基于汉逊酵母 HPV68b L1蛋白的 VLPs免疫学效果研究  

Immunogenicity study of human papillomavirus type 68b virus-like particles expressed in a Hansenu-la polymorpha strain

作　　者：陈实[1] 靳玉琴[1] 杜丽芳[1] 梁宇[1] 郭舒杨 李启明[1] 

机构地区：[1]北京生物制品研究所有限责任公司第六研究室,101111

出　　处：《中华微生物学和免疫学杂志》2015年第9期653-656,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家科技部重大新药创制（2013zX09402302）;北京市科委统筹项目（Z131100006513007）

摘　　要：目的：对重组表达HPV68 b L1蛋白的汉逊酵母工程菌进行高密度发酵，并对L1蛋白形成的类病毒颗粒（virus-like particles，VLPs）进行纯化、鉴定及免疫学效果评价。方法 HPV68b L1的酵母工程菌经过发酵，系列纯化和颗粒重构后获得VLPs抗原，对该抗原进行蛋白纯度、颗粒形态及免疫原性等检定，与铝佐剂吸附后免疫动物，利用假病毒体外中和试验评价其对HPV68 b型及HPV68a型的保护效果。结果发酵后的工程菌经过纯化和颗粒重构后，获得纯度大于95％的VLPs，电镜结果显示，其颗粒形态均一，大小与天然病毒颗粒接近。假病毒体外中和试验结果显示， HPV68b型VLPs抗原可诱导小鼠体内产生较高滴度的中和抗体，同时对HPV68a型也具有一定的交叉保护作用。结论验证了重组HPV68 b型VLPs蛋白用于疫苗生产的可行性，可作为多价HPV疫苗的组分。Objective To evaluate the immunogenicity of the recombinant human papillomavirus type 68b （HPV68b） virus-like particles（VLPs）in a mouse model.Methods The L1 protein of HPV type 68b was successful expressed in the Hansenula polymorpha strain （NVSI-68b-1）.Processes including purifi-cation and reconstitution were performed to achieve pure HPV 68b VLPs.The purity, morphology and immu-nogenicity of the purified HPV 68 b VLPs were further analyzed .The BALB/c mice were immunized with HPV68b VLPs formulated on aluminum adjuvant .Pseudovirus-neutralizing antibody （ PsV NAb） assay was performed to detect the neutralizing antibodies in serum samples .Results The HPV 68 b L1 VLPs were ob-tained as indicated by the results of SDS-PAGE, Western blot assay , HPLC, electron microscopy and dy-namic light scattering with a high purity of 95%.Transmission electron microscopy and dynamic light scat-tering analysis revealed that the HPV68b L1 VLPs resembled the native virus with an average particle diame-ter of 50 nm.High levels of HPV68b-neutralizing antibodies were detected in serum samples from the mice immunized with HPV68b L1 VLPs.Moreover, a cross-protective efficacy of HPV68b L1 VLPs for HPV68a was observed .Conclusion This study suggested that the recombinant HPV 68 b VLPs expressed in a Han-senula polymorpha strain might be used as a potential candidate for the development of HPV vaccine .

关 键 词：汉逊酵母 HPV68 交叉保护 

分 类 号：R392[医药卫生—免疫学]