Okuda J, Akamizu T, Sugawa H, Matsuda F, Hua L, Mori T
Department of Laboratory Medicine Faculty of Medicine, Kyoto University, Japan.
J Clin Endocrinol Metab. 1994 Dec;79(6):1600-4. doi: 10.1210/jcem.79.6.7989462.
Anti-TSH receptor antibodies (TSH-R Ab), which have been detected in the serum of some patients with primary myxedema, are themselves considered to induce hypothyroidism. These are termed blocking-type TSH-R Ab (TSH-R BAb), because they inhibit adenylate cyclase stimulation by TSH on thyrocytes or nonthyroidal cells transfected with TSH-R complementary DNA. We prepared monoclonal TSH-R BAb and characterized them. Peripheral lymphocytes from three patients with primary hypothyroidism and potent TSH-R BAb were transformed by Epstein-Barr virus, and the culture supernatants were screened by TSH binding inhibitor immunoglobulin (TBII) assay. Twenty positive and 7 negative lymphocyte clones were obtained; their monoclonality was confirmed by Southern blot analysis, using an immunoglobulin (Ig) JH probe. These monoclonal antibodies were then tested for TSH-R BAb activity. TSH-R BAb activity ranged from 24.1-58.5% (normal range, < 24%) in all 20 TBII-positive clones and in 2 of 7 TBII-negative clones. An enzyme-linked immunosorbent assay showed that the Ig isotypes of these clones with TBII and/or TSH-R BAb activity were IgG in 8 and IgM in 14. Another enzyme-linked immunosorbent assay and Southern blot analysis of the light chains revealed that 13 clones had kappa-chains, whereas the light chains could not be determined in the other 9 clones. To summarize, 1) we obtained 22 clones that produced monoclonal TSH-R BAb, including 8 IgG-type clones. 2) The clones exhibited dominant usage of the kappa-chain. 3) Although all TBII clones had TSH-R BAb activity, their TBII and TSH-R BAb activities were not significantly correlated, and two TSH-R BAb clones did not show TBII activity.
抗促甲状腺激素受体抗体（TSH-R Ab）已在一些原发性黏液性水肿患者的血清中被检测到，其本身被认为可诱发甲状腺功能减退。这些抗体被称为阻断型TSH-R Ab（TSH-R BAb），因为它们可抑制促甲状腺激素对甲状腺细胞或转染了TSH-R互补DNA的非甲状腺细胞中腺苷酸环化酶的刺激作用。我们制备了单克隆TSH-R BAb并对其进行了特性分析。来自三名原发性甲状腺功能减退患者且含有高效TSH-R BAb的外周淋巴细胞经EB病毒转化，培养上清液通过促甲状腺激素结合抑制免疫球蛋白（TBII）检测进行筛选。获得了20个阳性和7个阴性淋巴细胞克隆；使用免疫球蛋白（Ig）JH探针通过Southern印迹分析证实了它们的单克隆性。然后对这些单克隆抗体进行TSH-R BAb活性检测。在所有20个TBII阳性克隆以及7个TBII阴性克隆中的2个克隆中，TSH-R BAb活性范围为24.1% - 58.5%（正常范围 < 24%）。酶联免疫吸附测定显示，这些具有TBII和/或TSH-R BAb活性的克隆的Ig同种型中，8个为IgG，14个为IgM。另一种酶联免疫吸附测定以及轻链的Southern印迹分析显示，13个克隆具有κ链，而其他9个克隆无法确定轻链。总之，1）我们获得了22个产生单克隆TSH-R BAb的克隆，其中包括8个IgG型克隆。2）这些克隆表现出κ链的优势使用。3）尽管所有TBII克隆都具有TSH-R BAb活性，但它们的TBII和TSH-R BAb活性并无显著相关性，并且有两个TSH-R BAb克隆未显示出TBII活性。
