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.

冯维勒布兰德因子 (vWf) 的功能域(1) 从 vWf 的胰蛋白酶消化中纯化出 vWf (III-T4) 的氨基末端片段，其中包括因子 VIII 结合域。制备了五种针对 vWf 的单克隆抗体 (MAb)，并测定了 vWf 上的表位。四种 MAb (464,418,511,522) 的表位位于 III-T4，其中一种 (516) 在 III-T4 上未识别。四种 MAb (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，MAb (522)检测到vWf的线性结构。现在，我们进一步分析这些MAb的封闭表位。(2)我们发现一名血浆vWf患者不与MAb发生反应(522)。我们将进一步研究该血浆 vWF.2。各种类型血管性血友病（vWd）患者血浆vWf的分析（1）我们建立了使用微量未经纯化的血浆进行还原vWf的分析方法。（2）我们分析了vWd患者的还原血浆vWf。并发现2例I型vWd蛋白水解还原片段异常的患者。(3)这些I型vWd蛋白水解还原片段异常的患者存在点突变，Arg611变为(4)我们现在对这些血浆vWf异常降低的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。