HIV多表位核酸疫苗构建及其免疫原性

目的设计新型HIV复合多表位疫苗,并检测其在小鼠体内的免疫效果。方法检索抗原表位数据库,进行新型HIV多表位核酸疫苗的设计,利用化学合成的方法合成多表位基因,并构建重组质粒pVAX1-MEGNp24,转染BHK-21细胞,间接免疫荧光法检测多表位基因的表达。重组质粒免疫BALB/c小鼠,ELISA法检测抗体动态变化,流式细胞仪检测脾T淋巴细胞亚类。结果重组质粒pVAX1-MEGNp24经酶切及测序分析,证明构建正确。间接免疫荧光显示能在BHK-21细胞中表达多表位基因。免疫小鼠可诱导小鼠特异性体液免疫和细胞免疫。结论已成功构建了重组质粒pVAX1-MEGNp24,小鼠免疫试验显示其具有良好的免疫原性。

Construction and Immunogenicity of Multiple-epitope DNA Vaccine against HIV

Objective To design a novel multiple-epitope DNA vaccine against HIV and study its immunogenicity in mice.Methods The full-length of MEGN gene was synthesized according to the designed sequence by chemical method and used for the construction of recombinant plasmid pVAX1-MEGNp24.Transfect BHK-21 cells with the constructed recombinant plasmid and identify the expressed product by IFA.Immunize BALB/c mice with the recombinant plasmid and determine the serum antibody by ELISA,and the T lymphocyte subgroup by flow cytometry.Results Restriction analysis and DNA sequencing proved that recombinant plasmid pVAX1-MEGNp24 was successfully constructed.IFA showed that the constructed recombinant plasmid was expressed in BHK-21 cells.Both humoral and cellular immunities were induced in mice,as proved by ELISA and flow cytometry respectively.Conclusion Recombinant plasmid pVAX1-MEGNp24 as a multiple-epitope DNA vaccine against HIV was successfully constructed and showed good immunogenicity in mice.