title: Development of in vitro and in vivo rabies virus neutralization assays based on a high-titer pseudovirus system.
abstract: Pseudoviruses are useful virological tools because of their safety and versatility; however the low titer of these viruses substantially limits their wider applications. We developed a highly efficient pseudovirus production system capable of yielding 100 times more rabies pseudovirus than the traditional method. Employing the high-titer pseudoviruses, we have developed robust in vitro and in vivo neutralization assays for the evaluation of rabies vaccine, which traditionally relies on live-virus based assays. Compared with current rapid fluorescent focus inhibition test (RFFIT), our in vitro pseudovirus-based neutralization assay (PBNA) is much less labor-intensive while demonstrating better reproducibility. Moreover, the in vivo PBNA assay was also found to be superior to the live virus based assay. Following intravenous administration, the pseudovirus effectively infected the mice, with dynamic viral distributions being sequentially observed in spleen, liver and brain. Furthermore, data from in vivo PBNA showed great agreement with those generated from the live virus model but with the experimental time significantly reduced from 2 weeks to 3 days. Taken together, the effective pseudovirus production system facilitated the development of novel PBNA assays which could replace live virus-based traditional assays due to its safety, rapidity, reproducibility and high throughput capacity.
authors.affiliation: Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Nie
authors.affiliation: Division of Arboviral Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Ma
authors.affiliation: Division of Arboviral Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Shouchun
authors.initials: S
authors.lastname: Cao
authors.affiliation: Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Weijin
authors.initials: W
authors.lastname: Huang
authors.affiliation: Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, On K1A 0K9, Canada.|Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, On, Canada.
authors.forename: Xuguang
authors.initials: X
authors.lastname: Li
authors.affiliation: Division of Arboviral Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Li
authors.affiliation: Division of HIV/AIDS and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantanxili, Beijing 100050, China.
authors.forename: Youchun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D009500:Neutralization Tests / Q000379:methods*; D011819:Rabies Vaccines / Q000276:immunology*; D011820:Rabies virus / Q000276:immunology* / Q000502:physiology; D015203:Reproducibility of Results; D019562:Viral Load
chemical_list: D011819:Rabies Vaccines