表达HIV-1 Gag-Pol抗原的重组痘病毒的构建及免疫原性

目的构建稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体疫苗,并检测其体液免疫效果。方法将修饰型HIV-1Gag-Pol基因插入到痘病毒表达载体pSC11m2启动子P7·5下游,经与野生型痘病毒同源重组后,进行蓝白斑筛选。用筛选得到的重组病毒rM-GP转染HEK293细胞,经SDS-PAGE和Westernblot检测表达蛋白。用该重组病毒免疫BALB/c小鼠,并检测体液免疫应答。结果已筛选到稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体rM-GP,经Westernblot,在免疫小鼠血清中检测到抗HIV-1p24蛋白的特异性抗体。结论重组痘病毒载体rM-GP能稳定高效表达HIV-1Gag-Pol蛋白,并能引起有效的体液免疫应答。

Construction and Immunogenicity of Recombinant Modified Vaccinia Ankara Expressing HIV-1 Gag-Pol Antigen

Objective To construct the recombinant modified vacinia Ankara(rMVA)for stable and high expression of HIV-1 Gag-Pol antigen and study its humoral immune effect.Methods Insert the modified HIV-1 Gag-Pol gene downstream to the promoter P7.5 of expression vector pSC11m2 for homologous recombination with wild MVA,then select positive clones by white and blue screening.Transfect HEK293 cells with the obtained recombinant plasmid rM-GP and identify the expressed Gag-Pol protein by SDS-PAGE and Western blot.Immunize BALB/c mice with rM-GP and determine the humoral immune response by Western blot.Results rM-GP for stable and high expression of HIV-1 Gag-Pol was screened.Western blot showed specific antibody against HIV-1 p24 in the sera of mice immunized with rM-GP.Conclusion The rMVA for stable and high expression of HIV-1 Gag-Pol antigen was successfully constructed,and induced humoral immune response in mice.