A novel monoclonal antibody recognizing extracellular space of the atherosclerotic lesions in WHHL rabbits.

一种新型单克隆抗体，可识别 WHHL 兔动脉粥样硬化病变的细胞外空间。

基本信息

批准号：
03454495
负责人：
TAKANO Tatsuya
金额：
$ 4.1万
依托单位：
Teikyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

项目摘要

In order to study the mechanisms of cholesteryl ester accumulation in the extracellular space of the atherosclerotic aorta, we prepared novel monoclonal antibodies against atherosclerotic lesions using a delipidated crude homogenate of atherosclerotic aorta from Watanabe-heritable hyperlipidemic (WHHL) rabbits as a complex mixture of immunogens. By screening the antibodies histochemically, we were able to obtain for the first time monoclonal antibodies which specifically recognized the extracellular matrix with lipid-laden deposits (EMRla/212D) in atherosclerotic lesions. The purified antigenic material is a glycoprotein with a molecular mass of 66 kDa, and the epitope of the antigenic material contains sialic acid as a major element.We have isolated from rabbit liver cDNA library clones coding for the 66-kDa glycoprotein (GP66). The clone spans the sequence coding for the entire GP66 (456 amino acids) and 19 amino acids of a signal peptide. GP66 was deduced to be rabbit vitronectin from its nucleotide sequence, containing an Arg-Gly-Asp cell attachment sequence and showing 76% amino acid sequence homology with human vitronectin. Furthermore, EMRla/212D recognized rabbit vitronectin purified by heparin affinity chromatography. RNA blot hybridization detected one transcript of the same size in normal and WHHL rabbit liver. The levels of plasma GP66 and liver GP66 mRNA were not altered, whereas 9-fold greater accumulation of GP66 was observed in the thoracic aorta of WHHL rabbit. These results suggest that GP66 is rabbit vitronectin, which selectively accumulates in the atherosclerotic aorta with the development of atherosclerosis.

为了研究动脉粥样硬化主动脉的细胞外空间中胆固醇酯的积累机制，我们使用deallated heraberable herabyable高光（WHHL）的混合物混合了一种动脉粥样硬化主动脉的差异均质均质的粗型主动脉，制备了针对动脉粥样硬化病变的新型单克隆抗体。通过在组织化学上筛选抗体，我们能够首次获得单克隆抗体，这些抗体特异性地识别了动脉粥样硬化病变中含有脂质沉积物（EMRLA/212D）的细胞外基质。纯化的抗原材料是一种糖蛋白，分子质量为66 kDa，抗原材料的表位含有唾液酸为主要元素。我们已经从编码66 kDa糖蛋白的兔肝cDNA图书馆克隆中分离出来。克隆跨越编码整个GP66（456个氨基酸）和19个信号肽的氨基酸的序列。将GP66从其核苷酸序列中推导为兔玻璃蛋白，其中含有Arg-Gly-Asp细胞附着序列，并显示了与人玻璃体蛋白的76％氨基酸序列同源性。此外，Emrla/212d识别通过肝素亲和力色谱纯化的兔玻璃蛋白。 RNA印迹杂交检测到一个正常和WHHL兔肝脏中相同大小的转录本。血浆GP66和肝GP66 mRNA的水平没有改变，而在WHHL兔的胸主动脉中观察到了9倍GP66的积累。这些结果表明GP66是兔玻璃纤维，随着动脉粥样硬化的发展，它们在动脉粥样硬化主动脉中有选择地积累。