STUDY ON THE FUNCTIONAL DOMAINS OF VON WILLEBRAND FACTOR IN PATIENTS WITH VON WILLEBRAND DISEASE

血管性血友病患者血管性血友病因子功能域的研究

• 批准号: 06670820
• 负责人: TAKAHASHI Yukihiro
• 金额: $ 1.15万
• 依托单位: NARA MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670820/
• 关键词: von Willebrand factor; von Willebrand disease; factor VIII; ron Willebrand disease

The functional domains of von Willebrand favtor (vWf)(1) The amino-terminal fragment of vWf (III-T4), which includes factor VIII-binding domain, was purified from the trypsin-digestions of vWf. Five monoclonal antibodies (MAbs) agaist to vWf were produced and were determined the epitopes on vWf. Epitopes of four MAbs (464,418,511,522) were located in III-T4, one (516) did not recognized on III-T4. Four MAbs (464,418,522,516) inhibited the binding of factor VIII to vWf. MAb (418) recognized on the conformational structure of vWf. On the other hand, MAb (522) detected on the linear structure of vWf. Now, we analyze further closed epitopes of these MAbs.(2) We found a patient with plasma vWf which was not reacted with MAb (522). We are going to study further investigation of this plasma vWF.2. The analysis of plasma vWf in patients with various types of von willebrand disease (vWd)(1) We established the method of the analysis for reduced vWf using a minute amount of plasma without purification.(2) We analyzed reduced plasma vWf in patients with various types of vWd and discoverd two patients with abnormal proteolytic reduced fragment in type I vWd.(3) These patients with abnormal reduced fragment in type I vWd have a point mutaion, Arg611 to His.(4) We have now researched the family members of these patients with abnormal reduced plasma vWf and was assayd the functional activities of plasma vWf.

vWf的功能结构域（1）从vWf的胰蛋白酶-双酶切产物中纯化出vWf的氨基端片段（III-T4），该片段包括因子VIII结合结构域。制备了5株抗vWf的单克隆抗体，并测定了vWf的抗原表位。其中4株（464、418、511、522）位于III-T4区，1株（516）未被识别。4种单克隆抗体（464、418、522、516）抑制因子VIII与vWf的结合。MAb（418）在vWf的构象结构上被识别。另一方面，在vWf的线性结构上检测到MAb（522）。现在，我们进一步分析这些单克隆抗体的封闭表位。(2)我们发现1例患者血浆vWf与MAb无反应（522）。我们将进一步研究这种血浆vWF。2。不同类型血管性血友病（vWd）患者血浆vWf的分析（1）建立了不经纯化的微量血浆vWf分析方法。(2)我们分析了不同类型vWd患者的血浆vWf降低，并对两例I型vWd蛋白水解还原片段异常的患者进行了分析。(3)这些I型vWd异常减少片段的患者存在Arg 611 → His点突变。(4)我们对这些血浆vWf异常降低患者的家属进行了研究，并测定了血浆vWf的功能活性。

项目成果

野並京子 他4: "血漿von Willebrand factor の還元フラグメント解析にて異常200kDaバンドを示したtype I vW Willebrand diseaseの家系検索" 血栓止血誌. (投稿予定).

Kyoko Nonami 等人 4：“在血浆冯维勒布兰德因子减少片段分析中显示异常 200kDa 条带的 I 型 vW 血友病家族搜索”《血栓形成与止血杂志》（待提交）。

Kyoko NONAMI,Yukihiro TAKAHASHI,Toshiya NISIKUBO Naoyuki MORII,Sajji KINOSHITA,Sachiko NISHIDA Takayuki NISHIYAMA,Yoshihiro FUJIMURA,Akira YOSHIOKA: "Reduced plasma von Willebrand factor fragments in patients with type 1, type 2A and type 2B von Willebran

Kyoko NONAMI、Yukihiro TAKAHASHI、Toshiya NISIKUBO Naoyuki MORII、Sajji KINOSHITA、Sachiko NISHIDA Takayuki NISHIYAMA、Yoshihiro FUJIMURA、Akira YOSHIOKA：“1 型、2A 型和 2B 型血管性血友病患者血浆血管性血友病因子片段减少

野並京子 他8: "von Willebrand病名種病型における血漿von Willebrand因子の還元フラグメントの検討" 血栓止血誌. (投稿中).

Kyoko Nonami 等人 8：“冯·维勒布兰德疾病类型中血浆冯·维勒布兰德因子减少片段的检查”《血栓形成和止血杂志》（目前正在提交）。

高橋幸博(他1名): 医歯薬出版株式会社(編集 桜川信男,池田康夫), 489 (1994)

高桥幸宏（及其他 1 人）：石药出版株式会社（樱川伸夫、池田康夫编辑），489 (1994)

Kyoko NONAMI,Yukihiro TAKAHASHI,Toshiya NISIKUBO Naoyuki MORII,Akira YOSHIOKA: "Reduced von Willebrand factor fragment in plasma" Jpn.J.Thromb Hemorrh. 6 (4). 279-286 (1995)

Kyoko NONAMI、Yukihiro TAKAHASHI、Toshiya NISIKUBO Naoyuki MORII、Akira YOSHIOKA：“血浆中冯维勒布兰德因子片段减少”Jpn.J.血栓性出血。