共表达结核杆菌Ag85A-ESAT-6融合抗原与IL-21核酸疫苗的构建及免疫效果研究  

Constructing Nucleic Acid Vaccine Expressing Fusion Antigens of Mycobacterium tuberculosis Ag85A and ESAT-6,and Interleukin 21 as well as Studying its Immune Effect in Mice

作　　者：李金龙[1] 余方流[1] 王亚琴[1] 羊海涛[2] 刘春生[1] 许卫国[2] 唐权[1] 何向峰[1] 蒋翠莲[1] 窦骏[1] 

机构地区：[1]东南大学基础医学院病原生物学与免疫学系,江苏南京210009 [2]江苏省疾病预防与控制中心,江苏南京210008

出　　处：《生物技术通讯》2009年第6期765-768,共4页Letters in Biotechnology

基　　金：江苏省自然科学基金重点项目(BK2007710)

摘　　要：目的:构建结核杆菌Ag85A-ESAT-6融合抗原与细胞因子IL-21共表达核酸疫苗pIRES-IL-21-Ag85A-ESAT-6,研究其对小鼠免疫功能的影响。方法:用PCR方法分别扩增出Ag85A和IL-21编码基因,并先后插入质粒pIRES-ESAT-6中,构成共表达核酸疫苗pIRES-IL-21-Ag85A-ESAT-6;免疫小鼠后,通过CTL和NK细胞杀伤活性和脾细胞增殖试验,以及小鼠血清抗体、IFN-γ和IL-4的检测,评价该核酸疫苗的免疫效果。结果:pIRES-IL-21-Ag85A-ESAT-6核酸疫苗免疫鼠的CTL活性、脾细胞增殖反应、IFN-γ水平及血清抗体产生明显高于对照组,差异有显著性意义。结论:pIRES-IL-21-Ag85A-ESAT-6核酸疫苗能够诱导小鼠产生有效的免疫反应,可作为新型结核疫苗的候选组分。Objective： To construct nucleic acid vaccine plRES-IL21-Ag85A-ESAT-6 expressing fusion antigens of Ag85A and ESAT-6 as well as IL-21 and to study its immunol effects. Methods： The genes of Ag85A and IL-21 were respectively amplified from the recombinant plasmid plRES-Ag85A and pRSC-IL-21 by PCR and then cloned directly into the recombinant plRES-ESAT-6, and formed the nucleic acid vaccine pIRES-IL-21-Ag85A-ESAT-6, then inoculating it into C57BL/6 mouse. The activities of cytotoxic T lymphocytes（CTL）, natural killer（NK） cells and spleen cells proliferation responses to purified protein derivative（PPD） were measured by MTT method in immunized mice respectively. The antibody and cytokine levels of serum were also detected by ELISA respectively. Results： The activity of CTL and spleen cell pro- liferation responses to PPD, IFN-γ level and its titer of antibody in mice immunized with the plRES-IL21-Ag85A-ESAT- 6 were significantly increased respectively compared with the control mice immunized with blank plasmid. Conclusion： The nucleic acid vaccine pIRES-IL-21-Ag85A-ESAT-6 can induce mice to generate effective immune response in mice model and it may conduct one of a new type candidate tuberculosis vaccine.

关 键 词：结核杆菌 AG85A 白细胞介素-21 ESAT-6 核酸疫苗 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]