人乳头瘤病毒6/11型L2片段的高效表达及免疫效果评价  

Immunogenicity of HPV6/11 Bivalent L2 Fusion Proteins Expressed in E.coli

作　　者：任皎[1] 张忠献[1,2] 赵莉[1] 郝明强[1] 田厚文[1] 谭文杰[1] 阮力[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]郑州大学中英分子肿瘤学研究中心,河南郑州450052

出　　处：《生物技术通讯》2016年第1期12-16,共5页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2006AA02Z421)

摘　　要：目的：原核表达人乳头瘤病毒（HPV）6型和11型L2 N端融合蛋白,并初步评价其免疫效果。方法：用重叠PCR将HPV6和HPV11次要衣壳蛋白L2基因5＇端片段融合,并在大肠杆菌中表达融合蛋白,纯化后与Al（OH）3佐剂配伍肌肉注射免疫BALB/c小鼠,用ELISA检测血清抗体,以基于假病毒的体外中和试验评价中和抗体水平。结果：ELISA结果显示,3种融合蛋白均能产生针对同型别L2蛋白的高滴度特异性Ig G抗体,滴度为1∶10 000～1∶200 000;体外中和试验显示,3种融合蛋白均能诱发中和抗体和交叉中和抗体,针对同型别的滴度最高可达1∶3200,对于高危型能产生一定水平的交叉中和抗体,滴度为1∶50～1∶800。结论：HPV6/11 L2融合蛋白能够诱发较强的体液免疫反应,产生较高的中和抗体及交叉中和抗体,为HPV L2新型预防性疫苗的研究提供了初步的实验基础。Objective： To assess the immunological efficacy of human papillomavirus（HPV） 6 and 11 bivalent L2 fusion proteins in immunized mice. Methods： We constructed bivalent fusion genes of HPV6/11 L2 5＇ termi- nus region by overlap PCR. And then, the L2 fusion genes were cloned into the pET9a vector. The proteins were expressed in E.coli. BALB/c mice were immunized with three purified L2 fusion proteins in combination with adju- vant AI（OH）3. The humoral immune response was measured by ELISA and in vitro HPV neutralizing expriment with pseudovirus. Results： The three HPV6/ll bivalent L2 fusion proteins could induce high specific serum lgG antibody titer, which was between 1：10 000 to 1：200 000; and also induce the neutralizing antibody titer against homogeneous and heterologous HPVs tested. The neutralizing antibody titer against homogeneous HPV was between 1：400 to 1：3200. The cross-neutralizing antibody titer against heterologous high risk HPVs pseudovirus was be- tween 1：50 to 1：800. Conclusion： HPV6/11 bivalent L2 fusion protein could induce specific high lgG antibodies and neutralizing and cross-neutralizing antibodies against HPV pseudovirus. This provides valuable experimental da- ta for in-depth study of a low-cost and broad spectrum of HPV L2 prophylactic vaccine.

关 键 词：人乳头瘤病毒 融合蛋白 预防性疫苗 中和抗体 次要衣壳蛋白L2 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]