Uno C, Nishikawa M
Second Division of Internal Medicine, Kansai Medical University, Osaka, Japan.
Nihon Naibunpi Gakkai Zasshi. 1988 Mar 20;64(3):193-205. doi: 10.1507/endocrine1927.64.3_193.
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. Porcine thyroid cells after digestion were cultured for 15-18 hours with TSH of 1-10,000 microU/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 0.5 ml of 30% PEG solution to 0.5 ml serum, and was resuspended with 0.6 ml of Hanks' medium without NaCl, containing 1.5% bovine serum albumin, 20mM Hepes and 0.5 mM 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 40 microU/ml. When the PEG precipitations from 10 normal subjects were incubated with the thyroid cells of 4 X 10(5) 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 incubated with normal IgG was judged as positive TSAb activity. The minimum detectable quantities were regarded as about 5 microU/ml TSH equivalent. 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 less than 0.01) in patients with untreated Graves' disease. In these patients, TSAb activities showed a significant positive correlation with values for 99mTc thyroid uptake, determined 30 min after the injection. However, they did not show any significant correlation with serum T4 or T3 concentrations. Similarly, TBII showed significant correlations with goiter size and 99mTc thyroid uptake. To conclude, the present assay for TSAb is sensitive and reproducible.(ABSTRACT TRUNCATED AT 400 WORDS)
采用灵敏的检测方法，利用培养的猪甲状腺细胞以及用聚乙二醇（PEG）从血清中沉淀的方法，测定格雷夫斯病患者血清中促甲状腺素抗体（TSAb）的活性，并将其活性与采用商业检测试剂盒测定的促甲状腺素结合抑制免疫球蛋白（TBII）的活性进行比较。消化后的猪甲状腺细胞与1 - 10,000微单位/毫升的促甲状腺激素（TSH）或正常受试者、格雷夫斯病患者及桥本甲状腺炎患者血清的沉淀物一起培养15 - 18小时，然后用商业放射免疫分析试剂盒（山佐）测定培养基中的环磷酸腺苷（cAMP）水平。通过向0.5毫升血清中加入0.5毫升30%的PEG溶液获得沉淀物，并用0.6毫升不含氯化钠的汉克斯培养基重悬，该培养基含有1.5%的牛血清白蛋白、20毫摩尔/升的羟乙基哌嗪乙磺酸（Hepes）和0.5毫摩尔/升的3 - 异丁基 - 1 - 甲基黄嘌呤。当原始血清中TSH含量超过40微单位/毫升时，沉淀物中含有约85%的原始血清免疫球蛋白、63%的白蛋白以及大量TSH。当将10名正常受试者的PEG沉淀物与4×10⁵个细胞的甲状腺细胞一起孵育时，培养基中环磷酸腺苷的释放量在83%至124%之间，以平均值计算为100%。因此，与正常免疫球蛋白（IgG）孵育的培养基中释放量超过130%的环磷酸腺苷释放被判定为TSAb阳性活性。最低可检测量被视为约5微单位/毫升促甲状腺素当量。在52例未经治疗的甲状腺功能亢进格雷夫斯病患者中，分别检测到48例（92%）的TSAb活性和50例（96%）的TBII活性，在20例通过抗甲状腺药物治疗维持临床甲状腺功能正常状态的格雷夫斯病患者中，16例（80%）检测到TSAb或TBII活性。其中10例（50%）患者的TBII呈阳性。尽管未经治疗的格雷夫斯病患者中TSAb和TBII活性之间存在显著正相关（r = 0.53，p < 0.01），但一些患者在这两种活性上表现出明显差异。在这些患者中，TSAb活性与注射后30分钟测定的99mTc甲状腺摄取值呈显著正相关。然而，它们与血清T4或T3浓度无显著相关性。同样，TBII与甲状腺肿大小和99mTc甲状腺摄取呈显著相关。总之，目前的TSAb检测方法灵敏且可重复。（摘要截短于400字）
