バセドウ病の異常甲状腺刺激物質に関する臨床的検討:第一編 : ブタ甲状腺培養細胞及びPEG抽出IgGを用いた高感度thyroid-stimulating antibody (TSAb) 測定法の基礎的並びに臨床的検討
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The activities of thyroid-stimulating antibody (TSAb) in serum from patients with Graves' disease were measured by a sensitive assay, using cultured porcine thyroid cells and the precipitation from serum with polyethylene glycol (PEG), and the activities were compared with those of thyrotropin binding inhibitor immunoglobulin (TBII), measured by the commercial assay kit.<BR>Porcine thyroid cells after digestion were cultured for 15-18 hours with TSH of 1-10,000μU/ml or the precipitations of sera from normal subjects and patients with Graves' disease or Hashimoto's thyroiditis, and then the cAMP levels in the culture medium were determined by the commercial RIA assay kit (Yamasa). The precipitation was obtained by adding 0.5ml of 30% PEG solution to 0.5ml serum, and was resuspended with 0.6ml of Hanks' medium without NaCl, containing 1.5% bovine serum albumin, 20mM Hepes and 0.5mM 3-isobutyl-1-methylxanthine. The precipitation contained about 85% of immunoglobulin and 63% of albumin of the original amount of the serum, as well as substantial TSH, when the original serum contained TSH more than 40μU/ml.<BR>When the PEG precipitations from 10 normal subjects were incubated with the thyroid cells of 4 × 10<SUP>5</SUP> cells, the cAMP releases into the medium ranged from 83 to 124%, when the mean value was calculated as 100%. Therefore, the cAMP release of more than 130% of the amount released into the culture medium when incubated with normal IgG was judged as positive TSAb activity. The minimum detectable quantities were regarded as about 5μU/ml TSH equivalent.<BR>TSAb and TBII activities were detected in 48 (92%) and 50 (96%) of 52 patients with untreated hyperthyroid Graves' disease, respectively, and either TSAb or TBII activities were detected in 16 (80%) of 20 patients with Graves' disease maintained in a clinically euthyroid state by treatment with antithyroid drugs. TBII was positive in 10 (50%) of these patients. Some patients showed distinct discrepancies in these two activities, although there was a significant positive correlation between TSAb and TBII activities (r=0.53, p<0.01) in patients with untreated Graves' disease. In these patients, TSAb activities showed a significant positive correlation with values for <SUP>99m</SUP>Tc thyroid uptake, determined 30 min after the injection. However, they did not show any significant correlation with serum T<SUB>4</SUB> or T<SUB>3</SUB> concentrations. Similarly, TBII showed significant correlations with goiter size and <SUP>99m</SUP>Tc thyroid uptake.<BR>To conclude, the present assay for TSAb is sensitive and reproducible. Although these two activities are not always equally detected in these patients, the TSAb and/or TBII activities are detected in virtually all patients with untreated hyperthyroid Graves' disease, suggesting that these activities are important for the etiology of Graves' disease.
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