Rapoport B, Filetti S, Takai N, Seto P, Halverson G
Metabolism. 1982 Nov;31(11):1159-67. doi: 10.1016/0026-0495(82)90168-8.
Studies were conducted on the cultured human thyroid cell bioassay for thyroid stimulating immunoglobulin (TSI) and thyrotropin (TSH). In confirmation of the data of Kasagi, et al.,15 incubation of human thyroid cells in Hank's balanced salt solution deficient in NaCl increased the sensitivity of the c-AMP response to bovine TSH by approximately one order of magnitude. Half-maximal stimulation was attained at approximately 0.1 mU bTSH/ml. The effect of NaCl in the medium was greater with stimulation by TSI greater than hTSH greater than bTSH. In contract to incubations in NaCl(+)medium, with NaCl(-) medium most (70%-80%) of the c-AMP produced was released into the medium; this proportion remaining relatively constant over a wide range of bTSH and hTSH concentrations. At TSI concentrations higher than 3mg/ml efflux of c-AMP into the medium was significantly diminished. Stimulation by cholera toxin and prostaglandin E of thyroid cell c-AMP generation was not enhanced in NaCl(-) medium, in contrast to stimulation by TSH and TSI. The presence of 10(-4)m cycloheximide in NaCl(+) medium enhanced the c-AMP response to physiological concentrations of TSH. As with NaCl(-) medium, cycloheximide increased the sensitivity but not the maximum response of the c-AMP response to TSH. However no additivity was observed with NaCl(-) medium and cycloheximide. Human thyroid cells obtained from patients with Graves' disease are relatively insensitive to TSI stimulation. In NaCl(-) medium, however, the sensitivity of these cells to TSI stimulation is sufficient to enable them to be utilized in the TSI assay. The present state of the TSI assay is discussed.
针对促甲状腺素免疫球蛋白（TSI）和促甲状腺激素（TSH）开展了人甲状腺细胞培养生物测定研究。为证实笠木等人的数据，在不含NaCl的汉克平衡盐溶液中培养人甲状腺细胞，可使细胞对牛TSH的c - AMP反应敏感性提高约一个数量级。在约0.1 mU bTSH/ml时达到最大刺激的一半。培养基中NaCl的影响在TSI刺激时比人TSH刺激时比牛TSH刺激时更大。与在含NaCl培养基中孵育相比，在不含NaCl培养基中，产生的c - AMP大部分（70% - 80%）释放到培养基中；在较宽的bTSH和人TSH浓度范围内，该比例保持相对恒定。在TSI浓度高于3mg/ml时，c - AMP向培养基中的外流量显著减少。与TSH和TSI刺激不同，在不含NaCl培养基中，霍乱毒素和前列腺素E对甲状腺细胞c - AMP生成的刺激未增强。在含NaCl培养基中存在10(-4)m放线菌酮可增强对生理浓度TSH的c - AMP反应。与不含NaCl培养基一样，放线菌酮提高了c - AMP对TSH反应的敏感性，但未提高最大反应。然而，在不含NaCl培养基和放线菌酮之间未观察到相加作用。从格雷夫斯病患者获得的人甲状腺细胞对TSI刺激相对不敏感。然而，在不含NaCl培养基中，这些细胞对TSI刺激的敏感性足以使其用于TSI测定。文中讨论了TSI测定的现状。
