The Expression of toll-like receptors 3 and 7 in rheumatoid arthritis synovium is increased and costimulation of toll-like receptors 3, 4, and 7/8 results in synergistic cytokine production by dendritic cells
The Expression of toll-like receptors 3 and 7 in rheumatoid arthritis synovium is increased and costimulation of toll-like receptors 3, 4, and 7/8 results in synergistic cytokine production by dendritic cells
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DOI：
10.1002/art.21278
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发表时间：
2005-08-01
影响因子：
--
通讯作者：
Radstake, TRDJ
中科院分区：
文献类型：
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作者：
Roelofs, MF;Joosten, LAB;Radstake, TRDJ
Objective.研究类风湿关节炎（RA）患者和健康人滑膜组织中Toll样受体（TLR）3和7的表达，以及TLR-2、TLR-3、TLR-4和TLR-7/8介导的树突状细胞（DCs）成熟和细胞因子产生的差异。采用免疫组织化学方法研究类风湿关节炎（RA）滑液中TLR-3和TLR-7的表达。将来自RA患者和健康对照的单核细胞衍生的DC培养6天，随后通过TLR介导的途径（TLR-2的脂磷壁酸、Pam（3）Cys和成纤维细胞刺激脂肽I，TLR-3的聚[I-C]，TLR-4的脂多糖和额外结构域A，TLR-7/8的R848）刺激48小时。使用流式细胞术测量表型DC成熟。使用Bio-ELISA系统测量肿瘤坏死因子α（TNF α）、白细胞介素-6（IL-6）、IL-10和IL-12的分泌。用表达TLR-2和TLR-4的细胞系检测RA患者血清和关节液中TLR-2和TLR-4配体。TLR-3和TLR-7在RA滑膜中呈高表达。所有TLR配体在RA患者和健康对照的DC之间同样引起表型DC成熟。TLR-2和TLR-4介导的RA患者DC刺激导致炎症介质（TNF α和IL-6）的产生明显高于健康对照组的DC。相反，在TLR-3和TLR-7/8的刺激下，来自RA患者的DC和来自健康对照的DC之间的细胞因子产生水平是相等的。值得注意的是，TLR-3和TLR-7/8刺激均导致向IL-12倾斜的平衡。有趣的是，TLR-4和TLR-3-7/8的组合刺激导致关于炎症介质的产生的显著协同作用。作为一个概念的证明，类风湿关节炎患者的血清和滑液中TLR-4配体增加。TLR参与DC活化和细胞因子产生的调节。我们假设关节中的各种TLR配体同时触发多种TLR，有利于RA耐受性的突破。
Objective. To evaluate the expression of Toll-like receptors (TLRs) 3 and 7 in synovium and to study potential differences in the maturation and cytokine production mediated by TLR-2, TLR-3, TLR-4, and TLR-7/8 by dendritic cells (DCs) from rheumatoid arthritis (RA) patients and DCs from healthy controls.Methods. Synovial expression of TLR-3 and TLR-7 in RA was studied using immunohistochemistry. Monocyte-derived DCs from RA patients and healthy controls were cultured for 6 days and subsequently stimulated for 48 hours via TLR-mediated pathways (lipoteichoic acid, Pam(3)Cys, and fibroblast-stimulating lipopeptide I for TLR-2, poly[I-C] for TLR-3, lipopolysaccharide and extra domain A for TLR-4, and R848 for TLR-7/8). Phenotypic DC maturation was measured using flow cytometry. The secretion of tumor necrosis factor alpha (TNF alpha), interleukin-6 (IL-6), IL-10, and IL-12 was measured using the Bio-Plex system. Cell lines expressing TLR-2 and TLR-4 were used for the detection of TLR-2 and TLR-4 ligands in serum and synovial fluid from RA patients.Results. TLR-3 and TLR-7 were highly expressed in RA synovium. All TLR ligands elicited phenotypic DC maturation equally between DCs from RA patients and those from healthy controls. TLR-2- and TLR-4-mediated stimulation of DCs from RA patients resulted in markedly higher production of inflammatory mediators (TNF alpha and IL-6) compared with DCs from healthy controls. In contrast, upon stimulation of TLR-3 and TLR-7/8, the level of cytokine production was equal between DCs from RA patients and those from healthy controls. Remarkably, both TLR-3 and TLR-7/8 stimulation resulted in a skewed balance toward IL-12. Intriguingly, the combined stimulation of TLR-4 and TLR-3-7/8 resulted in a marked synergy with respect to the production of inflammatory mediators. As a proof of concept, TLR-4 ligands were increased in the serum and synovial fluid of RA patients.Conclusion. TLRs are involved in the regulation of DC activation and cytokine production. We hypothesize that various TLR ligands in the joint trigger multiple TLRs simultaneously, favoring the breakthrough of tolerance in RA.