HIV-1B亚型gp120基因密码子优化前后免疫原性的比较

Study on the Immunogenicity of HIV-1 Clade B gp120 Gene before and after Codon Optimization

对HIV 1B亚型gp120基因按照哺乳动物优势密码子的使用原则进行优化,以Westernblot方法比较其体外表达量。将优化前的野生型gp120基因和改造后的modgp120基因插入重组腺伴随病毒载体,构建了重组病毒rAAV wtgp120和rAAV modgp120,比较两者免疫Balb/C小鼠后的抗体和CTL应答。Westernblot检测结果显示:优化后基因的体外表达量明显高于野生型基因,rAAV modgp120与rAAV wtgp120相比可更好地诱导Balb/C小鼠的CTL应答,但检测不到明显的抗体反应。由此得出结论,优化后gp120基因的体外表达量明显高于野生型基因,并且可以诱导更强的特异性CTL应答,但检测不到gp120抗体。

To compare the immunogenicity of HIV-1 clade B gp120 gene using optimized codons with wild-type gene based on recombinant adeno-associated virus vector(rAAV),we,firstly,generated a synthetic HIV-1 gp120 sequence in which most wide-type codons were replaced with codons from highly expressed human genes(modgp120) and identified its better expression in vitro than that of wild-type gene(wtgp120)using Western blot;secondly,we inserted wtgp120 and modgp120 into rAAV vector and produced pure rAAV-wtgp120 and rAAV-modgp120.Balb/C mice were immunized with rAAV-wtgp120 and rAAV-modgp120 separately,the antibody and CTL responses were tested.Results indicated that modgp120 gene expressed more antigen in vitro and induced higher specific CTL response in vivo than that of wild-type gp120 gene.There was no detectable antibody to gp120.