Tan Kun, Zhang Jingyan, Teng Xindong, Liang Jinping, Wang Xiaochun, Yuan Xuefeng, Tian Maopeng, Fan Xionglin
Department of Pathogen Biology, School of Basic Medicine, Huazhong University of Science & Technology, Wuhan 430030, PR China.
Department of Pathogen Biology, School of Basic Medicine, Huazhong University of Science & Technology, Wuhan 430030, PR China.
J Microbiol Methods. 2015 Jul;114:57-61. doi: 10.1016/j.mimet.2015.05.007. Epub 2015 May 7.
More efficacious and specific biomarkers are urgently needed for better control of tuberculosis (TB), the second leading infectious cause of mortality worldwide. The region of difference 9 (RD9) presents the genome of the causative pathogen Mycobacterium tuberculosis rather than other species of the genus Mycobacterium, which might be promising targets for specific diagnosis, vaccine development and pathogenesis. In this study, two proteins Rv2073c and Rv2074, encoded by the RD9 were expressed and purified from Escherichia coli system. Following stimulation with both proteins, the levels of IFN-γ secreted by T cells from a total of 49 whole blood samples obtained from clinically diagnosed active TB patients, patients with latent TB infections (LTBIs), and healthy donors, were compared with those of the incubation with recombinant fusion protein of CFP21 and MPT64 (rCM). Our results demonstrated that only Rv2073c could induce a higher level of IFN-γ in TB infections than healthy controls and there was a positive correlation between Rv2073c- and rCM-specific IFN-γ levels in TB infections and healthy donors, respectively. These findings indicate that Rv2073c might be a promising antigen for specific diagnostic reagents and vaccine candidates of TB.
为了更好地控制结核病(TB),全球第二大致死性传染病,迫切需要更有效、更具特异性的生物标志物。差异区域9(RD9)呈现了致病病原体结核分枝杆菌的基因组,而非分枝杆菌属的其他物种,这可能是特异性诊断、疫苗开发和发病机制研究的有前景的靶点。在本研究中,由RD9编码的两种蛋白质Rv2073c和Rv2074在大肠杆菌系统中表达并纯化。用这两种蛋白质刺激后,比较了从临床诊断的活动性结核病患者、潜伏性结核感染(LTBI)患者和健康供体获得的总共49份全血样本中T细胞分泌的IFN-γ水平与用CFP21和MPT64重组融合蛋白(rCM)孵育后的水平。我们的结果表明,只有Rv2073c能在结核病感染中诱导比健康对照更高水平的IFN-γ,并且在结核病感染和健康供体中,Rv2073c特异性和rCM特异性IFN-γ水平之间分别存在正相关。这些发现表明,Rv2073c可能是结核病特异性诊断试剂和候选疫苗的有前景的抗原。