HIV多表位核酸疫苗构建及其免疫原性  

Construction and Immunogenicity of Multiple-epitope DNA Vaccine against HIV

作　　者：李臻[1] 金宁一[2] 金洪涛[2] 沈国顺[3] 付延军[4] 辛苏文[5] 韩贞珍[4] 刘玉生[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军事医学科学院军事兽医研究所病毒室,长春130062 [3]沈阳农业大学畜牧兽医学院,沈阳110161 [4]延边大学农学院,龙井133400 [5]山西农业大学动物科技学院,太谷030801

出　　处：《中国生物制品学杂志》2007年第3期178-180,共3页Chinese Journal of Biologicals

基　　金：军队科技攻关重点项目(06G127);"863"计划项目(2003AA219051)

摘　　要：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。Objective To design a novel muhiple-epitope DNA vaccine against HIV and study its immunogenicity in mice. Methods The flail-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 pVAXI-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 Reeom- blnant plasmid pVAXI-MEGNp24 as a multiple-epitope DNA vaccine against HIV was successfully constructed and showed good immunogenieity in mice.

关 键 词：人免疫缺陷病毒 核酸疫苗 多表位抗原 

分 类 号：R373.9[医药卫生—病原生物学]