Synovial fluid is a site of citrullination of autoantigens in inflammatory arthritis
Synovial fluid is a site of citrullination of autoantigens in inflammatory arthritis
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DOI：
10.1002/art.23618
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发表时间：
2008-08-01
影响因子：
--
通讯作者：
Venables, Patrick J.
中科院分区：
文献类型：
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作者：
Kinloch, Andrew;Lundberg, Karin;Venables, Patrick J.
目标。目的：检测类风湿关节炎(RA)患者滑液中产生瓜氨酸抗原(包括候选自身抗原瓜氨酸α-烯醇化酶)的部位。20例类风湿关节炎(RA)、20例脊柱炎(SpA)和20例骨关节炎(OA)患者的滑液。样品用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法分离，考马斯亮蓝染色和免疫印迹检测瓜氨酸蛋白、α-烯醇化酶和脱亚胺酶PAD-2和PAD-4。用双向电泳法分离RA滑液样品中的蛋白质，并用免疫印迹和质谱仪鉴定每个蛋白质。采用酶联免疫吸附试验检测抗瓜氨酸α-烯醇化酶多肽1(CEP-1)和环瓜氨酸多肽2(CVP-2)抗体。在RA和SpA患者的滑液中检测到瓜氨酸多肽，而在OA样本中未检测到瓜氨酸多肽。所有样本均检测到α-烯醇化酶，类风湿性关节炎患者L平均水平为6.4 ng/亩，温泉患者样本平均水平为4.3 ng/亩L；
Objective. To examine synovial fluid as a site for generating citrullinated antigens, including the candidate autoantigen citrullinated alpha-enolase, in rheumatoid arthritis (RA).Methods. Synovial fluid was obtained from 20 patients with RA, 20 patients with spondylarthritides (SpA), and 20 patients with osteoarthritis (OA). Samples were resolved using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by staining with Coomassie blue and immunoblotting for citrullinated proteins, alpha-enolase, and the deiminating enzymes peptidylarginine deiminase type 2 (PAD-2) and PAD-4. Proteins from an RA synovial fluid sample were separated by 2-dimensional electrophoresis, and each protein was identified by immunoblotting and mass spectrometry. Antibodies to citrullinated alpha-enolase peptide 1 (CEP-1) and cyclic citrullinated peptide 2 were measured by enzyme-linked immunosorbent assay.Results. Citrullinated polypeptides were detected in the synovial fluid from patients with RA and patients with SpA, but not in OA samples. Alpha-enolase was detected in all of the samples, with mean levels of 6.4 ng/mu l in RA samples, 4.3 ng/mu l in SpA samples, and