb结合的不同功能位点。已知金精三羧酸（ATA）抑制瑞斯托菌素诱导的血小板聚集，但不抑制其它血小板聚集剂。进一步研究了其消除人vWf-血小板相互作用的能力，发现ATA通过干扰vWf而不是血小板来阻断vWf/GPIb通路，是预防血栓形成早期的潜在工具。人胶原蛋白结合结构域：发现了一株能抑制vWf与人Ⅲ型纤维胶原结合的单克隆抗体（NMC/vW 3），其表位位于vWf亚基上。也就是说，该表位位于氨基酸残基911和1365之间。血管性血友病患者血浆血管性血友病因子分析本研究建立了几种vWf功能检测方法，并对正常人和几种类型的血管性血友病患者血浆vWf进行了vWf蛋白分析。建立了使用微量血浆的vWf-人胶原蛋白结合试验。发现Ⅱ型vWd胶原结合异常.建立了微量样品分析血浆vWf亚基的方法。现在，我们将研究几种类型的vWd中的等离子vWf。