Chen Peng, Zhang Chunyan, Liu Chen, Zhang Liyun, Yang Chunhe, Chen Guangyu, Ma Dan, Tian Yaping, Du Hongwu
112 Lab, School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China.
Department of Clinical Biochemistry, Chinese PLA General Hospital, Beijing 100853, China.
EBioMedicine. 2016 Apr;6:215-221. doi: 10.1016/j.ebiom.2016.03.009. Epub 2016 Mar 10.
The aim of this study was to verify the hypothesis originated from bioinformatics and literature reviews that hnNRP A1 may be a new immune target of Behçet's disease (BD).
First, bioinformatics was used to show the correlation between hnRNP A1 and A2/B1 in amino acid sequences and three dimensional structures. Second, hnRNP A1 was expressed, purified, and immunologically confirmed by systematic immunology methods including: Western blotting, immunoprecipitation and Dot-ELISA. Then, ELISA was used to screen the anti-hnRNP A1 autoantibodies in newly confirmed clinical samples and the clinical significance was compared between anti-hnRNP A1 antibody positive and negative groups. Finally, the endothelial cells antigen profile of one anti-hnRNP A1 antibody positive BD patient was detected using immunoprecipitation with liquid chromatography tandem mass spectrometry (LC-TMS).
In total 720 subjects enrolled and tested in this study. Our results demonstrated hnRNP A1 as a new immune target of BD. The reactivity of BD serum IgG antibodies against hnRNP A1 was significantly higher than healthy controls (P<0.0001), and deep vein thrombosis (DVT) showed a significant higher in the anti-hnRNP A1 antibodies positive group (P<0.05).
本研究旨在验证源自生物信息学和文献综述的假说，即hnNRP A1可能是白塞病（BD）的一个新的免疫靶点。
首先，利用生物信息学展示hnRNP A1与A2/B1在氨基酸序列和三维结构上的相关性。其次，表达、纯化hnRNP A1，并通过包括蛋白质印迹法、免疫沉淀法和斑点酶联免疫吸附测定在内的系统免疫学方法进行免疫确认。然后，采用酶联免疫吸附测定法筛选新确诊临床样本中的抗hnRNP A1自身抗体，并比较抗hnRNP A1抗体阳性和阴性组之间的临床意义。最后，使用液相色谱串联质谱免疫沉淀法（LC-TMS）检测一名抗hnRNP A1抗体阳性BD患者的内皮细胞抗原谱。
本研究共纳入并检测了720名受试者。我们的结果证明hnRNP A1是BD的一个新的免疫靶点。BD血清IgG抗体对hnRNP A1的反应性显著高于健康对照（P<0.0001），且抗hnRNP A1抗体阳性组的深静脉血栓形成（DVT）发生率显著更高（P<0.05）。
