College of Life Science, Northwest Normal University, Lanzhou, 730070, Gansu, China.
Lanzhou Center for Tuberculosis Research and Gansu Provincial Key Laboratory of Evidence Based Medicine and Clinical Translation, Lanzhou University, Lanzhou, 730000, China.
Sci Rep. 2021 Jan 13;11(1):1249. doi: 10.1038/s41598-020-80899-6.
Tuberculosis (TB) kills more individuals in the world than any other disease, and a threat made direr by the coverage of drug-resistant strains of Mycobacterium tuberculosis (Mtb). Bacillus Calmette-Guérin (BCG) is the single TB vaccine licensed for use in human beings and effectively protects infants and children against severe military and meningeal TB. We applied advanced computational techniques to develop a universal TB vaccine. In the current study, we select the very conserved, experimentally confirmed Mtb antigens, including Rv2608, Rv2684, Rv3804c (Ag85A), and Rv0125 (Mtb32A) to design a novel multi-epitope subunit vaccine. By using the Immune Epitopes Database (IEDB), we predicted different B-cell and T-cell epitopes. An adjuvant (Griselimycin) was also added to vaccine construct to improve its immunogenicity. Bioinformatics tools were used to predict, refined, and validate the 3D structure and then docked with toll-like-receptor (TLR-3) using different servers. The constructed vaccine was used for further processing based on allergenicity, antigenicity, solubility, different physiochemical properties, and molecular docking scores. The in silico immune simulation results showed significant response for immune cells. For successful expression of the vaccine in E. coli, in-silico cloning and codon optimization were performed. This research also sets out a good signal for the design of a peptide-based tuberculosis vaccine. In conclusion, our findings show that the known multi-epitope vaccine may activate humoral and cellular immune responses and maybe a possible tuberculosis vaccine candidate. Therefore, more experimental validations should be exposed to it.
结核病（TB）是世界上导致死亡人数最多的疾病，而耐多药结核分枝杆菌（Mtb）的出现更是加剧了这一威胁。卡介苗（BCG）是唯一获准用于人类的结核病疫苗，能有效保护婴儿和儿童免受严重的军事和脑膜性结核病的侵害。我们应用先进的计算技术开发了一种通用的结核病疫苗。在当前的研究中，我们选择了非常保守、经过实验证实的 Mtb 抗原，包括 Rv2608、Rv2684、Rv3804c（Ag85A）和 Rv0125（Mtb32A），用于设计一种新型的多表位亚单位疫苗。我们使用免疫表位数据库（IEDB）预测了不同的 B 细胞和 T 细胞表位。还添加了一种佐剂（Griselimycin）来提高疫苗的免疫原性。使用生物信息学工具来预测、优化和验证 3D 结构，然后使用不同的服务器与 Toll 样受体（TLR-3）对接。根据过敏原性、抗原性、溶解性、不同的物理化学性质和分子对接分数，对构建的疫苗进行了进一步处理。基于计算机的免疫模拟结果显示对免疫细胞有显著的反应。为了使疫苗在大肠杆菌中成功表达，进行了计算机克隆和密码子优化。这项研究也为设计基于肽的结核病疫苗提供了一个良好的信号。总之，我们的研究结果表明，这种已知的多表位疫苗可能会激活体液和细胞免疫反应，可能成为一种潜在的结核病疫苗候选物。因此，应该对其进行更多的实验验证。
