摘要
目的?构建结核杆菌ESAT6抗原DNA疫苗,并观察其在BALB/c小鼠体内诱导的免疫应答。方法:构建结核杆菌 ESAT6抗原DNA疫苗pVAX-ESAT6,并将其肌肉注射免疫BALB/c雌性小鼠,用ELISA方法检测小鼠的血清抗体(IgG、 IgG2a/IgG1)水平,并在体外用特异抗原刺激小鼠脾细胞,检测其分泌细胞因子(IFN-γ、IL-4)的水平;用乳酸脱氢酶(LDH)释 放法测定小鼠细胞毒性T淋巴细胞反应。结果:pVAX-ESAT6DNA疫苗诱导了抗原特异的体液免疫应答和细胞免疫应答, 抗体亚型检测显示IgG2a与IgG1的比值为(2.28±0.4),脾细胞分泌的细胞因子水平检测显示IFN-γ的水平(360±45pg/ml) 高于IL-4的水平(124±16pg/ml)。结论:成功地构建了pVAX-ESAT6DNA疫苗,它能在小鼠中诱导抗原特异的免疫应答, 应答类型以Th1型占优势。
Objective:To construct ESAT6DNA vaccine against Mycobacterium tuberculosis and to study the immune responses it induced in mice.Methods:DNA vaccine(pVAX-ESAT6)expressing ESAT6antigen was constructed and intra-muscularly inoculated into female BALB/c mice.The serum antibodies including IgG,IgG 2a /IgG 1 were detected by ELISA.The splenocytes from immunized mice were stimulated with ESAT6antigen in vitro and the secreted cytokines(IFN-γ,IL-4) were also assayed.The cytotoxic T lymphocyte response was determined by LDH release method.Results:The pVAX-ESAT6DNA vaccine induced antigen-specific humoral and cellular immune responses.The ratio of IgG 2a /IgG 1 was(2.28±0.4).IFN-γ[(360±45)pg/ml]from splenocytes was higher than that of IL-4[(124±16)pg/ml].Conclusion:pVAX-ESAT6DNA vaccine is successfully constructed,and it can induce antigen-specific humoral,with cellular immune responses and Th1type being the dominant. [
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2004年第6期642-645,共4页
Academic Journal of Second Military Medical University
基金
国家高新技术发展规划("863"计划)课题(2001AA213111).
