Peng Chen1, Lili Shi1, Yun Jiang1, Yuting Ji1, Hai Yan1, Shutao Sun2, Yiping Xun1, Guangyu Chen3, Xiaoxu Wang1, Weiyang Chen1, Hongwu Du4. 1. School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China. 2. Core Facility, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. 3. ImmunoHunt Corporation, 139 Fengtai Rd, Beijing 100071, China. 4. School of Chemistry and Biotechnology Engineering, University of Science & Technology Beijing, Beijing 100083, China. Electronic address: hongwudu@ustb.edu.cn.
Abstract
OBJECTIVE: The aim of this study was to identify candidate pathogenic autoantigens of Behçet's disease (BD) in pathogen-stimulated target cells. METHODS: 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. RESULTS: 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). CONCLUSIONS: This study identified HSP27 as a candidate endothelial cell autoantigen of BD, which is interesting and probably worth further exploration.
OBJECTIVE: The aim of this study was to identify candidate pathogenic autoantigens of Behçet's disease (BD) in pathogen-stimulated target cells. METHODS: 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. RESULTS: 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 humanHSP27 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). CONCLUSIONS: This study identified HSP27 as a candidate endothelial cell autoantigen of BD, which is interesting and probably worth further exploration.