Establishment of an in vivo neutralization model based on H5N1 pseudovirus
10.3760/cma.j.cn112309-20230206-00023
- VernacularTitle:基于H5N1假病毒的体内中和试验模型的建立
- Author:
Nan CHEN
1
;
Xiangjun HAO
;
Jing WANG
;
Guojiang CHEN
;
Chunxia QIAO
;
Xinying LI
;
Beifen SHEN
;
Jiannan FENG
;
Lihui CHAI
;
He XIAO
Author Information
1. 军事科学院军事医学研究院毒物药物研究所，北京 100850
- Keywords:
Influenza virus;
Pseudovirus;
In vivo model;
Neutralization test
- From:
Chinese Journal of Microbiology and Immunology
2023;43(6):458-463
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.
