针对休眠结核分枝杆菌重组疫苗rBCG-Rv3133c的免疫效应研究  

Study on Immunogenicity Elicited by a Recombinant Vaccine of rBCG-Rv3133c to Fight Against Dormancy Mycobacterium tuberculosis

作　　者：邓仪昊[1] 何红云[1] 张本斯[1] 

机构地区：[1]大理学院基础医学院人体解剖学教研室,大理671000

出　　处：《生物医学工程学杂志》2013年第4期817-821,共5页Journal of Biomedical Engineering

基　　金：大理学院博士科研启动基金资助项目(KYBS201103)

摘　　要：为获取针对休眠期结核分枝杆菌(MTB)疫苗,本研究以编码MTB休眠相关转录调节蛋白DosR的基因Rv3133c为靶基因构建重组卡介苗,并研究其免疫效应。在本研究中,将从结核分枝杆菌H37Rv基因组获取的Rv3133c插入大肠杆菌-分枝杆菌穿梭表达质粒pMV361构建重组质粒,该重组体以电转化方式导入卡介苗基因组构建重组卡介苗rBCG-Rv3133c。将成功构建的重组卡介苗免疫BALB/c小鼠,分别在免疫后30d及180d,通过检测疫苗诱导的血清抗体滴度水平评价其诱导的体液免疫反应,通过检测免疫小鼠脾淋巴细胞受TB-PPD刺激后脾CD4+和CD8+T细胞的比例变化、细胞因子IFN-γ的表达评价重组疫苗诱导的细胞免疫。结果显示,重组疫苗rB-CG-Rv3133c可诱导产生较BCG更高水平的特异性抗体滴度、更强的CD4+和CD8+T细胞增殖反应及较高水平的IFN-γ表达量,提示该疫苗具有较强的研究应用价值。To obtain a vaccine to defend from dormancy Mycobacterium tuberculosis,we constructed the recombinant Bacilli Calmette-Guérin（BCG） vaccine with Rv3133c encoding dormancy-correlated transcriptional regulatory protein DosR in Mycobacterium tuberculosis as a target gene,and evaluated its immunogenicity in BALB/c mice.In this study,we constructed the recombinant plasmids of rpMV361-Rv3133c using gene colon technology.We then transformed BCG strains with above-mentioned plasmids to obtain recombinant vaccine of rBCG-Rv3133c.We used the rBCG strains successfully constructed to vaccinate in BALB/c mice.30d and 180d after immunization,the specific antibody titers were determined to investigate humoral responses induced by recombin ant vaccine.We detected changes of splenocyte subsets of CD4＋T、CD8＋ T cells and cytokine of IFN-γ secreted by splenocytes for evaluation of cellular immune responses.The results showed that the rBCG-Rv3133c was able to induce higher levels of antibody titer,stronger proliferative responses and higher IFN-γ production comparing with BCG vaccine.The results also suggested that this recombinant vaccine was a more efficacious tuberculosis vaccine for further study.

关 键 词：休眠结核分枝杆菌 Rv3133c 重组卡介苗 免疫效应 

分 类 号：R392.9[医药卫生—免疫学]