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）功能域的分析以及针对 vWf.A 的单克隆抗体（MAb）的制备和表征。使用DEAE和肝素柱纯化因子VIII结合结构域：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 种不同的针对 p-34 的 MAb 由法国 INSERM 143 的 D.Meyer 与我合作生产。现在，我将描述这些抗体的特征 B。血小板糖蛋白 Ib(GPIb) 结合结构域：对抑制 vWf 与 GPIb 结合的两种不同的 vWf 单克隆抗体（来自我们实验室的 NMC/vW4。来自法国的 322）进行了表征，我发现了与 GPIb 结合的不同功能位点。 vWf 由两种诱导剂引起，例如抗生素 ristocetin 和毒液因子 botrocetin。已知金精三羧酸 (ATA) 可以抑制瑞斯托菌素诱导的血小板聚集，但不能抑制其他血小板聚集剂。进一步研究了其消除人类 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。人胶原结合结构域：发现了一种抑制vWf-人III型纤维胶原结合的单克隆抗体（NMC/vW3），并显示其表位位于vWf亚基中。即，表位位于氨基酸残基911和1365之间。II。冯·维勒布兰德病血浆冯·维勒布兰德因子分析。我研究了几种vWf功能检测方法的建立以及对正常人和几种血管性血友病患者血浆vWf的vWf蛋白分析。建立了使用微量血浆的 vWf-人胶原蛋白结合测定法。发现II型vWd中胶原蛋白结合异常。B．建立了血浆vWf中vWf亚基的微量样品分析方法。现在要检查几种vWd 中的血浆vWf。