【摘要】 目的 为提高对成年人的保护效果，开发一种基于M.tb中AhpC抗原的新型结核病亚单位疫苗。方法 利用基因工程技术构建pET28a-AhpC重组质粒，在大肠杆菌中表达并纯化AhpC蛋白；应用WBIA和MFCIA检测AhpC抗原能否被M.tb感染者的外周血T细胞识别；AhpC/IC31疫苗诱导的免疫应答通过小鼠免疫实验进行评估，并通过ELISA和细胞因子检测进一步评估该疫苗的免疫原性和保护潜力。结果 成功构建并表达了重组AhpC抗原；rAhpC在M.tb感染者外周血中诱导了显著高水平的INF-γ、IL-6和TNF-α反应；AhpC/IC31成功诱导小鼠产生强烈的Th1型免疫应答，且在BCG初免后予以增强免疫，可刺激更高的免疫应答水平和保护潜力。结论 本研究提供了一种新型TB亚单位疫苗的候选靶抗原，为预防成人结核病、保护M.tb潜伏感染者提供了理论和实验依据。更多还原

【Abstract】 Objective Tuberculosis(TB) is a significant global public health issue, and the current vaccine, Bacillus Calmette-Guérin(BCG),offers limited protection for adults.This study aims to develop a novel TB subunit vaccine based on the AhpC antigen from Mycobacterium tuberculosis(M.tb) to enhance protection for adults.Methods The pET28a-AhpC recombinant plasmid was constructed by using genetic engineering techniques, and the AhpC protein was expressed and purified in E.coli.The immunogenicity of the AhpC antigen to be recognized by T cells in the peripheral blood of M.tb-infected individuals was detected by using Western Blot Immunoassay(WBIA) and Multiplex Fluoro-Cytometric Immunoassay(MFCIA).Further, the immunization experiment in mice was conducted to evaluate the immune response induced by the AhpC/IC31 vaccine, and its immunogenicity and protective potential were assessed through ELISA and cytokine detection.Results The recombinant AhpC antigen was successfully constructed and expressed.The rAhpC induced a significantly high level of INF-γ,IL-6 and TNF-α responses in the peripheral blood of M.tb-infected individuals.AhpC/IC31 successfully induced a strong Th1-type immune response in mice, which was enhanced after initial vaccination with BCG,leading to a higher level of immune response and protective potential.Conclusion This study provides a candidate target antigen for a new subunit vaccine against TB,offering a theoretical and experimental foundation for the prevention of adult tuberculosis and the protection of individuals with latent M.tb infection.更多还原