权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Study on the specificity and thrombogenicproperty of anficardiolipin antibodies in serafrom patients with the antiphospholipid syndrome.

抗磷脂综合征患者血清中抗心磷脂抗体的特异性和血栓形成特性研究。

基本信息

批准号：
08670505
负责人：
ICHIKAWA Kenji
金额：
$ 1.34万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

1) Nine clones of IgM monoclonal beta2-glycoprotein I (beta2-GPI) dependent anticardiolipin antibodies (aCL) were established from peripheral blood lymphocytes. These monoclonal aCL recognized beta2-GPI structurally altered by binding to anionic phospholipids.2) Reactivity of monoclonal aCL derived from patients with the APS and from APS prone mice with domain deleted mutants of beta2-GPI.Most of monoclonal aCL were considered to recognize epitope (s) related to 4th domain of beta2-GPI.3) Three dimensional structure of human beta2-GPI was analogized from the tertial structure of the Factor H,using GENETYX-SV/R and QUANTA/CHARMm, which were developed to analyze tertial structure of peptides and proteins.4) To identify the epitopes recognized by pathogenic aCL,random peptide libraries were searched with monoclonal antibodies against beta2-GPI.Peptides bound to EY1C8 and EY2C9, human monoclonal aCL,had consensus motifs respectively. Peptides reacted with WBCAL-1, a mouse monoclonal anti-human beta2-GPI antibody, had overlapping consesus sequences.5)To clarify the epitope recognized by monoclonal antibody aganist beta2-GPI,tertial structure model of human beta2-GPI was searched for structures similar to the identified motifs. The putative epitope recognized by EY1C8 and EY2C9 were identified on the 4th domain of beta2-GPI.The epitope was covered by 3rd domain in free from beta2-GPI.The epitope recognized by Cof-18 was near the phospholipid binding site on the 5th domain of beta2-GPI.This may explain the inhibition of the binding of beta2-GPI to anionic phospholipids by Cof-18.

1)从外周血淋巴细胞中建立了9个IgM单克隆β 2-糖蛋白I（β 2-GPI）依赖性抗心磷脂抗体（aCL）克隆。这些单克隆aCL通过与阴离子磷脂结合而识别结构改变的β 2-GPI。2）来自APS患者和来自APS易感小鼠的单克隆aCL与β 2-GPI的结构域缺失突变体的反应性。大多数单克隆aCL被认为识别与β 2-GPI的第四结构域相关的表位。3）人β 2-GPI的三维结构。利用GENETYX-SV/R和QUANTA/CHARMm软件对H因子的三级结构进行了GPI的模拟。4）为了鉴定致病性aCL识别的表位，用抗β_2-GPI单克隆抗体对随机肽库进行搜索，发现与人单克隆aCL EY_1C_8和EY_2C_9结合的肽段分别具有相同的基序。5）为了明确抗β 2-GPI单克隆抗体识别的表位，在人β 2-GPI的三维结构模型中寻找与所识别的基序相似的结构。EY 1C 8和EY 2C 9识别的抗原表位位于β 2-GPI的第4结构域，在游离状态下被第3结构域所覆盖，Cof-18识别的抗原表位位于β 2-GPI第5结构域的磷脂结合位点附近，这可能是Cof-18抑制β 2-GPI与阴离子磷脂结合的原因。