title: A rapid and quantitative assay for measuring antibody-mediated neutralization of West Nile virus infection.
abstract: 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 sub-genomic replicon with WNV structural proteins provided in trans using conventional DNA expression vectors. The precision and accuracy of this approach stem 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.
authors.affiliation: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA. piersontc@mail.nih.gov
authors.forename: Theodore C
authors.initials: TC
authors.lastname: Pierson
authors.forename: Melissa D
authors.initials: MD
authors.lastname: Sánchez
authors.forename: Bridget A
authors.initials: BA
authors.lastname: Puffer
authors.forename: Asim A
authors.initials: AA
authors.lastname: Ahmed
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Geiss
authors.forename: Laura E
authors.initials: LE
authors.lastname: Valentine
authors.forename: Louis A
authors.initials: LA
authors.lastname: Altamura
authors.forename: Michael S
authors.initials: MS
authors.lastname: Diamond
authors.forename: Robert W
authors.initials: RW
authors.lastname: Doms
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D009500:Neutralization Tests; D012093:Replicon; D013997:Time Factors; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology; D014771:Virion / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014901:West Nile Fever / Q000276:immunology; D014902:West Nile virus / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity*
chemical_list: D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus