pcDNA3/HPV16 E6真核表达质粒的构建及裸DNA注射动物实验观察

The construction of HPV16 E6 eukaryotic expression plasmid pcDNA3/E6 and its immunogenecity evaluation on animals

目的探讨ＨＰＶ１６Ｅ６基因ＤＮＡ诱发体液和细胞免疫反应的能力。方法利用基因工程技术构建了ＨＰＶ１６Ｅ６真核表达质粒ｐｃＤＮＡ３／Ｅ６，脂质体法转染Ｃｏｓ７细胞，肌肉注射免疫ＢＡＬＢ／ｃ小鼠，免疫组化技术检测抗体产生及抗原表达。结果被转染的Ｃｏｓ７细胞表达ＨＰＶ１６Ｅ６蛋白免疫小鼠产生抗ＨＰＶ１６Ｅ６抗体。结论这一结果为ＨＰＶ１６相关宫颈癌治疗性ＤＮＡ疫苗的研制提供了资料。

Objective To explore the ability of HPV16 E6 DNA to elicit humoral and cellular immunoresponse. Methods Eukaryotic expression plasmid pcDNA3/E6 was constructed with engineering technique,Cos7 cells were transfected,and BALB/c mice were immunized with the recombinant.Immunohistochemistry was used to identify the antigen expression and antibody production. Results The transfected Cos7 cells E6 protein and the immunized animals produced antibodies specifically react with caski cells harboring HPV16E6 gene. Conclusion The present results indicate that HPV16 E6 naked DNA could be used as a therapeutic vaccine for the therapy of HPV16 related cancers.