Morgenthaler N G, Pampel I, Aust G, Seissler J, Scherbaum W A
Department of Internal Medicine III, University of Leipzig, Germany.
Horm Metab Res. 1998 Mar;30(3):162-8. doi: 10.1055/s-2007-978858.
The importance of bioassays measuring stimulating and blocking autoantibodies to the TSH-receptor (TSH-R) by their effect on cAMP production in CHO cells transfected with the recombinant TSH-R is increasingly recognized. The standard technique for this bioassay is cumbersome, as it involves purification of serum IgG with polyethylene glycol (PEG) and resuspension in hypotonic buffer. We have therefore established a simpler approach for the detection of stimulating and blocking autoantibodies using JP09 CHO cells and unfractionated human serum. The cAMP concentration was measured by a highly sensitive commercial radioimmuno assay. Thyroid stimulating autoantibodies (TSAb) were present in 107 out of 126 patients with Graves' disease (85%) and in 4 out of 40 patients with Hashimoto's thyroiditis (10%). Specificity was confirmed by the fact that only 1 patient with insulin dependent diabetes mellitus (IDDM) out of 64 patients with different non-thyroid autoimmune disorders (46 with IDDM, 10 with stiff man syndrome and 8 with rheumatoid arthritis) and 2 out of 100 healthy controls (2%) were positive in this assay. In the subgroup of hyperthyroid Graves' disease patients 76 out of 83 (92%) had TSAb and the same number had TSH binding inhibiting immunoglobulin (TBII), as assessed by the commercial TRAK assay. Although both antibody types showed only a weak correlation (r = 0.30), a combination of TSAb and TBII detected 98% of all Graves' patients and 99% of the hyperthyroid subgroup. Thyroid blocking autoantibodies (TBAb) were measured in 4 out of 24 TSAb negative patients with Graves' disease (17%), who were hypothyroid and positive for TBII. A comparison of our bioassay with the standard bioassay using PEG precipitation showed a good correlation (r = 0.76,p < 0.001), demonstrating the feasibility of the simplified assay for the routine detection of TSAb and TBAb in Graves' disease.
通过对转染重组促甲状腺激素受体（TSH-R）的CHO细胞中cAMP产生的影响来测定刺激和阻断促甲状腺激素受体自身抗体（TSH-R）的生物测定法的重要性日益得到认可。这种生物测定的标准技术很繁琐，因为它涉及用聚乙二醇（PEG）纯化血清IgG并在低渗缓冲液中重悬。因此，我们建立了一种更简单的方法，使用JP09 CHO细胞和未分级的人血清来检测刺激和阻断自身抗体。通过高灵敏度的商业放射免疫测定法测量cAMP浓度。126例格雷夫斯病患者中有107例（85%）存在甲状腺刺激自身抗体（TSAb），40例桥本甲状腺炎患者中有4例（10%）存在。在64例患有不同非甲状腺自身免疫性疾病的患者（46例胰岛素依赖型糖尿病（IDDM）、10例僵人综合征和8例类风湿性关节炎）中，只有1例IDDM患者，以及100例健康对照中有2例（2%）在该测定中呈阳性，证实了其特异性。在甲状腺功能亢进的格雷夫斯病患者亚组中，83例中有76例（92%）有TSAb，通过商业TRAK测定法评估，相同数量的患者有促甲状腺激素结合抑制免疫球蛋白（TBII）。虽然这两种抗体类型仅显示出弱相关性（r = 0.30），但TSAb和TBII的组合检测出了98%的所有格雷夫斯病患者和99%的甲状腺功能亢进亚组患者。在24例TSAb阴性的格雷夫斯病患者中有4例（17%）检测到甲状腺阻断自身抗体（TBAb），这些患者甲状腺功能减退且TBII呈阳性。将我们的生物测定法与使用PEG沉淀的标准生物测定法进行比较，显示出良好的相关性（r = 0.76，p < 0.001），证明了简化测定法用于格雷夫斯病中TSAb和TBAb常规检测的可行性。
