Chen Peng, Shi Lili, Jiang Yun, Ji Yuting, Yan Hai, Sun Shutao, Xun Yiping, Chen Guangyu, Wang Xiaoxu, Chen Weiyang, Du Hongwu
School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China.
Core Facility, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Biochem Biophys Res Commun. 2015 Jan 24;456(4):866-71. doi: 10.1016/j.bbrc.2014.12.064. Epub 2014 Dec 19.
The aim of this study was to identify candidate pathogenic autoantigens of Behçet's disease (BD) in pathogen-stimulated target cells.
First, three cell lines were used as target cells to screen autoantibody. Second, selected target cells were simulated with pathogens. Third, western blotting was used for detecting the auto-antigens in cell extracts. Next, immunoprecipitation was performed and the amino-acid sequences of target antigens were analyzed by LC-MALDI-TOF/TOF. Then, the potential target antigen was expressed, purified, and immunologically confirmed. And finally, an ELISA kit was developed and clinically validated through the assessments of 456 clinical samples with BD.
One antigen with a molecular weight of approximately 27-kDa was identified as heat shock protein 27 (HSP27). The reactivity of serum IgG against recombinant human HSP27 was detected in 52 of 91 BD patients (57%), 66 of 92 rheumatoid arthritis (RA) patients (72%), 32 of 90 Sjogren syndrome (SS) patients (36%), 22 of 92 systemic lupus erythematosus (SLE) patients (24%) and 0 of 91 healthy controls (HC). The reactivity of BD serum IgG antibodies against HSP27 was significantly higher than SLE (P<0.0001) SS (P<0.0001) and HC (P<0.0001).
This study identified HSP27 as a candidate endothelial cell autoantigen of BD, which is interesting and probably worth further exploration.
本研究旨在鉴定病原体刺激的靶细胞中白塞病（BD）的候选致病性自身抗原。
首先，使用三种细胞系作为靶细胞来筛选自身抗体。其次，用病原体模拟选定的靶细胞。第三，采用蛋白质印迹法检测细胞提取物中的自身抗原。接下来，进行免疫沉淀，并通过液相色谱 - 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）分析靶抗原的氨基酸序列。然后，表达、纯化潜在的靶抗原并进行免疫确认。最后，开发一种酶联免疫吸附测定（ELISA）试剂盒，并通过对456例BD临床样本的评估进行临床验证。
鉴定出一种分子量约为27 kDa的抗原为热休克蛋白27（HSP27）。在91例BD患者中的52例（57%）、92例类风湿关节炎（RA）患者中的66例（72%）、90例干燥综合征（SS）患者中的32例（36%）、92例系统性红斑狼疮（SLE）患者中的22例（24%）以及91例健康对照（HC）中的0例检测到血清IgG对重组人HSP27的反应性。BD血清IgG抗体对HSP27的反应性显著高于SLE（P<0.0001）、SS（P<0.0001）和HC（P<0.0001）。
本研究鉴定出HSP27为BD的候选内皮细胞自身抗原，这一发现很有趣且可能值得进一步探索。
