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目的 构建柯萨奇B2 病毒 (CVB2 )VP1侯选基因疫苗 ,评价其诱导细胞免疫的效果。方法 采用逆转录PCR技术扩增CVB2 的主要中和抗原VP1基因 ,通过分子克隆构建 pcDNA3 CVB2 VP1基因免疫质粒并免疫BALB/c小鼠 ,51Cr释放法测定CTL杀伤效应。结果 基因免疫质粒表达载体为 pcDNA3 ,亚克隆片段为CVB2 VP1,pcDNA3 CVB2 VP1接种组CTL特异性杀伤百分率均高于 pcDNA3 组 (P <0 0 1) ,E/T为 5 0∶1时特异性CTL杀伤百分率最高 ,为 (31 2± 6 8) % (P <0 0 5 )。结论 pcDNA3 -CVB2 VP1可诱导小鼠产生细胞免疫。
Objective To construct VP1 candidate gene vaccine of Coxsackie B2 virus (CVB2) and evaluate its effect on inducing cellular immunity. Methods VP1 gene of CVB2 was amplified by reverse transcription polymerase chain reaction (RT-PCR). The immunophenotype of pcDNA3 CVB2 VP1 gene was constructed by immunization with BALB / c mice and the CTL killing effect was determined by 51Cr release assay. Results The results of gene immunization with plasmid pcDNA3 and subclone were CVB2 VP1. The CTL specific killing rate of pcDNA3 CVB2 VP1 inoculated group was higher than that in pcDNA3 group (P <0.01), and the specificity of E / T was 5 0:1 The highest percentage of CTL killing was (31 2 ± 6 8)% (P <0 05). Conclusion pcDNA3-CVBV VP1 can induce cellular immunity in mice.