Shanghai Clinical Center for Endocrine and Metabolic Diseases, Department of Endocrinology and Metabolism, Ruijin Hospital, Affiliated to Shanghai Jiao-Tong University School of Medicine, Shanghai, China.
PLoS One. 2013 May 22;8(5):e64277. doi: 10.1371/journal.pone.0064277. Print 2013.
Graves' disease (GD) is a common autoimmune disease involving the thyroid gland. The altered balance of pro- and anti-inflammatory cytokines plays an important role in the pathogenesis of GD. Chemokine (C-C motif) ligand 20 (CCL20) is important for interleukin-17 (IL-17) signal activation and a potent chemoattractant for Th17 cells. Meanwhile, Osteopontin (OPN), a broadly expressed pleiotropic cytokine, has been implicated in GD through inducing Th1-involved response to enhance the production of proinflammatory cytokines and chemokines, but little is known about the role of OPN in regulating CCL20 and IL-17 signaling.
This study sought to explore the possibility of CCL20 level as a biomarker for GD, as well as investigate the role of OPN in regulating CCL20 production.
Fifty untreated GD patients, fifteen euthyroid GD patients, twelve TRAb-negative GD patients and thirty-five healthy control donors were recruited. OPN, CCL20 and other clinical GD diagnosis parameters were measured. CD4+T cells were isolated from peripheral blood mononuclear cells (PBMCs) using antibody-coated magnetic beads. Enzyme-linked immune-sorbent assay and quantitative polymerase chain reaction were used to determine CCL20 expression level.
We found that the plasma CCL20 level was enhanced in GD patients and decreased in euthyroid and TRAb-negative GD patients. In addition, CCL20 level correlated with GD clinical diagnostic parameters and plasma OPN level. Moreover, we demonstrated that recombinant OPN and plasma from untreated GD patients increased the expression of CCL20 in CD4+T cells, which could be blocked by OPN antibody. Furthermore, we found that the effect of OPN on CCL20 expression was mediated by β3 integrin receptor, IL-17, NF-κB and MAPK pathways.
These results demonstrated that CCL20 might serve as a biomarker for GD and suggested the possible role of OPN in induction of CCL20 expression.
格雷夫斯病（GD）是一种常见的自身免疫性甲状腺疾病。促炎和抗炎细胞因子的平衡改变在 GD 的发病机制中起重要作用。趋化因子（C-C 基序）配体 20（CCL20）对于白细胞介素 17（IL-17）信号的激活很重要，并且是 Th17 细胞的有效趋化因子。同时，骨桥蛋白（OPN）作为一种广泛表达的多功能细胞因子，通过诱导 Th1 参与反应来增强促炎细胞因子和趋化因子的产生，从而与 GD 有关，但对于 OPN 调节 CCL20 和 IL-17 信号的作用知之甚少。
本研究旨在探讨 CCL20 水平作为 GD 生物标志物的可能性，并研究 OPN 在调节 CCL20 产生中的作用。
招募了 50 名未经治疗的 GD 患者、15 名甲状腺功能正常的 GD 患者、12 名 TRAb 阴性的 GD 患者和 35 名健康对照供体。测量了 OPN、CCL20 和其他 GD 诊断参数。使用抗体包被的磁珠从外周血单核细胞（PBMCs）中分离 CD4+T 细胞。酶联免疫吸附试验和实时定量聚合酶链反应用于确定 CCL20 的表达水平。
我们发现 GD 患者的血浆 CCL20 水平升高，甲状腺功能正常和 TRAb 阴性的 GD 患者的 CCL20 水平降低。此外，CCL20 水平与 GD 临床诊断参数和血浆 OPN 水平相关。此外，我们证明重组 OPN 和未经治疗的 GD 患者的血浆增加了 CD4+T 细胞中 CCL20 的表达，而 OPN 抗体可阻断该作用。此外，我们发现 OPN 对 CCL20 表达的作用是通过β3 整合素受体、IL-17、NF-κB 和 MAPK 途径介导的。
这些结果表明 CCL20 可能作为 GD 的生物标志物，并提示 OPN 在诱导 CCL20 表达中的可能作用。
