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Analysis of autoantibodies in the mechanism of the occurrence of multiple autoimmune endocrinopathies

自身抗体在多种自身免疫性内分泌病发生机制中的分析

基本信息

批准号：
06454616
负责人：
KOBAYASHI Isao
金额：
$ 4.03万
依托单位：
Gunma 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：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

We have established an assay method of antipituitary antibodies (APA) as a screening test, using enzyme-linked immunosorbent assay (ELISA) and evaluated on its clinical usefulness. The soluble fractions of rat pituitary cells were used as antigens of ELISA/APA.The assay, similar to that utilized indirect enzyme autoantibodies, was performed. The results by this assay method were calculated as the cut off index (C.I value), using colorimetric reaction and absorvance at 450 nm.Pathiens with autoimmune thyroid diseases (Hashimoto's thyroiditis Graves, disease) showed approximately 20% positive for ELISA/APA.On the other hand, 34% in Hashimoto's thyroiditis and 28% in Graves disease had a positive band by Western blot analysis, which was identical with 22 kDa protein. Furthermore, 39% in patients with insulin-dependent diabetes mellitus (IDDM) was positive for ELISA/APA,in which 64% in such patients was positive by Western blot analysis.We next analyzed 22 kDa protein ontained by Western blot method, which was one of matched antigens to APA.The analysis of amino acid sequence in this protein resulted in P-A-M-L-S-S-L-F-N-A-V-L-R from the N terminal side. Employing homology analysis, this matched antigens was found to be identical with molecular weight of somatotropin.Further studies are needed to determine the nature of other positive bands by Western blot analysis. As reported previously, some patients with pituitary dwarfism and panhypopituitarism showed positive APA.On the basis of these observations, the present study suggests that measurements of APA are a useful indicator of multiple autoimmune endocrinopathies.

我们建立了抗垂体抗体（阿帕）的酶联免疫吸附试验（ELISA）筛选方法，并对其临床应用价值进行了评价。以大鼠垂体细胞可溶性组分为抗原，采用与间接酶自身抗体相似的方法进行ELISA/阿帕检测。计算该测定方法的结果作为临界值指数（C.I值），采用比色反应和450 nm处的扩散。在桥本甲状腺炎和Graves病中，ELISA/阿帕的阳性率分别为20%和34%，而在桥本甲状腺炎和Graves病中分别为28%和28%。与22 kDa蛋白质完全一致。胰岛素依赖型糖尿病（insulin dependent diabetes mellitus，IDDM）患者ELISA/阿帕阳性率为39%，其中Western blot检测阳性率为64%，用Western blot方法检测APA的22 kDa蛋白，该蛋白是阿帕的一种匹配抗原，经氨基酸序列分析，N端为P-A-M-L-S-S-L-F-N-A-V-L-R。经同源性分析，该抗原与生长激素的分子量一致，其余阳性条带的性质有待进一步研究。如前所述，一些垂体性侏儒症和全垂体功能减退症患者的APA呈阳性。在这些观察的基础上，本研究表明，阿帕的测量是一个有用的指标，多种自身免疫性内分泌疾病。