DEK in the synovium of patients with juvenile idiopathic arthritis: characterization of DEK antibodies and posttranslational modification of the DEK autoantigen.
DEK in the synovium of patients with juvenile idiopathic arthritis: characterization of DEK antibodies and posttranslational modification of the DEK autoantigen.
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DOI：
10.1002/art.30138
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发表时间：
2011-02
影响因子：
--
通讯作者：
Markovitz, David M.
中科院分区：
文献类型：
--
作者：
Mor-Vaknin, Nirit;Kappes, Ferdinand;Dick, Amalie E.;Legendre, Maureen;Damoc, Catalina;Teitz-Tennenbaum, Seagal;Kwok, Roland;Ferrando-May, Elisa;Adams, Barbara S.;Markovitz, David M.
DEK是幼年特发性关节炎（JIA）患儿的一种核磷蛋白和自身抗原。在与异常免疫激活相关的广泛疾病中也发现了针对DEK的自身抗体。我们先前证明DEK由巨噬细胞分泌，由凋亡T细胞释放，并吸引白细胞。由于我们在JIA患者的滑液中发现了DEK，我们现在研究DEK蛋白和/或自身抗体如何参与JIA的发病机制。DEK自身抗体，免疫复合物，和滑膜巨噬细胞从JIA患者的滑液纯化。DEK自身抗体和免疫复合物通过亲和柱层析纯化，并通过二维凝胶电泳、免疫印迹和ELISA进行分析。通过连续离心和磁珠纯化上清液和外泌体中的DEK，并通过Nano-LC-MS/MS鉴定DEK的翻译后修饰。在JIA患者的滑液中发现DEK自身抗体和蛋白。DEK由滑膜巨噬细胞以游离形式和通过外泌体分泌。DEK自身抗体（IgG 2）可以激活补体级联反应，主要识别DEK蛋白的C-末端部分，并对乙酰化DEK表现出更高的亲和力。与这些观察结果一致，DEK经历了前所未有数量的赖氨酸残基上的乙酰化，如Nano-LC-MS/MS所示。这些结果表明，DEK可以通过DEK和DEK自身抗体之间的高亲和力相互作用产生免疫复合物，直接促进JIA中的关节炎症，该过程通过DEK在发炎关节中的乙酰化而增强。
DEK is a nuclear phosphoprotein and autoantigen in a subset of children with juvenile idiopathic arthritis (JIA). Autoantibodies to DEK are also found in a broad spectrum of disorders associated with abnormal immune activation. We previously demonstrated that DEK is secreted by macrophages, released by apoptotic T cells, and attracts leukocytes. As we identified DEK in synovial fluids in JIA patients, we now investigate how DEK protein and/or autoantibodies may contribute to the pathogenesis of JIA. DEK autoantibodies, immune complexes, and synovial macrophages were purified from synovial fluids of JIA patients. DEK autoantibodies and immune complexes were purified by affinity column chromatography and analyzed by 2-D gel electrophoresis, immunoblotting and ELISA. DEK in supernates and exosomes was purified by serial centrifugation and magnetic beads, and DEK’s posttranslational modifications were identified by Nano-LC-MS/MS. DEK autoantibodies and protein are found in synovial fluids from JIA patients. DEK is secreted by synovial macrophages in a free form and via exosomes. DEK autoantibodies (IgG2) may activate the complement cascade, primarily recognize the C-terminal portion of DEK protein and exhibit higher affinity for acetylated DEK. Consistent with these observations, DEK undergoes acetylation on an unprecedented number of lysine residues as demonstrated by Nano-LC-MS/MS. These results indicate that DEK can contribute directly to joint inflammation in JIA by generating immune complexes through high affinity interaction between DEK and DEK autoantibodies, a process enhanced by acetylation of DEK in the inflamed joint.
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影响因子：
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