Graves' disease is an autoimmune disorder, which is characterized by stimulatory antibodies targeting the human thyrotropin receptor (TSHR), resulting in hyperthyroidism and multiple organ damage. We systematically investigated monomeric and dimeric fusion proteins of the A subunit of TSHR for efficacy to bind to the monoclonal patient antibody M22, to interact with Graves' patient serum samples, and to impact on anti-TSHR antibody titers, hyperthyroidism, tachycardia and other in vivo read-outs in a long-term mouse model of Graves' disease induced by immunization with a recombinant adenovirus encoding TSHR A. Binding assays and functional measurements of TSHR-dependent cAMP formation showed binding of monomeric TSHR-His and dimeric TSHR-Fc to the anti-TSHR antibody M22 at low-effective concentrations (EC50 of 5.7 nmol/L and 8.6 nmol/L) and inhibition of the effects of this antibody at high efficiencies (IC50 values of 16-20 nmol/L). Both proteins also block the effects of polyclonal anti-TSHR antibodies occurring in Graves' patient sera with somewhat lower average efficiencies (mean IC50 values of 29 nmol/L and 68 nmol/L). However, in vivo characterization of epicutaneous patch administrations of TSHR-Fc at doses of 0.3 and 0.6 mg/kg body weight in a murine Graves' disease model did not result in any improvement of disease parameters. In conclusion, high affinity binding of TSHR-Fc to pathological anti-TSHR antibodies was not matched by efficacy to improve Graves' disease parameter in a long-term mouse model.

中文翻译：

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格雷夫斯病中的促甲状腺激素受体（TSHR）融合蛋白。

格雷夫斯病是一种自身免疫性疾病，其特征是针对人促甲状腺激素受体（TSHR）的刺激性抗体，导致甲状腺功能亢进和多器官损伤。我们系统地研究了TSHR A亚基的单体和二聚体融合蛋白与单克隆患者抗体M22结合，与Graves患者的血清样品相互作用以及对抗TSHR抗体滴度，甲状腺功能亢进，心动过速等的影响。用编码TSHR A的重组腺病毒免疫诱导的Graves病长期小鼠模型中的体内读数。结合测定和TSHR依赖性cAMP形成的功能测量显示，单体TSHR-His和二聚体TSHR-Fc结合至低浓度的抗TSHR抗体M22（EC50为5.7 nmol / L和8。6 nmol / L）和高效率抑制该抗体的作用（IC50值为16-20 nmol / L）。两种蛋白均以较低的平均效率（平均IC50值为29 nmol / L和68 nmol / L）阻断了出现在Graves患者血清中的多克隆抗TSHR抗体的作用。然而，在鼠格雷夫斯病模型中以0.3和0.6mg / kg体重的剂量对TSHR-Fc的表皮贴剂的体内表征并未导致疾病参数的任何改善。总之，在长期小鼠模型中，TSHR-Fc与病理性抗TSHR抗体的高亲和力结合无法改善Graves病参数的功效。两种蛋白均以较低的平均效率（平均IC50值为29 nmol / L和68 nmol / L）阻断了出现在Graves患者血清中的多克隆抗TSHR抗体的作用。然而，在鼠格雷夫斯病模型中以0.3和0.6mg / kg体重的剂量对TSHR-Fc的表皮贴剂的体内表征并未导致疾病参数的任何改善。总之，在长期小鼠模型中，TSHR-Fc与病理性抗TSHR抗体的高亲和力结合无法改善Graves病参数的功效。两种蛋白均以较低的平均效率（平均IC50值为29 nmol / L和68 nmol / L）阻断了出现在Graves患者血清中的多克隆抗TSHR抗体的作用。然而，在鼠格雷夫斯病模型中以0.3和0.6mg / kg体重的剂量对TSHR-Fc的表皮贴剂的体内表征并未导致疾病参数的任何改善。总之，在长期小鼠模型中，TSHR-Fc与病理性抗TSHR抗体的高亲和力结合无法改善Graves病参数的功效。在鼠格雷夫斯病模型中6 mg / kg体重未导致疾病参数的任何改善。总之，在长期小鼠模型中，TSHR-Fc与病理性抗TSHR抗体的高亲和力结合无法改善Graves病参数的功效。在鼠格雷夫斯病模型中6 mg / kg体重未导致疾病参数的任何改善。总之，在长期小鼠模型中，TSHR-Fc与病理性抗TSHR抗体的高亲和力结合无法改善Graves病参数的功效。