Citrullinated fibrinogen detected as a soluble citrullinated autoantigen in rheumatoid arthritis synovial fluids
Citrullinated fibrinogen detected as a soluble citrullinated autoantigen in rheumatoid arthritis synovial fluids
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DOI：
10.1136/ard.2005.044743
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发表时间：
2006-08-01
影响因子：
27.4
通讯作者：
Yamamoto, K.
中科院分区：
文献类型：
--
作者：
Takizawa, Y.;Suzuki, A.;Yamamoto, K.
背景：抗瓜氨酸化蛋白抗体（ACPA）在类风湿性关节炎（RA）患者血清中特异性且频繁检测到。瓜氨酸纤维蛋白或纤维蛋白原是这类抗体的候选自身抗原。目的：探讨类风湿关节炎（RA）患者和对照组血浆或滑液中瓜氨酸纤维蛋白原（cFBG）的存在，并确定cFBG水平及其与RA血清标志物的关系。方法：采用纤维蛋白原A α链衍生序列R16Cit和R252Cit（瓜氨酸在16位和252位）分别免疫小鼠产生的单克隆抗体cF16.1和cF252.1，建立夹心酶联免疫吸附法（ELISA）检测cFBG，并采用免疫沉淀-western blotting进一步检测cFBG的存在。结果：使用cF16.1和抗修饰瓜氨酸（AMC）抗体的夹心ELISA检测cFBG，在11/15 RA滑膜液（rasf）中检测到阳性信号，而在骨关节炎滑膜液或RA血浆中检测不到阳性信号。免疫沉淀-western blotting证实rasf中存在cFBG。此外，大多数RA血清对R16Cit有强烈反应。配对血清的C反应蛋白或抗环瓜氨酸肽抗体水平与RASF cFBG水平没有关系。结论：cFBG在rasf中可作为可溶性瓜氨酸化自身抗原检测到，因此可能是RA患者ACPA的真正候选抗原。
Background: Anti-citrullinated protein antibodies ( ACPA) are specifically and frequently detected in sera of patients with rheumatoid arthritis ( RA). Citrullinated fibrin or fibrinogen is a candidate autoantigen of such antibodies.Objective: To investigate the presence of citrullinated fibrinogen ( cFBG) in the plasma or synovial fluid of patients with RA and control patients, and to determine cFBG levels and their relationship with serum markers for RA if it is present.Methods: A sandwich enzyme linked immunosorbent assay ( ELISA) to measure cFBG was established using monoclonal antibodies cF16.1 and cF252.1, generated by immunising mice with R16Cit and R252Cit, the fibrinogen A alpha chain derived sequences with citrulline at position 16 and 252, respectively, and the presence of cFBG was further investigated with immunoprecipitation-western blotting. Results: Positive signals were detected in 11/15 RA synovial fluids ( RASFs), but not in osteoarthritis synovial fluids or RA plasma with sandwich ELISA for cFBG using cF16.1 and an anti-modified citrulline ( AMC) antibody. The presence of cFBG in RASFs was confirmed by immunoprecipitation-western blotting. Furthermore, most RA sera strongly reacted against R16Cit. No relationship was seen between RASF cFBG levels and C reactive protein or anti-cyclic citrullinated peptide antibody levels of the paired sera.Conclusion: cFBG is detected as a soluble citrullinated autoantigen in RASFs and may therefore be a genuine candidate antigen for ACPA in patients with RA.