Overexpression of Toll-like Receptors 3 and 4 in Synovial Tissue From Patients With Early Rheumatoid Arthritis Toll-like Receptor Expression in Early and Longstanding Arthritis
Overexpression of Toll-like Receptors 3 and 4 in Synovial Tissue From Patients With Early Rheumatoid Arthritis Toll-like Receptor Expression in Early and Longstanding Arthritis
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DOI：
10.1002/art.24140
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发表时间：
2008-12-01
影响因子：
--
通讯作者：
Kyburz, Diego
中科院分区：
文献类型：
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作者：
Ospelt, Caroline;Brentano, Fabia;Kyburz, Diego
客观的。分析滑膜和皮肤成纤维细胞中 Toll 样受体 (TLR) 1-10 的表达、调控和生物学相关性，并确定早期类风湿性关节炎 (RA)、长期 RA 和骨关节炎 (OA) 患者滑膜组织中 TLR 2、3 和 4 的表达水平。方法。通过实时聚合酶链式反应 (PCR) 分析 RA 滑膜成纤维细胞 (RASF)、OASF 和皮肤成纤维细胞中 TLR 1-10 的表达。用肿瘤坏死因子 α、白细胞介素 1 β (IL-1 β)、细菌脂肽、聚 (I-C)、脂多糖和鞭毛蛋白刺激成纤维细胞。通过酶联免疫吸附测定测定 IL-6 的产生，并通过实时 PCR 诱导 TLR 2-5、基质金属蛋白酶 (MMP) 3 和 13 信使 RNA。通过免疫组织化学分析滑膜组织中TLRs 2-4的表达。结果。滑膜成纤维细胞表达 TLR 1-6，但不表达 TLR 7-10。在表达的TLR中，TLR-3和TLR-4在滑膜成纤维细胞中最丰富，并且用TLR-3配体聚(I-C)刺激滑膜成纤维细胞导致IL-6、MMP-3和MMP-13最显着的增加。相比之下，皮肤成纤维细胞在受到任何测试刺激后并没有上调 MMP-3 或 MMP-13。在早期 RA 患者的滑膜组织中，TLR-3 和 TLR-4 高表达，并且与长期 RA 患者的水平相当。与 OA 中的表达水平相比，这些表达水平有所升高。结论。我们发现 TLR 高表达，特别是 TLR；如图3和图4所示，在RA的早期阶段，滑膜成纤维细胞体外对TLR配体的反应性表明，导致持续炎症和关节破坏的TLR信号通路在疾病过程的早期被激活。
Objective. To analyze the expression, regulation, and biologic relevance of Toll-like receptors (TLRs) 1-10 in synovial and skin fibroblasts and to determine the expression levels of TLRs 2, 3, and 4 in synovial tissues from patients with early rheumatoid arthritis (RA), longstanding RA, and osteoarthritis (OA).Methods. Expression of TLRs 1-10 in RA synovial fibroblasts (RASFs), OASFs, and skin fibroblasts was analyzed by real-time polymerase chain reaction (PCR). Fibroblasts were stimulated with tumor necrosis factor alpha, interleukin-1 beta (IL-1 beta), bacterial lipopeptide, poly(I-C), lipopolysaccharide, and flagellin. Production of IL-6 was determined by enzyme-linked immunosorbent assay and induction of TLRs 2-5, matrix metalloproteinases (MMPs) 3 and 13 messenger RNA by real-time PCR. Expression of TLRs 2-4 in synovial tissues was analyzed by immunohistochemistry.Results. Synovial fibroblasts expressed TLRs 1-6, but not TLRs 7-10. Among the expressed TLRs, TLR-3 and TLR-4 were the most abundant in synovial fibroblasts, and stimulation of synovial fibroblasts with the TLR-3 ligand poly(I-C) led to the most pronounced increase in IL-6, MMP-3, and MMP-13. In contrast, skin fibroblasts did not up-regulate MMP-3 or MMP-13 after stimulation with any of the tested stimuli. In synovial tissues from patients with early RA, TLR-3 and TLR-4 were highly expressed and were comparable to the levels of patients with longstanding RA. These expression levels were elevated as compared with those in OA.Conclusion. Our findings of high expression of TLRs, particularly TLRs; 3 and 4, at an early stage of RA and the reactivity of synovial fibroblasts in vitro to TLR ligands suggest that TLR signaling pathways resulting in persistent inflammation and joint destruction are activated early in the disease process.