＜正＞ Background Although DNA vaccine is considered as the next generation of vaccine,most DNA vaccine candidates arestill suffering from the relatively weak immunogenicity despite the increased dosage of plasmid DNA administered.Inorder to enhance the immune responses elicited by a codon-optimized HIV gag DNA vaccine,a modified plasmid vectorpDRVI1.0 and a booster immunization with replicating Tiantan vaccinia（RTV）strain expressing the same gene wereemployed.Methods Vector pDRVI1.0 was constructed through inserting the 72-bp element from the SV40 enhancer,which wasreported promoting nuclear transport of plasmid DNA,to the upstream of cytomegalovirus enhancer/promoter region ofthe plasmid vector pVR1012.Gene expression levels from expression plasmids based on pDRVI1.0 and pVR1012 weretested.Humoral and cellular immune responses induced by DNA vaccine alone or DNA prime-RTV boost regimen weredetermined in mice.Results It was shown that the 72-bp element significantly enhanced the gene expression level in non-dividing cells.gag-specific humoral and cellular immune responses induced by DNA vaccination were both significantly improved,whilethe Th1/Th2 balance was not obviously affected by the 72-bp element.RTV boosting further significantly enhancedDNA vaccine-primed antibody and T cell responses in a Th1-biased manner.Conclusions The 72-bp SV40 enhancer element should be included in the DNA vaccine vector and RTV strain is avery efficient live vector for boosting immunization.
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