Chen Peng, Yan Hai, Tian Yaping, Xun Yiping, Shi Lili, Bao Ran, Zhang Huai, Chen Guangyu, Yang Chunhe, Sun Shutao, Wang Yajie, Liu Li, Zhou Yabin, Zhang Chunyan, Wang Xiaoxu, Wen Yongqiang, Bian Yongzhong, Du Hongwu
School of Chemistry and Biotechnology Engineering, University of Science and Technology Beijing, Beijing 100083, China.
Department of Clinical Biochemistry, Chinese PLA General Hospital, Beijing 100853, China.
Sci Rep. 2015 Feb 2;5:8162. doi: 10.1038/srep08162.
Cell membrane proteins are believed to play a critical role in the pathogenesis of autoimmune diseases. However, few membrane autoantigens have been linked with Behçet's disease. Here, a cell-chip was performed to identify autoantibody target cells, and the suspected autoantigens were detected using immunoblotting. The amino acid sequences of the detected proteins were determined using LC-MALDI-TOF/TOF. Putative proteins were recombinantly expressed and purified, and a corresponding ELISA was developed and clinically validated using real clinical samples. It was found that a 36-kDa membrane protein--annexin A2--was detected in approximately one-third of the patients' blood circulation. The immunohistochemistry results showed that annexin A2 was highly expressed in vascular endothelial cells. Moreover, vascular involvement was significantly higher in the anti-annexin A2 antibody-positive group versus the anti-annexin A2 antibody-negative group among all the clinical samples analyzed, indicating that annexin A2 is a novel endothelial cell membrane antigen involved in Behçet's disease.
细胞膜蛋白被认为在自身免疫性疾病的发病机制中起关键作用。然而，很少有膜自身抗原与白塞病相关联。在此，进行了细胞芯片实验以鉴定自身抗体靶细胞，并使用免疫印迹法检测可疑的自身抗原。使用液相色谱 - 基质辅助激光解吸电离飞行时间/串联飞行时间质谱（LC-MALDI-TOF/TOF）确定所检测蛋白质的氨基酸序列。对推定的蛋白质进行重组表达和纯化，并开发了相应的酶联免疫吸附测定（ELISA），并使用实际临床样本进行临床验证。结果发现，在大约三分之一的患者血液循环中检测到一种36 kDa的膜蛋白——膜联蛋白A2。免疫组织化学结果显示膜联蛋白A2在血管内皮细胞中高表达。此外，在所有分析的临床样本中，抗膜联蛋白A2抗体阳性组的血管受累情况明显高于抗膜联蛋白A2抗体阴性组，表明膜联蛋白A2是一种参与白塞病的新型内皮细胞膜抗原。
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