Expression of Toll-like Receptor 3 and Toll-like Receptor 7 in Muscle Is Characteristic of Inflammatory Myopathy and Is Differentially Regulated by Th1 and Th17 Cytokines
Expression of Toll-like Receptor 3 and Toll-like Receptor 7 in Muscle Is Characteristic of Inflammatory Myopathy and Is Differentially Regulated by Th1 and Th17 Cytokines
复制标题
DOI：
10.1002/art.27465
复制
发表时间：
2010-07-01
影响因子：
--
通讯作者：
Miossec, P.
中科院分区：
文献类型：
--
作者：
Tournadre, A.;Lenief, V.;Miossec, P.
Objective.目的检测多发性肌炎（PM）和皮肌炎（DM）患者肌肉组织中TLR-3和TLR-7的表达，探讨TLR-3在培养的肌肉细胞中的功能和调节。使用免疫组织化学分析TLR-3、TLR-7、HLA I类和CD 56（未成熟成肌细胞前体的标志物）的表达。在存在或不存在中和性抗TLR-3抗体的情况下，在成肌细胞和分化的肌管中用TLR-3激动剂聚（I-C）、坏死的成肌细胞以及Th 1和Th 17细胞因子评估TLR-3调节和信号传导。通过逆转录-聚合酶链反应定量TLR-3信使RNA（mRNA）的水平。采用酶联免疫吸附法测定白细胞介素-6（IL-6）、CCL 20和IL-8的水平。TLR-3和TLR-7在PM/DM组织中表达，但在非炎性肌肉组织中不表达，并且主要在炎性浸润中检测到，尽管少数肌肉细胞也呈阳性。这些TLR-3和TLR-7阳性纤维表达高水平的CD 56和HLA I类抗原。对于IL-6和CCL 20的产生，观察到聚（I-C）和IL-17之间的协同作用。类似地，用坏死成肌细胞刺激增加IL-6产生，并且用坏死成肌细胞与IL-17组合刺激进一步增加IL-6的诱导。TLR-3阻断可降低坏死成肌细胞和IL-17对IL-6产生的诱导作用。干扰素γ（IFN γ）刺激可增加TLR-3 mRNA的表达，但IL-17可下调IFN γ的诱导作用。我们的研究结果表明，TLR-3和TLR-7在炎症性肌病组织中表达，特别是在未成熟的成肌细胞前体中。坏死的肌肉细胞激活细胞因子的产生，部分通过TLR-3途径，具有Th 1和Th 17细胞因子的差异调节作用。
Objective. To assess the expression of Toll-like receptor 3 (TLR-3) and TLR-7 in muscle tissue from patients with polymyositis (PM) and dermatomyositis (DM) and to investigate the function and regulation of TLR-3 in cultured muscle cells.Methods. The expression of TLR-3, TLR-7, HLA class I, and CD56, a marker of immature myoblast precursors, was analyzed using immunohistochemistry. TLR-3 regulation and signaling were assessed in myoblasts and in differentiated myotubes with the TLR-3 agonist poly(I-C), necrotic myoblasts, and Th1 and Th17 cytokines, in the presence or absence of neutralizing anti-TLR-3 antibody. Levels of TLR-3 messenger RNA (mRNA) were quantified by reverse transcription-polymerase chain reaction. Levels of interleukin-6 (IL-6), CCL20, and IL-8 were determined by enzyme-linked immunosorbent assay.Results. TLR-3 and TLR-7 were expressed in PM/DM tissues, but not in noninflammatory muscle tissues, and were primarily detected in inflammatory infiltrates, although a few muscle cells were also positive. These TLR-3-and TLR-7-positive fibers expressed high levels of CD56 and HLA class I antigens. A synergy between poly(I-C) and IL-17 was observed for the production of IL-6 and CCL20. Similarly, stimulation with necrotic myoblasts increased IL-6 production, and stimulation with necrotic myoblasts in combination with IL-17 further increased the induction of IL-6. TLR-3 blockade decreased the inducing effect of necrotic myoblasts and IL-17 on IL-6 production. Stimulation with interferon-gamma (IFN gamma) increased TLR-3 mRNA levels, but IL-17 down-regulated the inducing effect of IFN gamma.Conclusion. Our findings indicate that TLR-3 and TLR-7 are expressed in inflammatory myopathic tissues, particularly in immature myoblast precursors. Necrotic muscle cells activate cytokine production, in part, through the TLR-3 pathway, with a differential regulatory effect of Th1 and Th17 cytokines.