人乳头瘤病毒16型修饰后E6E7基因免疫原性增强  

Modifid E6E7 gene of HPV increases immunogenicity of human papillomavirus 16

作　　者：许雪梅[1] 朱明昭[1] 张明策[1] 刘晓娟[1] 宋国兴[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所生物物理室,北京100005

出　　处：《基础医学与临床》2003年第3期269-273,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金 (3970 0 131) ;国家"十五"86 3课题 (2 0 0 1AA2 15 2 2 1)

摘　　要：利用突变修饰后消除转化活性并保留抗原性的中国山东地方株人乳头瘤病毒 16型 (humanpapillomavirustype16 ,HPV16 )E6E7融合基因 (fmE6E7) ,研制治疗HPV16相关疾病的DNA疫苗。用PCR扩增fmE6E7基因后 ,插入真核表达质粒获得pVR10 12 fmE6E7,瞬时转染Cos 7细胞 ,免疫荧光法检测证实其表达后 ,在C5 7BL 6小鼠后腿肌肉进行裸DNA免疫 ,5 1Cr释放法体外分析免疫鼠的细胞毒性T淋巴细胞活性 (cytotoxicTlymphocyte,CTL) ,间接ELISA法检测免疫鼠血清中E7特异性抗体。研究表明修饰后的中国地方株E6E7融合基因可诱导机体产生特异的抗体反应和CTL反应 ,与单独野生型E7基因免疫相比 ,E6E7融和基因可更好的活化CTL反应。表明修饰后消除转化活性的中国地方株E6E7融合基因可作为HPV16治疗性DNA疫苗的靶基因。To develop a therapeutic vaccine against HPV16 with the modified HPV16E6E7 fused gene (fmE6E7) from a cervical cancer patient,that has the antigenicity of E6 and E7 but not transformational activity. The modified HPV16E6E7gene was amplified by PCR and then inserted into eukaryotic expression plasmid for constructing pVR1012 fmE6E7. After confirmed the ability to express in eukaryotic cells in vitro, the pVR1012 fmE6E7 plasmid was inoculated directly into the muscles on the posterior leg of C57BL/6 mice, the activities of cytotoxic T lymphocytes (CTL) isolated from immunized mice were tested in vitro with 51 Cr specific release assay and the specific antibody in sera was analyzed by ELISA. The CTLs and E7 specific antibody from immunized mice were efficaciously activated by pVR1012 fmE6E7 plasmid; higher activity of CTL were be produced by pVR1012 fmE6E7 than pVR1012 E7. The modified E6E7 gene has been proved to be a candidate of DNA vaccine.

关 键 词：人乳头瘤病毒16型 基因修饰 E6E7基因 免疫原性增强 DNA疫苗 宫颈癌 癌前病变 

分 类 号：R392[医药卫生—免疫学]