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Development of clinical assay method for human chymase

人食糜酶临床检测方法的建立

基本信息

批准号：
10670690
负责人：
URATA Llidenori
金额：
$ 2.11万
依托单位：
Fukuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

The original objectives of the project was 1) to develop Westernblot analysis and biochemical assay for plasma human chymase, 2) to determine clinical significance of plasma chymase measurement, 3) to develop RIA method for human chymase to handle massive clinical samples, 4) to compare plasma chymase levels in various cardiovascular diseases, 5) to determine the association between plasma chymase level and various cardiovascular diseases. Among these objectives, objectives 1 and 2 were completed within the first project year, but objectives 3 has been suspended because the iodinated recombinant h-chymase did not bind with the specific antibody. Therefore, we establish a double sandwich ELISA using monoclonal and polyclonal antibodies for recombinant human chymase. Plasma chymase levels significantly increased in systemic inflammatory diseases (pneumonia autoimmune disease and etc.) Mild level of increase was observed in patients with atherosclerosis and ischemic heart disease. There was a significant positive correlation between plasma chymase level and plasma c-reactive protein or fibrinogen. This fact suggests that plasma chymase level may reflect the levels of tissue inflammation. These findings have been reported in Experimental Biology Meeting (Washington DC, USA) and are under considerations for publication in peer review journal. In order to apply this ELISA to clinical use, improvement of the sensitivity will be necessary.

该项目的最初目标是1）开发血浆食糜酶的Westernblot分析和生化测定，2）确定血浆食糜酶测量的临床意义，3）开发人类食糜酶的RIA方法以处理大量临床样本，4）比较各种心血管疾病中的血浆食糜酶水平，5）确定血浆食糜酶水平与各种心血管疾病之间的关联疾病。其中，目标1和2在第一个项目年内完成，但目标3因碘化重组h-糜酶未与特异性抗体结合而被暂停。因此，我们使用重组人食糜酶的单克隆和多克隆抗体建立了双夹心 ELISA。全身炎症性疾病（肺炎、自身免疫性疾病等）血浆糜烂酶水平显着升高，动脉粥样硬化、缺血性心脏病患者血浆糜烂酶水平轻度升高。血浆食糜酶水平与血浆C反应蛋白或纤维蛋白原呈显着正相关。这一事实表明血浆食糜酶水平可能反映组织炎症水平。这些发现已在实验生物学会议（美国华盛顿特区）上报告，并正在考虑在同行评审期刊上发表。为了将该ELISA应用于临床，需要提高灵敏度。