Immunogenicity and receptor binding ability of the virus-like particle of the GII.3P12 human norovirus
10.3760/cma.j.cn112866-20210922-00168-1
- VernacularTitle:人源诺如病毒GII.3[P12]型病毒样颗粒的免疫原性及受体结合能力
- Author:
Linping WANG
1
;
Junshan GAO
;
Liang XUE
;
Dapeng WANG
;
Yanhui LIANG
;
Xiaojing HONG
;
Jumei ZHANG
;
Qingping WU
Author Information
1. 喀什大学生命与地理科学学院 新疆帕米尔高原生物资源与生态重点实验室，喀什 844000
- Keywords:
Human norovirus;
GII.3[P12];
Virus-like particle;
Polyclonal antibody;
Receptor binding function;
Immunogenicity
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(5):514-520
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare the virus-like particle (VLP) of the GII.3[P12] human norovirus (HuNoV) strain GZ2013-L20 in Guangzhou and its polyclonal antibody, and systematically characterize its immunogenicity and receptor binding ability, which would provide data for prevention and control of HuNoV.Methods:ORF2 gene was amplified from the genome of the GZ2013-L20 strain to construct the recombinant transposon vector, which was further transformed into Escherichia coli DH10 Bac to develop the recombinant baculovirus Bacmid-L20-ORF2. VLP was expressed in the sf9 insect cells and then purified. Transmission electron microscopy, SDS-PAGE, Western blot (WB), and receptor binding experiments were performed to characterize the purified VLP. In addition, the polyclonal antibody from the immunized mice was evaluated by indirect enzyme-linked immunosorbent assay (ELISA) and the blocking test of receptor binding. Results:The recombinant baculovirus plasmid Bacmid-L20-ORF2 was constructed, and the target VLP was successfully obtained. The result by the transmission electron microscope demonstrated that the VLP were about 30 nm in diameter. SDS-PAGE and WB analyses showed that the protein’s relative molecular mass (Mr. ×10 3) was about 58. The result of receptor binding experiments showed that the VLP could bind to the secretory salivary receptors (types of A, B, AB and O), non-secretory salivary receptors (O type) and the porcine gastric mucin. The polyclonal antibody with a titer of 2 × 10 5 was detected in the immunized mice, which showed strong cross-immunoreactivity with capsid proteins of 20 (20/28) HuNoV genotypes. In addition, the result of blocking tests of receptor binding showed that the VLP polyclonal antibody only blocked the viral VLP of the same genotype, but had no neutralizing effects on the VLPs of GII.2, GII.4, GII.8 and GII.17. Conclusions:The VLP of GII.3[P12] HuNoV Guangzhou strain showed strong binding ability to both secretory and non-secretory salivary receptors, and its polyclonal antibody showed a broad spectrum of immunobinding, but its neutralization blocking feature was effective only against the virus of the same genotype. The result provide basic data for rational design of vaccine development.
