Purificaiton, distribution and clinical significance of TSH-receptor in human thyroid.

我们试图制造一种针对人tsh受体的抗体。利用该抗体检测tsh受体的分布及分布变化。首先，我们尝试用高效价tsh受体抗体患者的血清纯化tsh受体抗体。培养含高效价tsh受体抗体的血清和猪甲状腺膜组分。结合IgG用酸性缓冲液释放，释放的IgG用IgG亲和层析纯化。利用该IgG检测了人甲状腺滤泡细胞中tsh受体的定位。顶质膜呈阳性染色。免疫印迹法在mol.wt 130,000中发现了唯一的一条条带。我们还研究了ras p21产物的超微结构定位，该产物参与GTP对腺苷酸环化酶的调控。结果表明，ras p21产物定位于顶端细胞膜。同时用免疫组织化学方法检测了人甲状腺组织中表皮生长因子（EGF）及其受体的表达。这些证据表明，甲状腺滤泡细胞的顶质膜限制了tsh受体。目前对各种甲状腺疾病中tsh受体的分布变化进行了研究。

We have attempted to produce an antibody for human TSH-receptor. Using this antibody, distribution and changes in distribution of TSH-receptor was examined. First, we attempted to purify the TSH-receptor antibody using the serum of patients with high titer of TSH-receptor antibody. The serum containing high titer of TSH-receptor antibody and porcine thyroid membrane fraction were incubated. The binding IgG was released by acidic buffer and released IgG was further purified with IgG affinity chromatography. Using this IgG the localization of TSH-receptor was examined in human thyroid follicular cells. The positive staining was observed at the apical plasma membrane. The only one band was found at Mol. wt 130,000 using the immunoblotting. We also examined the ultrastructural localization of ras p21 product, which participates on GTP regulation of adenylate cyclase. Similar finding that ras p21 product was localized at apical cell membrane ws obtained. Epidermal growth factor (EGF) and EGF-receptor were also examined immunohistochemically in human thyroid. These combined evidences strongly suggested that TSH-receptor is restricted at apical plasma membrane of thyroid follicular cells. The changes in distribution of TSH-receptro in various thyroid diseases have been studied at present.