Journal
VIROLOGY
Volume 346, Issue 1, Pages 53-65Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2005.10.030
Keywords
West Nile viris; flavivirus; antibody-ruediated neutralization; reporter virus particle; pseudotype
Categories
Funding
- Intramural NIH HHS Funding Source: Medline
- NCRR NIH HHS [F31 RR05074] Funding Source: Medline
- NIAID NIH HHS [U01 AI061373, U54 AI57173] Funding Source: Medline
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West Nile virus (WNV) is a neurotropic flavivirus within the Japanese encephalitis antigenic complex that is responsible for causing West Nile encephalitis in humans. The surface of WNV virions is covered by a highly ordered icosahedral array of envelope proteins that is responsible for mediating attachment and fusion with target cells. These envelope proteins are also primary targets for the generation of neutralizing antibodies in vivo. In this study, we describe a novel approach for measuring antibody-mediated neutralization of WNV infection using Virus-like particles that measure infection as a function of reporter gene expression. These reporter virus particles (RVPs) are produced by complementation of a subgenomic replicon with WNV structural proteins provided in trans using conventional DNA expression vectors. The precision and accuracy of this approach stern from an ability to measure the Outcome of the interaction between antibody and viral antigens under conditions that satisfy the assumptions of the law of mass action as applied to virus neutralization. In addition to its quantitative strengths, this approach allows the production of WNV RVPs bearing the prM-E proteins of different WNV strains and mutants, offering considerable flexibility for the study of the humoral immune response to WNV in vitro. WNV RVPs are capable of only a Single round of infection, can be used under BSL-2 conditions, and offer a rapid and quantitative approach for detecting Virus entry and its inhibition by neutralizing antibody. Published by Elsevier Inc.
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