论文部分内容阅读
为探索Semliki森林病毒 (SFV)衍生的复制型DNA载体可否用于HIV疫苗的候选载体 ,对该载体与传统DNA疫苗载体对HIV - 1Pr5 5 gag的表达与体液免疫原性进行了系统比较研究。将野生型 (wtgag)及密码子改造(syngag)的HIV - 1ⅢBgag 基因分别克隆于SFVDNA载体及传统DNA疫苗载体 [pCDNA3 1(+) ],对其Pr5 5 gag细胞内表达水平、Pr5 5 gag病毒样颗粒释放、以及在BALB/c鼠的体液免疫原性进行了比较。在 2 93T、H12 99、C2C12和BHK细胞系中 ,SFV -wtgag可以Rev非依赖方式有效表达Pr5 5 gag ,而 pC -wtgag转染的细胞不能有效表达Pr5 5 gag,从而不能诱导小鼠产生免疫反应。虽然SFV质粒的细胞转化效率明显低于pCDNA载体 ,SFV -wtgag和SFV -syngag在细胞内Pr5 5 gag的表达量与 pC -syngag相似 ,而Pr5 5 gag病毒样颗粒的释放明显低于 pC -syngag。在肌内注射免疫的小鼠中 ,低剂量 (0 1和 1 0 μg)的SFV及 pCDNAgag表达质粒均未诱导出GAG特异性免疫反应。在高剂量 (10 ,30 ,10 0 μg)免疫组中 ,与SFVgag表达质粒相比 ,pC -syngag可诱导出较高水平的TH1型GAG特异性抗体。SFV -syngag较SFV -wtgag可诱导出高水平的体液免疫反应。结果提示 ,SFV衍生的复制子单独使用不能在小鼠诱导出优于传统DNA疫苗载体的HIV - 1GAG特异性?
In order to explore whether Semliki forest virus (SFV) - derived replicative DNA vector can be used as a candidate vector for HIV vaccine, we systematically compared the expression of this vector with that of the conventional DNA vaccine against HIV - 1Pr5 5 gag and humoral immunogenicity. The wild-type (wtgag) and codon-modified HIV-1ⅢBgag genes were cloned into the SFVDNA vector and the conventional DNA vaccine vector [pCDNA3 1 (+)], respectively. The expression levels of Pr5 5 gag, Like particle release, as well as humoral immunogenicity in BALB / c mice. SFV-wtgag efficiently expressed Pr5 5 gag in 2 93T, H12 99, C2C12 and BHK cell lines in a Rev-independent manner, whereas pC-wtgag-transfected cells did not efficiently express Pr5 5 gag and thus did not induce immunization in mice reaction. Although the cell transformation efficiency of SFV plasmid was significantly lower than that of pCDNA vector, expression of Pr5 5 gag was similar to that of pC -syngag in SFV-wtgag and SFV-syngag cells, while the release of Pr5 5 gag virus-like particles was significantly lower than that of pC -syngag . Low-dose (0 1 and 10 μg) SFV and pCDNAgag expression plasmids did not induce GAG-specific immune responses in mice immunized intramuscularly. In the high-dose (10, 30, 100 μg) immunized group, pC-syngag induced higher levels of TH1-type GAG-specific antibodies compared to the SFVgag expression plasmid. SFV-syngag induced a higher level of humoral immune response than SFV-wtgag. The results suggest that SFV derived replicon alone can not induce HIV - 1 GAG specificity in mice that is superior to conventional DNA vaccine vectors.