Anticitrullinated protein antibodies facilitate migration of synovial tissue-derived fibroblasts
Anticitrullinated protein antibodies facilitate migration of synovial tissue-derived fibroblasts
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DOI：
10.1136/annrheumdis-2018-214967
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发表时间：
2019-12-01
影响因子：
27.4
通讯作者：
Catrina, Anca Irinel
中科院分区：
文献类型：
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作者：
Sun, Meng;Rethi, Bence;Catrina, Anca Irinel
目的类风湿关节炎(RA)特异性抗瓜氨酸蛋白/肽抗体(AC-PAS)在关节炎症发生前可能与骨丢失和关节痛有关。方法从RA患者滑膜中分离出成纤维细胞样滑膜细胞(FLS)。用从RA患者外周血中提纯的多克隆抗CCP-2抗体(抗CCP-2抗体)或来自单个滑液B细胞的单克隆AC PAS刺激FLS培养。我们通过测量细胞黏附和迁移率以及细胞因子的产生来分析AC PAS是如何调节FLS的。用免疫荧光法分析精氨酸脱亚胺酶(PAD)的蛋白表达和蛋白瓜氨酸化，并用免疫印迹法研究信号转导。结果饥饿应激或趋化因子IL-8对FLS的刺激是增强细胞对AC PAS敏感性的关键。这些挑战导致PAD表达和蛋白瓜氨酸化增加，并通过一种涉及磷酸肌醇3-激酶激活的机制，由AC PA介导诱导FLS迁移。抑制PAD酶或与可溶性瓜氨酸蛋白或多肽竞争可完全阻断AC PA诱导的FLS迁移。不同的单抗AC PA在成纤维细胞和破骨细胞中均可触发不同的细胞效应，提示单个AC PA克隆在疾病发病机制中具有独特的作用。结论在一定的AC PA谱系存在的情况下，一过性滑膜损伤可能导致AC PA介导的FLS迁移增加。
Objectives Rheumatoid arthritis (RA)-specific anti-citrullinated protein/peptide antibodies (AC PAs) might contribute to bone loss and arthralgia before the onset of joint inflammation. We aimed to dissect additional mechanisms by which AC PAs might contribute to development of joint pathology.Methods Fibroblast-like synoviocytes (FLS) were isolated from the synovial membrane of patients with RA. The FLS cultures were stimulated with polyclonal AC PAs (anti-CCP-2 antibodies) purified from the peripheral blood of patients with RA or with monoclonal AC PAs derived from single synovial fluid B cells. We analysed how AC PAs modulate FLS by measuring cell adhesion and mobility as well as cytokine production. Expression of protein arginine deiminase (PAD) enzymes and protein citrullination were analysed by immunofluorescence, and signal transduction was studied using immunoblotting.Results Challenge of FLS by starvation-induced stress or by exposure to the chemokine interleukin-8 was essential to sensitise the cells to AC PAs. These challenges led to an increased PAD expression and protein citrullination and an AC PA-mediated induction of FLS migration through a mechanism involving phosphoinositide 3-kinase activation. Inhibition of the PAD enzymes or competition with soluble citrullinated proteins or peptides completely abolished the AC PA-induced FLS migration. Different monoclonal AC PAs triggered distinct cellular effects in either fibroblasts or osteoclasts, suggesting unique roles for individual AC PA clones in disease pathogenesis.Conclusion We propose that transient synovial insults in the presence of a certain pre-existing AC PA repertoire might result in an AC PA-mediated increase of FLS migration.