Liu Ying, Wu Lan, Zhou Ke-ming, Zhang Xu-dong, Hong Kun-sue, Shao Yi-ming
National Center for AIDS Prevention and Control. Center for Disease Control and Prevention, Beijing 100050, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2004 Jun;18(2):150-3.
Construction of replication-deficient recombinant adenovirus expressing gag-pol and env genes of human immunodeficiency virus (HIV) in mice.
gag-polDelta and gp140TM genes were cloned into shuttle vector pAdTrack-CMV respectively, and then the plasmids containing gag-polDelta or gp140TM gene were cotransformed with the backbone of adenovirus into E.coli BJ5183. Transfections of the recombinants were performed to obtain recombinant adenoviruses. Its immunogenicity was evaluated by testing antibody levels of mice primed with DNA vaccines and boosted with recombinant adenoviruses.
The replication-deficient recombinant adenovirus could express Gp140TM, Gag P55 and P24 proteins correctly. The mice primed with DNA vaccines and boosted with recombinant adenoviruses elicited high titer of HIV-1-specific antibody compared with that inoculated with DNA vaccines only.
Replication-deficient recombinant adenovirus expressing gag-polDelta and gp140TM can elicit high titer HIV-1-specific antibodies.
在小鼠体内构建表达人类免疫缺陷病毒(HIV)gag-pol和env基因的复制缺陷型重组腺病毒。
将gag-polDelta和gp140TM基因分别克隆到穿梭载体pAdTrack-CMV中,然后将含有gag-polDelta或gp140TM基因的质粒与腺病毒骨架共转化到大肠杆菌BJ5183中。对重组体进行转染以获得重组腺病毒。通过检测用DNA疫苗免疫并用重组腺病毒加强免疫的小鼠的抗体水平来评估其免疫原性。
复制缺陷型重组腺病毒能够正确表达Gp140TM、Gag P55和P24蛋白。与仅接种DNA疫苗的小鼠相比,用DNA疫苗免疫并用重组腺病毒加强免疫的小鼠产生了高滴度的HIV-1特异性抗体。
表达gag-polDelta和gp140TM的复制缺陷型重组腺病毒能够引发高滴度的HIV-1特异性抗体。