Studies on the mechanism of autoimmune poliglandular endocrine disorders determined ny antipituitary antibodies

我们建立了用免疫印迹法检测抗垂体抗体的方法。并对抗垂体抗体检测的临床意义进行了评价。将50ul的样品(200ug蛋白质)与50ul的Smple缓冲液混合。用半干印迹法将分离的蛋白质转移到PVDF膜上。该膜在1：101稀释的人血清中孵育，经抗垂体抗体识别的垂体抗原鉴定，获得22 kDa的阳性条带。胰岛素依赖型糖尿病患者血清甲状腺抗体阳性率为40.0%(12/30)，而非胰岛素依赖型糖尿病患者无一例阳性。53例Graves病患者中有14例(26.4%)和54例桥本甲状腺炎患者中有12例(22.2%)出现22 kDa阳性条带。在25例垂体性侏儒症患者中有11例抗体阳性(44.0%)，22 kDa蛋白被认为是大鼠垂体胞浆中与生长激素相关的蛋白，因此，我们用Western Blot分析了一种新的抗垂体抗体的检测体系。我们现在正在进行脑垂体细胞表面膜抗体的检测。

We have established an assay method by western Blot analysis of antipituitary antibodies. and evaluated on clinical significance of determination of antipituitary antibodies.Antigens was obtained from rat pituitary cytosolic fractions. 50 ul of sample(200 ug protein) was mixed with 50 ul of smple buffer. Searated proteins were transfered to a PVDF membrane by a semi-dry blotting apparatus. The membrane was incubated in 1 : 101 diluted human serum.We have fund 22 kDa positive band after identification of pituitary antigens recognized by antipituitary antibodies. The prevalence of positive oituitary antibodies by this method was 40.0%(12/30) in patents with insulin-dependent diabetes melltus, while nobody showed positive results in patients withnon-insulin dependent diabetes mellitus. The 22 kDa positive band was found in 14 of 53 patients with Graves' disease(26.4%) and 12 of 54 patients with Hashimoto's thyroditis(22.2%). Eleven of 25 patients with pituitary dwarfism showed positive antibodies(44.0%).The 22 kDa protein was considered to be GH-related protein in rat pituitary cytosolic fractions.Thus, we have demonstrated a new assay system by western Blot analysis of antipituitary antibodies. We are now in progress to determine pituitary cell surface membrane antibodies.