Zheng Jianhua, Chen Lihong, Liu Liguo, Li Haifeng, Liu Bo, Zheng Dandan, Liu Tao, Dong Jie, Sun Lilian, Zhu Yafang, Yang Jian, Zhang Xiaobing, Jin Qi
‡From the MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Centre for Tuberculosis, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
‡From the MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Centre for Tuberculosis, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Mol Cell Proteomics. 2017 Sep;16(9):1578-1590. doi: 10.1074/mcp.M116.065813. Epub 2017 Jul 21.
Tuberculosis (TB) is one of the leading causes of death worldwide, especially in developing countries. Neonatal BCG vaccination occurs in various regions, but the level of protection varies in different populations. Recently, is found to be an immunomodulating therapeutic agent that could confer a significant level of protection against TB. It is the only vaccine in a phase III trial from WHO to assess its efficacy and safety in preventing TB disease in people with latent TB infection. However, the mechanism of immunotherapy of remains poorly understood. In this study, the full genome of was obtained by next-generation sequencing technology, and a proteogenomic approach was successfully applied to further perform genome annotation using high resolution and high accuracy MS data. A total of 3,387 proteins (22,508 unique peptides) were identified, and 581 proteins annotated as hypothetical proteins in the genome database were confirmed. Furthermore, 38 novel protein products not annotated at the genome level were detected and validated. Additionally, the translational start sites of 445 proteins were confirmed, and 98 proteins were validated through extension of their translational start sites based on N terminus-derived peptides. The physicochemical characteristics of the identified proteins were determined. Thirty-five immunogenic proteins of were identified by immunoproteomic analysis, and 20 of them were selected to be expressed and validated by Western blot for immunoreactivity to serum from patients infected with The results revealed that eight of them showed strong specific reactive signals on the immunoblots. Furthermore, cellular immune response was further examined and one protein displayed a higher cellular immune level in pulmonary TB patients. Twelve identified immunogenic proteins have orthologous in H37Rv and BCG. This is the first study to obtain the full genome and annotation of using a proteogenomic approach, and some immunogenic proteins that were validated by immunoproteomic analysis could contribute to the understanding of the mechanism of immunotherapy.
结核病（TB）是全球主要死因之一，在发展中国家尤为如此。新生儿卡介苗接种在不同地区开展，但不同人群中的保护水平存在差异。最近，[具体物质未提及]被发现是一种免疫调节治疗剂，可提供对结核病的显著保护水平。它是世界卫生组织进行III期试验以评估其在预防潜伏性结核感染人群中结核病的疗效和安全性的唯一疫苗。然而，[具体物质未提及]免疫治疗的机制仍知之甚少。在本研究中，通过下一代测序技术获得了[具体物质未提及]的全基因组，并成功应用蛋白质基因组学方法，利用高分辨率和高精度的质谱数据进一步进行基因组注释。共鉴定出3387种蛋白质（22508个独特肽段），并确认了基因组数据库中注释为假设蛋白质的581种蛋白质。此外，检测并验证了38种在基因组水平未注释的新型蛋白质产物。另外，确认了445种蛋白质的翻译起始位点，并基于N端衍生肽通过延长其翻译起始位点对98种蛋白质进行了验证。确定了所鉴定蛋白质的理化特性。通过免疫蛋白质组学分析鉴定出[具体物质未提及]的35种免疫原性蛋白质，其中20种被选择表达并通过蛋白质印迹法验证其对感染[具体物质未提及]患者血清的免疫反应性。结果显示，其中8种在免疫印迹上显示出强烈的特异性反应信号。此外，进一步检测了细胞免疫反应，一种蛋白质在肺结核患者中显示出较高的细胞免疫水平。12种鉴定出的免疫原性蛋白质在H37Rv和卡介苗中有直系同源物。这是首次使用蛋白质基因组学方法获得[具体物质未提及]的全基因组和注释，一些经免疫蛋白质组学分析验证的免疫原性蛋白质可能有助于理解[具体物质未提及]免疫治疗的机制。
