title: Characterization of neutralizing antibodies to West Nile virus.
abstract: We produced nine monoclonal antibodies (MAbs) directed against the West Nile virus E glycoprotein using three different immunization strategies: inactivated virus, naked DNA, and recombinant protein. Most of the MAbs bound to conformation dependent epitopes in domain III of the E protein. Four of the MAbs neutralized WNV infection and bound to the same region of domain III with high affinity. The neutralizing MAbs were obtained from mice immunized with inactivated virus alone or in combination with a DNA plasmid. In contrast, MAbs obtained by immunization with a soluble version of the E glycoprotein did not exhibit neutralizing activity. These non-neutralizing antibodies were cross-reactive with several other flaviviruses, including Saint Louis encephalitis, Japanese encephalitis, Yellow Fever and Powassan viruses. Interestingly, some non-neutralizing MAbs bound with high affinity to domains I or III, indicating that both affinity and the precise epitope recognized by an antibody are important determinants of WNV neutralization.
authors.affiliation: Department of Microbiology, University of Pennsylvania, 225 Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, PA 19104, USA.
authors.forename: Melissa D
authors.initials: MD
authors.lastname: Sánchez
authors.forename: Theodore C
authors.initials: TC
authors.lastname: Pierson
authors.forename: Douglas
authors.initials: D
authors.lastname: McAllister
authors.forename: Sheri L
authors.initials: SL
authors.lastname: Hanna
authors.forename: Bridget A
authors.initials: BA
authors.lastname: Puffer
authors.forename: Laura E
authors.initials: LE
authors.lastname: Valentine
authors.forename: Mariam M
authors.initials: MM
authors.lastname: Murtadha
authors.forename: James A
authors.initials: JA
authors.lastname: Hoxie
authors.forename: Robert W
authors.initials: RW
authors.lastname: Doms
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D003429:Cross Reactions; D004664:Encephalitis Virus, Japanese / Q000276:immunology; D004665:Encephalitis Virus, St. Louis / Q000276:immunology; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D006023:Glycoproteins / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D014902:West Nile virus / Q000276:immunology*; D015005:Yellow fever virus / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins