摘要
目的构建包含结核杆菌3种抗原蛋白Ag85A、ESAT-6和CFP-10真核表达质粒的多价DNA疫苗,并对其在小鼠体内的免疫原性进行初步分析。方法PCR扩增获得结核杆菌ag85 a、esat-6和cfp-10基因片段,分别克隆入真核表达质粒VR1020的BamHⅠ位点构建DNA疫苗,以此多价DNA疫苗免疫BALB/c小鼠,检测小鼠体内的特异性抗体应答和细胞免疫反应,并与BCG免疫组、质粒VR1020免疫组和生理盐水对照组相比较。结果多价DNA疫苗免疫小鼠后能产生特异性的细胞和体液免疫应答,DNA疫苗免疫组小鼠脾细胞IFN-γ的表达量显著高于BCG免疫组,而DNA疫苗免疫组小鼠所产生的特异性抗体水平低于BCG免疫组。结论成功构建了结核杆菌多价DNA疫苗,免疫小鼠后检测到特异性细胞和体液免疫应答。
Objective To construct a multivalcnt DNA vaccine encoding Ag85A,ESAT-6 and CFP-10 antigens from Mycobacterium tuberculosis, and to preliminarily study its immunogcnicity. Methods Mycobacterium tuberculosis ag85a,csat-6 and cfp-10 gone fragments were obtained by using Polymcrasc Chain Reaction （PCR）. The gone fragments were cloned into the BamH I site of plasmid VRI020 to construct three recombinant plasmids （multivalcnt DNA vaccine）. BALB/c mice were immunized with the multivalcnt DNA vaccine,plasmid vector VRI020 or BCG. The hormonal immune response and cell-mediated immune response of the mice were detected and analyzed. Results Specific hormonal immune response and cell-mediated immune response were detected in multivalcnt DNA vaccine immunized mice and BCG immunized mice. The IFN-T production of lymphocytcs obtained from the multivalcnt DNA vaccine immunized mice were significantly higher than that from BCG immunized mice, while the specific antibody level of the multivalcnt DNA vaccine immunized mice was lower than that of BCG immunized mice. Conclusion A multivalcnt DNA vaccine against Mycobactcrium tuberculosis was constructed successfully. BALB/c mice immunized with this kind of DNA vaccine can induce both the hormonal immune response and the cell-mediated immune response.
出处
《川北医学院学报》
CAS
2007年第1期7-10,共4页
Journal of North Sichuan Medical College
基金
教育部重点项目(编号205138)
四川省教育厅重点项目(编号:2003A067)
关键词
结核杆菌
多价核酸疫苗
免疫原性
Mycobacterinm tuberculosis
DNA vaccine
Immunogenicity
