Heufelder A E, Wenzel B E, Gorman C A, Bahn R S
Department of Medicine, Mayo Clinic, Rochester, Minnesota 55905.
J Clin Endocrinol Metab. 1991 Oct;73(4):739-45. doi: 10.1210/jcem-73-4-739.
We hypothesize that fibroblasts obtained from the retroocular space and the pretibial skin, sites affected by the peripheral manifestations of Graves' disease, share unique characteristics that may in part explain the site specificity of Graves' ophthalmopathy (GO) and pretibial myxedema (PTM). Heat shock proteins (HSPs), synthesized by cells undergoing stress, function to maintain cellular homeostasis and are probably involved in the intracellular processing and cell surface presentation of antigens. We investigated possible differences in the expression of 70-kDa HSPs between cultured fibroblasts obtained from patients with severe GO and normal individuals. In addition, we compared HSP expression in fibroblasts derived from tissues involved in the extrathyroidal manifestation of Graves' disease (GO and PTM) with that in fibroblasts from uninvolved tissues. HSPs were detected by both immunoblotting and indirect immunofluorescence, using monoclonal antibodies that are directed against HSP72, HSP72/73 (termed HSP70), and HSP90. HSP expression at baseline and after treatment with various cytokines and heat stress was examined. At baseline, HSP72 reactivity was exclusively detected in retroocular and pretibial fibroblasts from patients with severe GO and PTM, but was not observed in abdominal fibroblasts from these patients and was not detectable in fibroblasts from any anatomical site of normal individuals. The abundance of HSP70 expression at baseline and after treatment with certain cytokines was significantly greater in retroocular and pretibial fibroblasts from patients with GO than in normal individuals. In addition, characteristic changes in the cellular localization of HSPs before and after exposure to heat stress and cytokines were observed; cell surface expression of HSP70 was detected at baseline in fibroblasts from patients, but not in normal fibroblasts. These data provide the first evidence that HSPs are differentially expressed by fibroblasts derived from tissues affected by the extrathyroidal manifestations of GD. These proteins may have a role in localized immune processes, leading to the development of GO and PTM.
我们推测，从眼球后间隙和胫前皮肤获取的成纤维细胞（这些部位受格雷夫斯病外周表现的影响）具有独特特征，这可能部分解释了格雷夫斯眼病（GO）和胫前黏液性水肿（PTM）的部位特异性。热休克蛋白（HSPs）由经历应激的细胞合成，其功能是维持细胞内稳态，可能参与抗原的细胞内加工和细胞表面呈递。我们研究了严重GO患者培养的成纤维细胞与正常个体培养的成纤维细胞之间70-kDa热休克蛋白表达的可能差异。此外，我们比较了格雷夫斯病甲状腺外表现（GO和PTM）所涉及组织来源的成纤维细胞与未受累组织来源的成纤维细胞中的热休克蛋白表达。使用针对HSP72、HSP72/73（称为HSP70）和HSP90的单克隆抗体，通过免疫印迹和间接免疫荧光检测热休克蛋白。检测了基线时以及用各种细胞因子和热应激处理后的热休克蛋白表达。在基线时，仅在严重GO和PTM患者的眼球后和胫前成纤维细胞中检测到HSP72反应性，但在这些患者的腹部成纤维细胞中未观察到，在正常个体任何解剖部位的成纤维细胞中也未检测到。GO患者的眼球后和胫前成纤维细胞在基线时以及用某些细胞因子处理后的HSP70表达丰度显著高于正常个体。此外，观察到热应激和细胞因子暴露前后热休克蛋白细胞定位的特征性变化；患者来源的成纤维细胞在基线时检测到HSP70的细胞表面表达，而正常成纤维细胞中未检测到。这些数据首次证明，格雷夫斯病甲状腺外表现所影响组织来源的成纤维细胞中热休克蛋白存在差异表达。这些蛋白质可能在局部免疫过程中起作用，导致GO和PTM的发生。
