目的 了解pol基因修饰前后在不同载体系统中表达水平差异对免疫效果的影响,为确定HIV疫苗中能诱发较高水平细胞免疫的Pol靶抗原奠定实验基础.方法 将含有优化前后pol基因的HIV-1DNA( pVRC)疫苗和痘苗病毒载体(rVV)疫苗单独或联合免疫BALB/c小鼠,利用IFN -γ ELISPOT和ICS检测各组的细胞免疫效果,ELISA检测体液免疫水平,分别比较基因优化前后及在不同载体内的Pol诱发的免疫效果.结果 DNA疫苗中pol基因修饰后细胞免疫反应由112提高至258 (SFC/106SMNC),抗体滴度随免疫次数增加而提高,基因修饰后第二针的抗体水平由101.7提高至102.3,三针后则没有差别；而以痘苗病毒为载体的疫苗基因修饰前后细胞免疫反应( ～600SFC/106SMNC)和抗体水平(～102.2)均没有差异.两种疫苗联合免疫均可显著提高Pol在小鼠体内的免疫效果,基因修饰后细胞免疫反应由788提高至2500( SFC/106 SMNC),抗体水平则没有差别.结论 Pol基因修饰明显提高常规DNA疫苗免疫效果,并可进一步提高联合免疫效果,但对重组痘苗病毒疫苗单独免疫效果无明显影响.%Objective To understand the immune effect in different vectors after pol gene modification,study the Pol expression level differences on the immune response,to confirm the optimized pol may be good vaccine candidate antigen for use as a protective HIV vaccine based on the cellular mechanism.Methods BALB/c mice were injected with HIV-1DNA vaccine and rVV alone or combined.Immune response were detected with IFN-γ ELISPOT and ELISA.Results Specific IFN-γ cellular immune responese are obviously increased from 112 to 258(SFC/106SMNC) after modification in DNA vector,the antibody titer are enhanced from 101.7 to 102.3.There are no significant differences in rVV after gene optimization (600SFC/106SMNC,102.2).The combined immunization of DNA priming and rVV boost could elicit much higher immune responses than DNA or rVV alone immunization (788 to 2500 ) (SFC/106SMNC).Conclusion Gene modification is an important means for improving the expression level and immune effect,especially in the DNA vector.
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