Table_2_Evaluation of the consistence between the results of immunoinformatics predictions and real-world animal experiments of a new tuberculosis vaccine MP3RT.doc

BackgroundOur previous study developed a novel peptide-based vaccine, MP3RT, to fight against tuberculosis (TB) infection in a mouse model. However, the consistency between the immunoinformatics predictions and the results of real-world animal experiments on the MP3RT vaccine remains unclear.
MethodIn this study, we predicted the antigenicity, immunogenicity, physicochemical parameters, secondary structure, and tertiary structure of MP3RT using bioinformatics technologies. The immune response properties of the MP3RT vaccine were then predicted using the C-ImmSim server. Finally, humanized mice were used to verify the characteristics of the humoral and cellular immune responses induced by the MP3RT vaccine.
ResultsMP3RT is a non-toxic and non-allergenic vaccine with an antigenicity index of 0.88 and an immunogenicity index of 0.61, respectively. Our results showed that the MP3RT vaccine contained 53.36% α-helix in the secondary structure, and the favored region accounted for 98.22% in the optimized tertiary structure. The binding affinities of the MP3RT vaccine to the human leukocyte antigen (HLA)-DRB1*01:01 allele, toll-like receptor-2 (TLR-2), and TLR-4 receptors were -1234.1 kcal/mol, -1066.4 kcal/mol, and -1250.4 kcal/mol, respectively. The results of the C-ImmSim server showed that the MP3RT vaccine could stimulate T and B cells to produce immune responses, such as high levels of IgM and IgG antibodies, IFN-γ, TNF-α, and IL-2 cytokines. Results from real-world animal experiments showed that the MP3RT vaccine could stimulate the humanized mice to produce high levels of IgG and IgG2a antibodies and IFN-γ+ T lymphocytes. Furthermore, the levels of IFN-γ, IL-2, and IL-6 cytokines in mice immunized with the MP3RT vaccine were significantly higher than those in the control group.
ConclusionMP3RT is a highly antigenic and immunogenic potential vaccine that can effectively induce Th1-type immune responses in silico analysis and animal experiments. This study lays the foundation for evaluating the value of computational tools and immunoinformatic techniques in reverse vaccinology research.

背景：本团队前期研究开发了一种新型肽类疫苗MP3RT，可在小鼠模型中对抗结核病（tuberculosis, TB）感染。然而，该疫苗的免疫信息学（immunoinformatics）预测结果与真实动物实验结果之间的一致性仍未明确。
方法：本研究利用生物信息学（bioinformatics）技术预测了MP3RT的抗原性、免疫原性、理化参数、二级结构与三级结构；随后通过C-ImmSim服务器预测该疫苗的免疫应答特性；最终使用人源化小鼠验证MP3RT诱导的体液免疫与细胞免疫应答特征。
结果：MP3RT是一种无毒、无致敏性的疫苗，其抗原性指数为0.88，免疫原性指数为0.61。研究结果显示，MP3RT的二级结构中α-螺旋占比达53.36%；优化后的三级结构中，偏好区域占比为98.22%。该疫苗与人类白细胞抗原（HLA）-DRB1*01:01等位基因、Toll样受体2（TLR-2）及TLR-4受体的结合亲和力分别为-1234.1 kcal/mol、-1066.4 kcal/mol与-1250.4 kcal/mol。C-ImmSim服务器的预测结果表明，MP3RT可刺激T细胞与B细胞产生免疫应答，如高水平的免疫球蛋白M（IgM）、免疫球蛋白G（IgG）抗体以及干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）和白细胞介素-2（IL-2）等细胞因子。真实动物实验结果显示，MP3RT可刺激人源化小鼠产生高水平的IgG、IgG2a抗体以及IFN-γ阳性T淋巴细胞；此外，免疫接种MP3RT的小鼠体内IFN-γ、IL-2及IL-6细胞因子水平显著高于对照组。
结论：MP3RT是一种高抗原性、高免疫原性的候选疫苗，可在虚拟分析与动物实验中有效诱导Th1型免疫应答。本研究为评估计算工具与免疫信息学技术在反向疫苗学（reverse vaccinology）研究中的应用价值奠定了基础。