针对结核分枝杆菌潜伏感染的DNA疫苗V569免疫原性及保护性的初步研究  

Immunogenicity and protective results of DNA vaccine V569 against latent tuberculosis infection in animal models

作　　者：粟海波 刘梓健 周洋洋 彭宝洲 龚青[1] SU Haibo;LIU Zijian;ZHOU Yangyang;PENG Baozhou;GONG Qing(GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]广州医科大学广州医科大学-中国科学院广州生物医药与健康研究院联合生命科学学院

出　　处：《微生物与感染》2019年第5期289-296,共8页Journal of Microbes and Infections

基　　金：广东省高校优秀青年创新人才培养计划资助项目(2016KQNCX141);广州市科技计划一般项目(201804010317);广东省自然科学基金面上项目(2018A030313560)

摘　　要：为研究针对结核分枝杆菌潜伏感染的DNA疫苗,基于质粒A39构建了p-VAX1-Ag85B-Rv3425-Rv2029c-PPE26(V569)质粒DNA,并对其免疫原性及保护性进行初步研究。免疫性评价试验共分6组:PBS、p-VAX1-Ag85B(A)、p-VAX1-Ag85B-Rv3425(A3)、A39、V569和BCG,采用左后腿肌内注射C57BL/6小鼠,用流式细胞术和酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)分别检测细胞免疫和体液免疫水平;构建斑马鱼-海分枝杆菌潜伏感染模型,将PBS、A、A3、A39、BCG、V569分别通过腹腔注射免疫斑马鱼后,每日注射地塞米松10ug诱导海分枝杆菌复发感染,对斑马鱼肝脏进行菌落计数并绘制生存曲线。结果显示,与BCG组相比,V569能引发实验小鼠强烈的细胞免疫反应(IFN-γ高水平分泌),外周血CD4+/CD8+T细胞比例明显增加。在斑马鱼-海分枝杆菌潜伏感染复发模型中,与BCG免疫组相比,V569免疫斑马鱼后可显著减少其肝脏中海分枝杆菌数量,斑马鱼存活情况得到显著改善,表明V569 DNA疫苗可能是一种抗结核潜伏感染的候选DNA疫苗。We constructed the DNA vaccine Ag85B-Rv3425-Rv2029c-PPE26(V569)plasmid based on p-VAX1-Ag85B-Rv3425-Rv2029c(A39)and evaluated its immunogenicity in mice.The levels of cellular immunity and humoral immunity against PBS,BCG,p-VAX1-Ag85B(A),p-VAX1-Ag85B-Rv3425(A3),A39 and V569 were detected by enzyme-linked immunosorbent assay(ELISA)and flow cytometry after vaccination in mice.We found that V569 could trigger stronger Th1 immune response by augmenting the secretion of IFN-γand significantly improve the CD4+/CD8+T cell ratio in immunized mice compared to BCG.In addition,we evaluated the V569 as post-exposure DNA vaccine and found that it reduced bacterial burdens upon reactivation compared to BCG.The protection of V569 against latent TB infection was determined by zebrafish-Mycobacterium marinum latent infection model.In conclusion,V569 DNA vaccine may be a potential candidate DNA vaccine against latent tuberculosis infection.

关 键 词：PPE26 结核潜伏感染 DNA疫苗 Rv3425 Th1型细胞免疫反应 

分 类 号：R73[医药卫生—肿瘤]