Guo Fangzheng, Dong Sihang, Song Yamin, Qian Yuanyuan, Jiang Hairui, Zhang Wutong, Li Baiqing, Qian Zhongqing, Wang Xiaojing, Xu Guangxian, Wang Hongtao, Xu Tao
Anhui Province Key Laboratory of Immunology in Chronic Diseases, Laboratory Medicine Experimental Center, Laboratory Medicine College, Bengbu Medical University, Bengbu 233030, China.
Anhui Province Key Laboratory of Respiratory Tumor and Infectious Disease, Molecular Diagnosis Center, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China.
Int J Biol Macromol. 2025 Aug;320(Pt 4):146101. doi: 10.1016/j.ijbiomac.2025.146101. Epub 2025 Jul 17.
Tuberculosis (TB) is a chronic and persistent infectious disease caused by Mycobacterium tuberculosis (Mtb), posing a serious threat to global public health. The only widely available vaccine for TB is Bacillus Calmette-Guérin (BCG), and its limitations have become increasingly apparent in recent years, underscoring the urgent need for new TB vaccines. Moreover, subunit vaccines undergoing clinical trials face challenges such as complex production processes and weak immunogenicity. Efforts must be made to develop a safe, effective, easily mass-produced TB vaccine. Here, early secreted antigenic target of 6 kDa (ESAT6), culture filtrate protein of 10 kDa (CFP10) and ESAT6-CFP10 (EC) fusion epitopes from the Mtb region of difference (RD) were selected to construct traditional subunit antigens and self-assembling ferritin-based nanovaccines, EF, CF, and ECF. The transmission electron microscopy and dynamic light scattering showed that the three antigens coupled with ferritin self-assembled into uniform nanoparticles with even sizes. H&E staining demonstrated that the nanovaccines were safe. EF, CF, and ECF induced higher levels of cell proliferation than BCG and conventional subunit vaccines in immunized mice exposed to antigenic stimulation. TB nanovaccines enhanced the IFN-γ recall response and strongly stimulated the secretion of various TB-related cytokines, particularly Th1-type cytokines. Multi-color flow cytometry revealed that the nanovaccines, especially ECF, induced immune memory involving IFN-γT/T, CD4IL-2T and CD8IL-2T cells. Ferritin-NPs hold potential as a TB vaccine carrier and compared to BCG and traditional subunit vaccines, Ferritin-based TB nanoparticles produced by prokaryotic expression display superior immunogenicity.
结核病（TB）是由结核分枝杆菌（Mtb）引起的一种慢性持续性传染病，对全球公共卫生构成严重威胁。目前唯一广泛使用的结核病疫苗是卡介苗（BCG），其局限性近年来日益明显，凸显了开发新型结核病疫苗的迫切需求。此外，正在进行临床试验的亚单位疫苗面临着生产工艺复杂和免疫原性弱等挑战。必须努力开发一种安全、有效、易于大规模生产的结核病疫苗。在此，选择来自Mtb差异区域（RD）的早期分泌性6 kDa抗原靶标（ESAT6）、10 kDa培养滤液蛋白（CFP10）和ESAT6-CFP10（EC）融合表位来构建传统亚单位抗原和基于铁蛋白自组装的纳米疫苗EF、CF和ECF。透射电子显微镜和动态光散射显示，这三种与铁蛋白偶联的抗原自组装成大小均匀的纳米颗粒。苏木精-伊红染色表明纳米疫苗是安全的。在接受抗原刺激的免疫小鼠中，EF、CF和ECF诱导的细胞增殖水平高于卡介苗和传统亚单位疫苗。结核病纳米疫苗增强了IFN-γ回忆反应，并强烈刺激了各种结核病相关细胞因子的分泌，尤其是Th1型细胞因子。多色流式细胞术显示，纳米疫苗，尤其是ECF，诱导了涉及IFN-γT/T、CD4IL-2T和CD8IL-2T细胞的免疫记忆。铁蛋白纳米颗粒作为结核病疫苗载体具有潜力，与卡介苗和传统亚单位疫苗相比，原核表达产生的基于铁蛋白的结核病纳米颗粒具有更强的免疫原性。
