NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Institute of Deep-sea Science and Engineering, Chinese Academy of Sciences, Sanya, Hainan, China.
Sci Rep. 2019 Feb 28;9(1):3042. doi: 10.1038/s41598-019-39402-z.
Tuberculosis (TB) is a continuing major threat to global health and a leading cause of death, particularly in developing countries. In this study, we aimed to identify a specific and sensitive diagnostic biomarker and develop a vaccine to prevent this disease. We investigated membrane proteins to reveal biomarkers in serum and peripheral blood mononuclear cells (PBMCs) obtained from TB patients. We employed Western blotting to evaluate serological immunoglobulin G levels, and Enzyme Linked Immunospot (ELISpot) to assess the antigen-specific cellular interferon-γ secretion from PBMCs after membrane protein stimulation. A total of 219 membrane proteins were identified, 52 exhibited at a higher levels than the 38-kDa prositive control. Of these 18 exhibited reacted ratios above 1, especially Rv1111c (427-981), with a ratios at 3.38. Accuracy and sensitivity were markedly higher for the top two antigen candidates, Rv0232 and Rv1115, after two rounds of ELISpot tests than ESAT-6 in the commercial kit (42.15 and 43.62%, respectively). These two proteins were administered to mice to detect whether they acted as effective antigens in vivo. These data provide a comprehensive view of the membranes involved in humoural and cellular immune responses that may be used as biomarkers for TB and candidates for a vaccine.
结核病（TB）是对全球健康的持续重大威胁，也是导致死亡的主要原因，特别是在发展中国家。在这项研究中，我们旨在确定一种特定且敏感的诊断生物标志物并开发一种预防这种疾病的疫苗。我们研究了膜蛋白，以揭示从结核病患者获得的血清和外周血单核细胞（PBMC）中的生物标志物。我们采用 Western blot 评估血清免疫球蛋白 G 水平，并用酶联免疫斑点（ELISpot）评估膜蛋白刺激后 PBMC 中抗原特异性细胞干扰素-γ的分泌。共鉴定出 219 种膜蛋白，其中 52 种的表达水平高于 38-kDa 阳性对照。其中 18 种的反应比值大于 1，尤其是 Rv1111c（427-981），比值为 3.38。在两轮 ELISpot 试验后，两种抗原候选物 Rv0232 和 Rv1115 的准确度和灵敏度明显高于商业试剂盒中的 ESAT-6（分别为 42.15%和 43.62%）。将这两种蛋白质施用于小鼠，以检测它们在体内是否作为有效抗原起作用。这些数据提供了涉及体液和细胞免疫反应的膜的全面视图，可作为结核病的生物标志物和疫苗候选物。
