Monoclonal Thyroid Stimulating Antibodies

促甲状腺激素受体(TSHR)是Graves病的主要抗原，也是甲状腺刺激的靶点 在这类患者的血清中发现的TSHR抗体(TSHR-Ab)。TSHR-Ab的特点是作用时间长 最初被称为长效甲状腺刺激剂(LATS)。在此应用程序中，我们寻求继续我们的 成功克隆和鉴定了抗TSH受体的单抗，这是第一个 具有刺激活性的单抗。特别是，单抗MS-1已显示出强大的TSHR 当用稳定表达hTSHR的CHO细胞测试时，刺激活性明显下降到2 ng/mlIg G。 这次竞争性更新的具体目标是：目标1：确定更多的刺激甲状腺的单抗 抗体。我们将首先继续在我们成功的基础上开发一系列扩大的单抗 与具有促甲状腺活性的TSH受体有关。目的2：定义TSHR-AB表位。我们将描述 TSH受体抗体的线性和非线性抗原表位确定主要免疫原性 TSH受体上的‘区域(MIR)。目标3：定义抗体介导的TSHR翻译后调节 修改。我们将在CHO细胞中共表达TSHRcpp TSHRmyc结构，并检测TSHRcpp在CHO细胞中是否占据 TSHR-Abs的受体分子将调节FRET指数，就像我们已经看到的TSH配体一样。此外，我们还有 最近开发了一种独特的测定TSHR切割的方法，并将检测TSHR-Abs对TSHR-Abs的影响 受体切割，目的是深入了解这种切割在甲状腺自身免疫中的功能作用。 目的：检测TSHR-Abs的生物学功能及信号转导。我们将研究 TSHR-Abs的生长刺激/抑制能力和激活/抑制功能的特异性 分化的甲状腺细胞功能。 与日本血吸虫TSHR-Abs特性相似的TSH抗体的分离与鉴定 Graves病患者的血清将对其作用机制提供重大的新见解，并提供新的 控制甲状腺功能障碍的方法。

The thyrotropin (TSH) receptor (TSHR) is the primary antigen of Graves' disease and the target for thyroid stimulating 'TSHR antibodies (TSHR-Ab)found in the serum of such patients. TSHR-Ab are characterized by their prolonged action and were originally called Long Acting Thyroid Stimulators (LATS). In this application, we seek to continue our successful cloning and characterization of monoclonal antibodies to the TSH receptor which have resulted in the first monoclonal antibody with stimulating activity. In particular, monoclonal antibody MS-1 has shown potent TSHR stimulating activity when tested with CHO cells stably expressing hTSHR with activity evident down to 2ng/ml ofIgG. The specific aims of this competitive renewal are: Aim 1: Identify additional monoclonal thyroid-stimulating antibodies. We will first continue to build on our success by developing an expanded panel of monoclonal antibodies to the TSH receptor with thyroid stimulating activity. Aim 2: Define TSHR-AB epitopes. We will characterize the linear and non-linear antigenic epitopes for the derived TSH receptor antibodies to define the major immunogenic 'regions (MIRs) on the TSH receptor. Aim 3: Defining antibody-mediated regulation of TSHR post-translational modifications. We will co-express TSHRcpp TSHRmycconstructs in CHO cells and examine whether occupation of the receptor molecule by TSHR-Abs will modulate the FRET index as we have seen for TSH ligand. In addition, we have recently developed a unique assay for measuring TSHR cleavage and will examine the influence of TSHR-Abs on receptor cleavage with the aim of gaining insight into the functional role of such cleavage in thyroid autoimmunity. Aim 4: Examine the biologic function and signal transduction induced by TSHR-Abs. We will examine the specificity of TSHR-Abs with regard to their growth stimulating/inhibiting potential and their activarion/inacrivation of differentiated thyroid cell function. The isolation and characterization of antibodies to the TSH with similar characteristics to the TSHR-Abs found in the serum of Graves' disease patients will provide major new insight into their mechanisms of action and offer novel approaches to the control of thyroid dysfunction.