Abstract
H7N9 viral infections pose a great threat to both animal and human health. This avian virus cannot be handled in level 2 biocontainment laboratories, substantially hindering evaluation of prophylactic vaccines and therapeutic agents. Here, we report a high-titer pseudoviral system with a bioluminescent reporter gene, enabling us to visually and quantitatively conduct analyses of virus replications in both tissue cultures and animals. For evaluation of immunogenicity of H7N9 vaccines, we developed an in vitro assay for neutralizing antibody measurement based on the pseudoviral system; results generated by the in vitro assay were found to be strongly correlated with those by either hemagglutination inhibition (HI) or micro-neutralization (MN) assay. Furthermore, we injected the viruses into Balb/c mice and observed dynamic distributions of the viruses in the animals, which provides an ideal imaging model for quantitative analyses of prophylactic and therapeutic monoclonal antibodies. Taken together, the pseudoviral systems reported here could be of great value for both in vitro and in vivo evaluations of vaccines and antiviral agents without the need of wild type H7N9 virus.
Highlights
H7N9 has caused annual human infections with high fatality rate since it was first identified in humans in 20131
Similar to clinical evaluation of seasonal influenza vaccines, in vitro assays such as hemagglutination inhibition (HI) and microneutralization (MN) assays are used to determine antibody titers in humans immunized with H7N9 vaccines; in preclinical studies, protections afforded by vaccines or antiviral agents were analyzed through monitoring survival rates or weight loss of the animals following challenges using wild type H7N9 viruses
Correspondence and requests for materials should be addressed to Y.W. www.nature.com/scientificreports/. To investigate whether this modified plasmid could be employed to improve the yield for H7N9 pseudovirus, we compared pSG3.Δenv-FlucΔnef with pNL4-3-Luc.R.E
Summary
H7N9 has caused annual human infections with high fatality rate since it was first identified in humans in 20131. Similar to clinical evaluation of seasonal influenza vaccines, in vitro assays such as hemagglutination inhibition (HI) and microneutralization (MN) assays are used to determine antibody titers in humans immunized with H7N9 vaccines; in preclinical studies, protections afforded by vaccines or antiviral agents were analyzed through monitoring survival rates or weight loss of the animals following challenges using wild type H7N9 viruses (wt H7N9). Both in vitro assays and in vivo animal studies require the use of live H7N9 viruses, which could hinder research and development of vaccines and antivirals. We succeeded in developing high-titer H7N9 pseudovirus and used them for in vitro and in vivo evaluation of vaccines and antibodies
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