人乳头瘤病毒16型E7DNA疫苗的构建及其诱导特异性淋巴细胞增殖的研究

目的 :探讨人乳头瘤病毒 16型 (HPV16 )E7DNA疫苗诱导机体特异性细胞免疫应答的情况。方法 :采用分子克隆技术 ,构建HPV16野生型E7基因的真核表达重组体 ,将其转化大肠杆菌JM10 9进行筛选 ,通过限制性内切酶酶切鉴定和PCR分析 ,证明重组质粒中目的基因插入片段及载体DNA大小、方向、插入位点均正确 ,获得HPV16E7DNA疫苗 ,将E7DNA疫苗经皮内注射免疫动物 ,MTT比色法体外检测特异性淋巴细胞增殖反应。结果 :野生型E7DNA疫苗免疫组脾淋巴细胞在体外受到E7蛋白的再次刺激后出现特异性淋巴细胞增殖反应。结论 :野生型E7DNA疫苗可诱导特异性的细胞免疫应答 ,皮内注射HPVDNA疫苗是一种简便有效的免疫接种途径

Construction of HPV16 E7 DNA Vaccine and Its Induction of Specific Lymphoproliferation

Objective: To study the specific cell mediated immune response which was induced by HPV16 E7 DNA vaccine. Methods: We cloned ysE7 gene into the eukaryotic expression vector pcDNA3.1 . Then the recombinant was transformed E.coli JM109, screening and identified by PCR and restriction analysis. It was proved that the length, inserted locatin and direction of the target gene which was inserted into the recombinant was correct. Finally, we obtained HPV16 ysE7 DNA vaccine named pcDNA3.1 (ysE7). The HPV16 ysE7 DNA vaccine was injected by intradermal administration. The specific lymphoproliferation after E7 protein restimulation in vitro was detected by MTT colorimetric assay. Results: The ysE7 DNA vaccine administration group had specific lymphoproliferation after restimulating by E7 protein. Conclusion: The ysE7 DNA vaccine can elicit specific cell mediated immune response. The intradermal administration is a simple and efficient way for application HPV DNA vaccine.