柯萨奇B_2病毒VP_1基因疫苗诱导细胞免疫的初步研究  

Preliminary study on cell-mediated immunity induced by VP_1 gene vaccine against coxsackievirus B_2.

作　　者：郭宏[1] 田野[1] 曲秀芬[1] 刘胜旺 吴晓羽[1] 赵铁强[1] 李呼伦[3] 钟照华[3] 童光志 孟繁超[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院,150001 [2]中国农业科学院兽医生物技术国家重点实验室,150027 [3]哈尔滨医科大学基础医学院,150086

出　　处：《中国实用内科杂志》2003年第11期652-654,共3页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金资助 ( 3 0 2 70 5 5 3 )

摘　　要：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 a novel VP 1 gene vaccine against coxsackievirus B 2 and to evaluate the effect of the cell-mediated immunity induced by it.Methods The immunodominant capsid protein VP 1 gene of CVB 2 was amplified by reverse transcript polymerase chain reaction (RT-PCR) and pcDNA 3-CVB 2VP 1was constructed by molecular cloning.The cytotoxic T lymphocyte (CTL) activity was measured by standard 51Cr-release cytotoxicity assay eight weeks after BALB/c mice were immuned by pcDNA 3-CVB 2VP 1.Results The eukaryotic expression vector was pcDNA 3 and subcloning fragment was CVB 2VP 1.The CTL activity of pcDNA 3-CVB 2VP 1 group was higher than that of the control (P<0.01),with the highest percentage being (31.2±6.8)% when the ratio of effective cell to target cell was 50/1 (P<0.05).Conclusion Cell-mediated immunity can be induced by the constructed pcDNA 3-CVB 2VP 1 gene vaccine against coxsackievirus B 2.

关 键 词：柯萨奇B2病毒 VP1基因疫苗 诱导 细胞免疫 

分 类 号：R512.5[医药卫生—内科学]