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Viruses 2016, 8(2), 49; doi:10.3390/v8020049

In Vitro and in Vivo Evaluation of Mutations in the NS Region of Lineage 2 West Nile Virus Associated with Neuroinvasiveness in a Mammalian Model

Veterinary Diagnostic Directorate, National Food Chain Safety Office, H-1143, Budapest, Hungary
Viroscience Laboratory, Erasmus Medical Centre, 3015CN, Rotterdam, The Netherlands
Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, H-1143, Budapest, Hungary
Department of Microbiology and Infectious Diseases, Faculty of Veterinary Science, Szent István University, H-1143, Budapest, Hungary
Viral Zoonoses, Emerging and Vector-Borne Infections Group, Institute of Virology, University of Veterinary Medicine, A-1210, Vienna, Austria
K.S.-G. and S. L. contributed equally to this work.
Author to whom correspondence should be addressed.
Academic Editor: Andrew Mehle
Received: 30 September 2015 / Revised: 19 January 2016 / Accepted: 9 February 2016 / Published: 19 February 2016
(This article belongs to the Section Animal Viruses)
View Full-Text   |   Download PDF [2277 KB, uploaded 19 February 2016]   |  


West Nile virus (WNV) strains may differ significantly in neuroinvasiveness in vertebrate hosts. In contrast to genetic lineage 1 WNVs, molecular determinants of pathogenic lineage 2 strains have not been experimentally confirmed so far. A full-length infectious clone of a neurovirulent WNV lineage 2 strain (578/10; Central Europe) was generated and amino acid substitutions that have been shown to attenuate lineage 1 WNVs were introduced into the nonstructural proteins (NS1 (P250L), NS2A (A30P), NS3 (P249H) NS4B (P38G, C102S, E249G)). The mouse neuroinvasive phenotype of each mutant virus was examined following intraperitoneal inoculation of C57BL/6 mice. Only the NS1-P250L mutation was associated with a significant attenuation of virulence in mice compared to the wild-type. Multiplication kinetics in cell culture revealed significantly lower infectious virus titres for the NS1 mutant compared to the wild-type, as well as significantly lower amounts of positive and negative stranded RNA. View Full-Text
Keywords: West Nile virus; lineage 2; virulence markers; infectious clone West Nile virus; lineage 2; virulence markers; infectious clone

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Szentpáli-Gavallér, K.; Lim, S.M.; Dencső, L.; Bányai, K.; Koraka, P.; Osterhaus, A.D.; Martina, B.E.; Bakonyi, T.; Bálint, Á. In Vitro and in Vivo Evaluation of Mutations in the NS Region of Lineage 2 West Nile Virus Associated with Neuroinvasiveness in a Mammalian Model. Viruses 2016, 8, 49.

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