Immunogenicity and receptor binding ability of the virus-like particle of the GII.3[P12] human norovirus
Wang Linping, Gao Junshan, Xue Liang, Wang Dapeng, Liang Yanhui, Hong Xiaojing, Zhang Jumei, Wu Qingping
Abstract
ObjectiveTo 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.
MethodsORF2 gene was amplified from the genome of the GZ2013-L20 strain to construct the recombinant transposon vector, which was further transformed into Escherichia coli DH10Bac 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.
ResultsThe 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. ×103) 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 × 105 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.
ConclusionsThe 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.
Key words:
Human norovirus; GII.3[P12]; Virus-like particle; Polyclonal antibody; Receptor binding function; Immunogenicity
Contributor Information
Wang Linping
College of Life and Geographic Sciences, Kashi University, Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region, Kashi 844000, China
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Gao Junshan
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Xue Liang
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Wang Dapeng
Department of Food Science and Engineering, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China
Liang Yanhui
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Hong Xiaojing
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Zhang Jumei
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
Wu Qingping
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Key Laboratory of Agricultural Microbiomics and Precision Application, Ministry of Agriculture and Rural Affairs, Guangzhou 510070, China
