HIV gag基因修饰在不同载体系统中对免疫效果的影响  

Effect of HIV gag Gene Modification on Immune Response in Dif ferent System

作　　者：高瑛瑛[1] 齐香荣[1] 黄保英[1] 王文玲[1] 邓瑶[1] 孟昕[1] 谭文杰[1] 阮力[1] 

机构地区：[1]中国疾病预防控制中心、病毒病预防控制所,北京102206

出　　处：《生物技术通讯》2012年第3期313-317,328,共6页Letters in Biotechnology

基　　金：中国综合性艾滋病研究(CIPRA)项目(1U19 A15191501);国家高技术研究发展计划(2003AA219080);国家"十一五"重大专项(2008ZX10001-012)

摘　　要：目的：了解gag基因修饰前后在不同载体系统中的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Gag靶抗原奠定实验基础。方法：将含有优化前后gag基因的HIV-1 DNA（pVRC）疫苗和重组痘苗病毒（rVV）载体疫苗单独或联合免疫BALB/c小鼠,利用IFN-γ酶联免疫斑点法和胞内细胞因子染色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Gag诱发的免疫效果。结果：DNA疫苗中gag基因修饰后细胞免疫反应由472提高至925 SFC/10^6MNC,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104.2提高至105.3,三针后则没有差别;而以rVV为载体的疫苗基因修饰前后细胞免疫反应（-320 SFC/10^6MNC）和抗体水平（-10^4.4）均没有差异。2种疫苗联合免疫均可显著提高Gag修饰前后在小鼠体内的免疫效果,基因修饰后细胞免疫反应由1700提高至2100 SFC/10^6MNC,抗体水平则没有差别。结论：gag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rVV疫苗单独免疫效果无明显影响。Objective： To understand the immune effect in different vectors after gag gene modification, study the Gag expression level differences on the immune response, to confirm the optimized gag 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-1 DNA vaccine and recombinant vaccinia virus（rVV） alone or combined. Immune responses were detected with IFN-γ ELISPOT and ELISA. Results： Specific IFN-γ cellular immune responses were obviously increased from 472 to 925 SFC/10^6 MNC） after modification in DNA vector, the antibody titer was enhanced from 104.2 to lOs3. There were no significant differences in rVV after gene optimization（320 SFC/10^6 MNC, 10^4.4）. The combined immunization of DNA priming and rVV boost could elicit much higher immune responses than DNA or rVV alone immunization（1700 to 2100 SFC/106 MNC）. Conclusion： Gene modification is an important means for improving the expression level and immune effect, especially in the DNA vector.

关 键 词：HIV-1 DNA疫苗 痘苗病毒载体 密码子优化 免疫反应 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]