title: Isolation and characterization of human monoclonal antibodies from individuals infected with West Nile Virus.
abstract: Monoclonal antibodies (MAbs) neutralizing West Nile Virus (WNV) have been shown to protect against infection in animal models and have been identified as a correlate of protection in WNV vaccine studies. In the present study, antibody repertoires from three convalescent WNV-infected patients were cloned into an scFv phage library, and 138 human MAbs binding to WNV were identified. One hundred twenty-one MAbs specifically bound to the viral envelope (E) protein and four MAbs to the premembrane (prM) protein. Enzyme-linked immunosorbent assay-based competitive-binding assays with representative E protein-specific MAbs demonstrated that 24/51 (47%) bound to domain II while only 4/51 (8%) targeted domain III. In vitro neutralizing activity was demonstrated for 12 MAbs, and two of these, CR4374 and CR4353, protected mice from lethal WNV challenge at 50% protective doses of 12.9 and 357 mug/kg of body weight, respectively. Our data analyzing three infected individuals suggest that the human anti-WNV repertoire after natural infection is dominated by nonneutralizing or weakly neutralizing MAbs binding to domain II of the E protein, while domain III-binding MAbs able to potently neutralize WNV in vitro and in vivo are rare.
authors.affiliation: Crucell Holland B.V., P.O. Box 2048, 2301 CA, Leiden, The Netherlands. m.throsby@crucell.com
authors.forename: Mark
authors.initials: M
authors.lastname: Throsby
authors.forename: Cecile
authors.initials: C
authors.lastname: Geuijen
authors.forename: Jaap
authors.initials: J
authors.lastname: Goudsmit
authors.forename: Arjen Q
authors.initials: AQ
authors.lastname: Bakker
authors.forename: Jehanara
authors.initials: J
authors.lastname: Korimbocus
authors.forename: R Arjen
authors.initials: RA
authors.lastname: Kramer
authors.forename: Marieke
authors.initials: M
authors.lastname: Clijsters-van der Horst
authors.forename: Maureen
authors.initials: M
authors.lastname: de Jong
authors.forename: Mandy
authors.initials: M
authors.lastname: Jongeneelen
authors.forename: Sandra
authors.initials: S
authors.lastname: Thijsse
authors.forename: Renate
authors.initials: R
authors.lastname: Smit
authors.forename: Therese J
authors.initials: TJ
authors.lastname: Visser
authors.forename: Nora
authors.initials: N
authors.lastname: Bijl
authors.forename: Wilfred E
authors.initials: WE
authors.lastname: Marissen
authors.forename: Mark
authors.initials: M
authors.lastname: Loeb
authors.forename: David J
authors.initials: DJ
authors.lastname: Kelvin
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Preiser
authors.forename: Jan
authors.initials: J
authors.lastname: ter Meulen
authors.forename: John
authors.initials: J
authors.lastname: de Kruif
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology; D001666:Binding Sites, Antibody / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D006801:Humans; D051379:Mice; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000276:immunology*; D014901:West Nile Fever / Q000276:immunology*; D014902:West Nile virus / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins