Characterization of the optimal stimulatory effects of graves' monoclonal and serum immunoglobulin G on adenosine 3',5'-monophosphate production in fRTL-5 thyroid cells: a potential clinical assay.
Characterization of the optimal stimulatory effects of graves' monoclonal and serum immunoglobulin G on adenosine 3',5'-monophosphate production in fRTL-5 thyroid cells: a potential clinical assay.
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Graves 单克隆和血清免疫球蛋白 G 对 fRTL-5 甲状腺细胞中腺苷 3,5-单磷酸产生的最佳刺激作用的表征：一项潜在的临床测定。
DOI：
10.1210/jcem-57-4-782
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
L. Kohn
中科院分区：
文献类型：
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作者：
P. Vitti;C. Rotella;W. Valente;J. Cohen;S. Aloj;P. Laccetti;F. Ambesi;E. Grollman;A. Pinchera;R. Toccafondi;L. Kohn
来自Graves病甲亢患者血清的免疫球蛋白G(Ig G)制剂和来自Graves‘s淋巴细胞异种杂交瘤融合的单抗Ig G在培养的连续株功能正常的大鼠甲状腺细胞(克隆FRTL-5)中升高cAMP水平。当与5×10(4)个/孔的细胞孵育2小时后，两种类型的免疫球蛋白制剂的最佳刺激作用是相同的，并使用在不生长的、TSH缺乏的培养液中保持7天的细胞。在低浓度下，两种制剂的刺激活性与浓度和相似的Ka值呈线性关系(约4×10~(-8)M)，尽管65%的Graves血清免疫球蛋白制剂具有更强的抑制TSH与膜制剂结合的能力。同一测定中TSH的Ka值约为5×10~(-12)M。使用该细胞分析方法，90%的甲亢Graves‘s免疫球蛋白具有刺激活性，这一值与我们和其他人用人甲状腺细胞和切片系统发现的阳性结果的频率相当。相反，停药1年后甲状腺功能正常的患者(n=21)仅有10%的患者表现出刺激活性，而在非毒性结节性甲状腺肿(n=11)、毒性腺瘤(n=5)和甲状腺癌(n=6)患者中未检测到刺激活性。这些研究表明，优化的大鼠FRTL-5甲状腺细胞系统是检测TSAb的一种临床实用和方便的替代人甲状腺细胞和切片系统。
Immunoglobulin G (IgG) preparations derived from the sera of patients with hyperthyroidism due to Graves' disease (TSAb) as well as a monoclonal IgG derived from heterohybridoma fusions of Graves' lymphocytes augmented cAMP levels in a continuous strain of functioning rat thyroid cells (clone FRTL-5) in culture. Optimal stimulation was the same for both types of IgG preparations when measured after 2 h of incubation with 5 X 10(4) cells/well and using cells maintained in a nongrowth, TSH-deficient medium for 7 days. At low IgG concentrations, the stimulatory activities of both preparations exhibited a linear dependence on concentration and similar Ka values (approximately 4 X 10(-8) M) despite the fact that 65% of the Graves' serum IgG preparations had a significantly better ability to inhibit TSH binding to membrane preparations. The Ka value for TSH in the same assay was about 5 X 10(-12) M. Using this cell assay, 90% of a series of hyperthyroid Graves' IgG preparations exhibited stimulating activity, a value comparable to the frequency of positive results found by ourselves and others using human thyroid cell and slice systems. In contrast, only 10% of patients who were euthyroid 1 yr after antithyroid drug withdrawal (n = 21) exhibited stimulating activity, and no stimulating activity was detected in patients with nontoxic nodular goiter (n = 11), toxic adenoma (n = 5), or thyroid carcinoma (n = 6). The studies suggest that an optimized rat FRTL-5 thyroid cell system is a clinically useful and convenient alternative to human thyroid cell and slice systems for detecting TSAbs.