共表达HPV16型E6和E7突变基因重组痘苗病毒的构建及其抗肿瘤免疫效果的观察  

Construction of recombinant vaccinia virus co-expressing mutant E6 plus E7 proteins and detection of its immunogenicity and antitumor response

作　　者：职慧军[1] 韩立群[1] 任皎[1] 田厚文[1] 骆卫锋[1] 梁雨[1] 阮力[1] 

机构地区：[1]中国预防医学科学院病毒学研究所遗传免疫研究室,北京100052

出　　处：《中华实验和临床病毒学杂志》2002年第4期341-344,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家"九五"科技攻关重点项目基金资助 (96 90 6 0 1 11)

摘　　要：目的　构建用于子宫颈癌治疗的HPV16型E6和E7重组痘苗病毒实验性疫苗株 ,并对其抗肿瘤免疫效果进行初步评价。方法　以痘苗病毒为载体、利用同源重组技术构建共表达HPV16E6和E7基因的重组痘苗病毒。该病毒免疫C57BL 6小鼠后 ,检测其免疫原性和抗移植瘤生长情况。结果　PCR结果显示 ,重组病毒VmE6E7的TK基因内插入了分别由痘苗病毒早晚期启动子H6和7.5K表达的ME6和ME7 1基因。动物实验结果表明 ,rVmE6E7在C57BL 6小鼠体内可诱发E6和E7特异性抗体产生 ,被免疫小鼠能够抵抗HPV16E6E7转化的同系肿瘤细胞的攻击。结论　获得 1株用于宫颈癌治疗的HPV16型实验疫苗株 ,为进一步研制人用HPV16型疫苗株奠定了基础。Objective To generate a candidate HPV16 vaccine for experimental and the rapeutical use for cervical cancer. Methods The mutants of HPV16 early E6 and E7 genes were inserted into a vaccinia virus expression vector. A strain of recombinant vaccinia virus was constructed through homologous recombination. Results PCR showed that the mutant E6 and E7 genes were located at TK gene region of vaccinia virus Tiantan strain in a head to head orientation under the control of early/late promoters, H6 and 7.5K respectively. Studies in mice indicated that VmE6E7 could elicit specific antibodies against E6 and E7, and retarded or prevented tumor development in a proportion of C 57 BL/6 mice challenged by syngeneic HPV16E6 and E7 transformed tumor cells. Conclusion The success in constructing VmE6E7 provides a basis for the further development of HPV16 therapeutic vaccine.

关 键 词：抗肿瘤免疫效果 16型人乳头状瘤病毒 宫颈肿瘤 DNA疫苗 E6蛋白 E7蛋白 治疗性疫苗实验株 

分 类 号：R392[医药卫生—免疫学]