共刺激分子B7-1与汉坦病毒核蛋白基因共表达载体的构建及免疫调节作用

目的:探讨共刺激分子B7-1(CD80)对汉坦病毒核蛋白基因疫苗的免疫调节作用.方法:构建双启动子共表达B7-1基因和汉坦病毒核蛋白基因的真核表达载体pcDNA 3.1-B7-S,酶切鉴定后直接肌注免疫BALB/c小鼠,同时设对照组pcDNA 3.1 空质粒接种组、pcDNA 3.1-S接种组.ELISA法检测血清特异性抗体,MTT法检测T细胞增殖反应.结果:pcDNA 3.1-B7-S接种组鼠在抗体的产生水平及淋巴细胞增殖指数上均较pcDNA 3.1-S接种组明显增高.结论:接种B7-1基因和汉坦病毒核蛋白基因的共表达质粒优于注射单目的抗原基因表达质粒,为探索增强基因疫苗的免疫作用提供了新的途径.

Construction and immune regulation of the co-expressing vector of costimulation molecules B7-1 and nucleiocapsid protein derived from hantavirus

Objective: To study the effect of costimulation molecules B7-1(CD80)on the immune responses induced by recombinant plasmid encoding nucleiocapsid protein derived from hantavirus. Methods: BALB/c mice were immunized 3 times through intramuscular injection of recombinant plasmid co-expressing of B7-1 costimulation molecules and nucleiocapsid protein derived from hantavirus(pcDNA3.1-B7-S),which was verified by enzyme analysis. Meanwhile, pcDNA3.1+ and pcDNA3.1-S were as controls. To evaluate the humoral and cellular immune responses, antigen-specific lymphocyte proliferation and antibody production were assayed by using MTT method and ELISA respectively. Results: The mice immunized with pcDNA3.1-B7-S had higher level of antigen -specific IgG and lymphocyte proliferation than the group immunized with pcDNA3.1-S. Conclusion: The recombinant plasmid co-expressing of B7-1 costimulation molecules and nucleiocapsid protein derived from hantavirus can effectively improve the immunogenicity and efficacy, which provide a new vaccination strategy for enhancing the immune responses induced by a DNA vaccine.