Table_1_Bioinformatics analysis and consistency verification of a novel tuberculosis vaccine candidate HP13138PB.docx

BackgroundWith the increasing incidence of tuberculosis (TB) and the shortcomings of existing TB vaccines to prevent TB in adults, new TB vaccines need to be developed to address the complex TB epidemic. MethodThe dominant epitopes were screened from antigens to construct a novel epitope vaccine termed HP13138PB. The immune properties, structure, and function of HP13138PB were predicted and analyzed with bioinformatics and immunoinformatics. Then, the immune responses induced by the HP13138PB were confirmed by enzyme-linked immunospot assay (ELISPOT) and Th1/Th2/Th17 multi-cytokine detection kit. ResultThe HP13138PB vaccine consisted of 13 helper T lymphocytes (HTL) epitopes, 13 cytotoxic T lymphocytes (CTL) epitopes, and 8 B-cell epitopes. It was found that the antigenicity, immunogenicity, and solubility index of the HP13138PB vaccine were 0.87, 2.79, and 0.55, respectively. The secondary structure prediction indicated that the HP13138PB vaccine had 31% of α-helix, 11% of β-strand, and 56% of coil. The tertiary structure analysis suggested that the Z-score and the Favored region of the HP13138PB vaccine were -4.47 88.22%, respectively. Furthermore, the binding energies of the HP13138PB to toll-like receptor 2 (TLR2) was -1224.7 kcal/mol. The immunoinformatics and real-world experiments showed that the HP13138PB vaccine could induce an innate and adaptive immune response characterized by significantly higher levels of cytokines such as interferon-gamma (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and IL-10. ConclusionThe HP13138PB is a potential vaccine candidate to prevent TB, and this study preliminarily evaluated the ability of the HP13138PB to generate an immune response, providing a precursor target for developing TB vaccines.背景 随着结核病（Tuberculosis, TB）发病率的持续攀升，现有结核病疫苗在成人结核病预防中存在明显短板，亟需开发新型结核病疫苗以应对复杂的结核病流行形势。 方法 从抗原中筛选优势表位，构建一款命名为HP13138PB的新型表位疫苗。借助生物信息学与免疫信息学工具，预测并分析HP13138PB的免疫特性、结构与功能。随后通过酶联免疫斑点试验（enzyme-linked immunospot assay, ELISPOT）以及Th1/Th2/Th17多细胞因子检测试剂盒，验证HP13138PB诱导的免疫应答。 结果 HP13138PB疫苗包含13个辅助性T淋巴细胞（helper T lymphocytes, HTL）表位、13个细胞毒性T淋巴细胞（cytotoxic T lymphocytes, CTL）表位以及8个B细胞表位。经检测，HP13138PB的抗原性、免疫原性与可溶性指数分别为0.87、2.79与0.55。二级结构预测结果显示，HP13138PB的α螺旋占比31%、β折叠（β-strand）占比11%、无规卷曲（coil）占比56%。三级结构分析表明，HP13138PB的Z评分（Z-score）与拉马钱德兰图允许构象区域（Favored region）占比分别为-4.47与88.22%。此外，HP13138PB与Toll样受体2（toll-like receptor 2, TLR2）的结合能为-1224.7 kcal/mol。免疫信息学分析与实体实验结果均证实，HP13138PB可诱导先天免疫与适应性免疫应答，其特征为干扰素-γ（interferon-gamma, IFN-γ）、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素-4（interleukin-4, IL-4）以及IL-10等细胞因子水平显著升高。 结论 HP13138PB是一款极具应用潜力的结核病预防候选疫苗，本研究初步评估了HP13138PB诱导免疫应答的能力，为结核病疫苗的开发提供了潜在的前驱靶点。