Document Detail


Microarray hybridization for assessment of the genetic stability of chimeric West Nile/dengue 4 virus.
MedLine Citation:
PMID:  21360544     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Genetic stability is an important characteristic of live viral vaccines because an accumulation of mutants can cause reversion to a virulent phenotype as well as a loss of immunogenic properties. This study was aimed at evaluating the genetic stability of a live attenuated West Nile (WN) virus vaccine candidate that was generated by replacing the pre-membrane and envelope protein genes of dengue 4 virus with those from WN. Chimeric virus was serially propagated in Vero, SH-SY5Y human neuroblastoma and HeLa cells and screened for point mutations using hybridization with microarrays of overlapping oligonucleotide probes covering the entire genome. The analysis revealed several spontaneous mutations that led to amino acid changes, most of which were located in the envelope (E) and non-structural NS4A, NS4B, and NS5 proteins. Viruses passaged in Vero and SH-SY5Y cells shared two common mutations: G(2337) C (Met(457) Ile) in the E gene and A(6751) G (Lys(125) Arg) in the NS4A gene. Quantitative assessment of the contents of these mutants in viral stocks indicated that they accumulated independently with different kinetics during propagation in cell cultures. Mutant viruses grew better in Vero cells compared to the parental virus, suggesting that they have a higher fitness. When tested in newborn mice, the cell culture-passaged viruses did not exhibit increased neurovirulence. The approach described in this article could be useful for monitoring the molecular consistency and quality control of vaccine strains.
Authors:
Majid Laassri; Bella Bidzhieva; James Speicher; Alexander G Pletnev; Konstantin Chumakov
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2011-02-25
Journal Detail:
Title:  Journal of medical virology     Volume:  83     ISSN:  1096-9071     ISO Abbreviation:  J. Med. Virol.     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-03-17     Completed Date:  2011-07-15     Revised Date:  2013-06-30    
Medline Journal Info:
Nlm Unique ID:  7705876     Medline TA:  J Med Virol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  910-20     Citation Subset:  IM    
Copyright Information:
Copyright © 2011 Wiley-Liss, Inc.
Affiliation:
Laboratory of Method Development, Center for Biologics Evaluation and Research, US Food and Drug Administration, Rockville, Maryland 20852-1448, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line
DNA Primers / genetics
Dengue Virus / genetics*
Genomic Instability*
Humans
Microarray Analysis / methods
Nucleic Acid Hybridization / methods
Point Mutation
Recombination, Genetic*
Serial Passage
Virology / methods
Virus Cultivation
West Nile Virus Vaccines / genetics*
West Nile virus / genetics*,  growth & development
Grant Support
ID/Acronym/Agency:
224-06-1322/Y1-AI-6153-01/AI/NIAID NIH HHS; ZIA AI000637-20/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/DNA Primers; 0/West Nile Virus Vaccines
Comments/Corrections

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