Graves' Autoantibodies Exhibit Different Stimulating Activities in Cultures of Thyrocytes and Orbital Fibroblasts Not Reflected by Clinical Assays.
Graves' Autoantibodies Exhibit Different Stimulating Activities in Cultures of Thyrocytes and Orbital Fibroblasts Not Reflected by Clinical Assays.
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格雷夫斯自身抗体在甲状腺细胞和眼眶成纤维细胞培养物中表现出不同的刺激活性，但临床检测并未反映出这一点。
DOI：
10.1089/thy.2021.0326
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Gershengorn,MarvinC
中科院分区：
文献类型：
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作者：
Krieger,ChristineC;Kahaly,GeorgeJ;Azam,Asma;Klubo-Gwiezdzinska,Joanna;Neumann,Susanne;Gershengorn,MarvinC
背景：Graves甲亢（GH）和相关的Graves眼窝病（GO）的发病机制似乎与刺激性自身抗体（促甲状腺素受体[TSHR]刺激抗体[TSAbs]）有关，该抗体结合并激活甲状腺细胞和眼眶成纤维细胞上的TSHR。一般来说，临床检测循环TSHR抗体与生长激素和氧化石墨烯的状态相关。然而，大多数TSHR抗体的临床测量使用竞争结合试验，不区分TSHR抗体和结合但不激活TSHR的抗体。此外，在非甲状腺细胞或眼眶成纤维细胞的工程细胞中，TSAbs的临床试验仅测量了一种信号通路的刺激，即环腺苷单磷酸(cAMP)-蛋白激酶A （PKA）通路。我们确定在工程细胞中通过cAMP-PKA读数测量TSAbs是否准确地揭示了这些抗体对甲状腺细胞和眼眶成纤维细胞的刺激效果。方法：在体外原代培养中，我们检测了TSAb对正常人甲状腺细胞和氧化石墨烯患者眼眶成纤维细胞的刺激作用。在甲状腺细胞中，我们测量了甲状腺球蛋白（TG）的分泌和眼眶成纤维细胞透明质酸（HA）的分泌。我们还测量了工程tshrr表达细胞中cAMP产生的刺激，实验方法类似于临床试验。此外，我们确定单纯GH患者的tsab对甲状腺细胞和眼眶成纤维细胞的刺激与理解GO患者伴有GH的GO患者的tsab是否存在差异。结果：我们发现TSAb刺激工程细胞cAMP生成与甲状腺细胞分泌TG和眼眶成纤维细胞分泌HA呈正相关。然而，GH患者的tsb比GO患者的tsb更有效地刺激甲状腺细胞，而GO患者的tsb比GH患者的tsb更有效地激活眼眶成纤维细胞。结论：tabs测量cAMP-PKA通路激活的临床刺激与甲状腺细胞和眼眶成纤维细胞的刺激相关；然而，他们没有区分GH和GO患者的tsab。在体外，TSAbs在激活TSHRs方面表现出选择性，因为GO患者的TSAbs在刺激眼眶成纤维细胞方面更有效，而GH患者的TSAbs在刺激甲状腺细胞方面更有效。
Background:The pathogenesis of Graves' hyperthyroidism (GH) and associated Graves' orbitopathy (GO) appears to involve stimulatory autoantibodies (thyrotropin receptor [TSHR]-stimulating antibodies [TSAbs]) that bind to and activate TSHRs on thyrocytes and orbital fibroblasts. In general, measurement of circulating TSHR antibodies by clinical assays correlates with the status of GH and GO. However, most clinical measurements of TSHR antibodies use competitive binding assays that do not distinguish between TSAbs and antibodies that bind to but do not activate TSHRs. Moreover, clinical assays for TSAbs measure stimulation of only one signaling pathway, the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway, in engineered cells that are not thyrocytes or orbital fibroblasts. We determined whether measuring TSAbs by a cAMP-PKA readout in engineered cells accurately reveals the efficacies of stimulation by these antibodies on thyrocytes and orbital fibroblasts.Methods:We measured TSAb stimulation of normal human thyrocytes and orbital fibroblasts from patients with GO in primary culturesin vitro. In thyrocytes, we measured secretion of thyroglobulin (TG) and in orbital fibroblasts secretion of hyaluronan (hyaluronic acid [HA]). We also measured stimulation of cAMP production in engineered TSHR-expressing cells in an assay similar to clinical assays. Furthermore, we determined whether there were differences in stimulation of thyrocytes and orbital fibroblasts by TSAbs from patients with GH alone versus from patients with GO understanding that patients with GO have accompanying GH.Results:We found a positive correlation between TSAb stimulation of cAMP production in engineered cells and TG secretion by thyrocytes as well as HA secretion by orbital fibroblasts. However, TSAbs from GH patients stimulated thyrocytes more effectively than TSAbs from GO patients, whereas TSAbs from GO patients were more effective in activating orbital fibroblasts than TSAbs from GH patients.Conclusions:Clinical assays of stimulation by TSAbs measuring activation of the cAMP-PKA pathway do correlate with stimulation of thyrocytes and orbital fibroblasts; however, they do not distinguish between TSAbs from GH and GO patients.In vitro, TSAbs exhibit selectivity in activating TSHRs since TSAbs from GO patients were more effective in stimulating orbital fibroblasts and TSAbs from GH patients were more effective in stimulating thyrocytes.