摘要
目的 探索增强结核杆菌DNA疫苗有效性的新方法 ,为研制并开发新一代高效结核杆菌DNA疫苗奠定基础。方法 分别构建结核杆菌Ag85A抗原基因真核表达质粒及其与小鼠粒细胞 巨噬细胞集落刺激因子的真核嵌合表达质粒 ,体外转染COS7细胞检测两种重组质粒的表达活性后 ,将其分别用作DNA疫苗给BALB/c小鼠肌内注射 ,检测小鼠体内特异性体液免疫和细胞免疫应答相关指标 ,同时进行动物免疫保护性实验 ,比较分析两种DNA疫苗在小鼠体内的免疫效应。结果 结核杆菌Ag85A抗原蛋白基因与小鼠粒细胞 巨噬细胞集落刺激因子的嵌合DNA疫苗在小鼠体内的免疫原性明显强于Ag85A抗原基因非嵌合DNA疫苗 ,但两种DNA疫苗的免疫保护性无明显差异。结论 粒细胞 巨噬细胞集落刺激因子与结核杆菌免疫保护性抗原基因嵌合DNA疫苗能显著增强结核病DNA疫苗的免疫原性。
Objective To explore new method for enhancing the efficacy of tuberculosis DNA vaccine. Methods Two recombinant plasmids were constructed, one named as pBK GM/85A encoding mouse granulocyte macrophage colony stimulating factor(GM CSF) fused to Mycobacterium tuberculosis Ag85A antigen, the other named as pBK 85A encoding Mycobacterium tuberculosis Ag85A antigen alone. Subsequently, the two plasmids were transferred into cultured COS7 cells by using cationic liposomes. The expression products were identified by Western blotting. Then, in a murine model, we compared the immunogenicity and protective immunity of the two recombinant plasmids following genetic immunization. Results All pBK GM/85A injected mice elicited higher antibody titres than that for pBK 85A injected mice. Lymphocytes obtained from the spleen of pBK GM/85A immunized mice exhibited higher lymphocyte proliferative response、IFN γ production and CTL activity than that for pBK 85A immunized mice. The protective efficacy was also higher for pBK GM/85A immunized mice than that for pBK 85A immunized mice. However, The protective efficacy for pBK GM/85A immunized mice was lower than that for BCG immunized mice. Conclusion These results showed that DNA vaccines with GM CSF/antigen fusion constructs could greatly improve the immunogenicity of DNA vaccine against Mycobacterium tuberculosis.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2004年第6期386-390,共5页
Chinese Journal of Infectious Diseases
基金
四川省科委应用基础研究基金 (G0 10 2 0 )
