Construction and immunogenicity evaluation of human papillomavirus type 16 epitope chimeric virus-like particles
GUO Jian1, ZHANG Li2, CHENG Li-yuan1, LI Yu-hua2, YANG Hao-jie1, TIAN Yi-fan1, ZHU Jie3, LIU Wei1
1.Jilin Medical University, Changchun 132012, China; 2. Jilin Central Hospital, Changchun 132111, China; 3. College of Life Sciences, Jilin University, Changchun 130012,China
Abstract:To prepare E749-57 epitope chimeric VLPs(cVLPs-E749-57)and analyze its immunogenicity, the E749-57 epitope was inserted into the optimal chimeric site on HPV16 L1 which was predicted by the molecular simulation software of Discovery Studio. Then the recombinant plasmid pET28a-16L1-E749-57 was constructed, the HPV16L1-E749-57 protein was induced to express in E. coli and purified by affinity chromatography using a nickel column. After in-vitro recombination, cVLPs-E749-57 was confirmed by dynamic light scattering and electron microscopy. Mice were immunized with cVLPs-E749-57, and the neutralizing antibody titer of immune serum was detected by a pseudoviral neutralization test. The levels of Th1 and Th2 cytokines were detected by flow multifactor technology. The results showed that the optimal chimeric site was HI loop (355/356), HPV16L1-E749-57 protein was expressed and cVLPs-E749-57 was assembled successfully. After 3 times of immunizing, the average log10 of serum neutralizing antibody titer reached 4.23, which was slightly lower than the wild-type VLPs (the average log10 was 4.45). However, compared with wild-type VLPs, cVLPs-E749-57 could induce significantly higher levels of Th1-type cytokines (INF-γ, IL-2, TNF-α). In conclusion, the cVLPs-E749-57 could stimulate the body to produce strong humoral and cellular immune response simultaneously, which might have both preventive and therapeutic effects. It lays the foundation for the development of therapeutic vaccines against cervical cancer.
郭健, 张力, 陈力源, 李玉华, 杨皓杰, 田一凡, 朱洁, 刘微. 人乳头瘤病毒16型表位嵌合病毒样颗粒的构建及免疫原性研究[J]. 中国人兽共患病学报, 2020, 36(9): 689-695.
GUO Jian, ZHANG Li, CHENG Li-yuan, LI Yu-hua, YANG Hao-jie, TIAN Yi-fan, ZHU Jie, LIU Wei. Construction and immunogenicity evaluation of human papillomavirus type 16 epitope chimeric virus-like particles. Chinese Journal of Zoonoses, 2020, 36(9): 689-695.
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