Isolation and characterization of human monoclonal antibodies from individuals infected with West Nile Virus.
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.
Throsby
Mark
M
Crucell Holland B.V., P.O. Box 2048, 2301 CA, Leiden, The Netherlands. m.throsby@crucell.com
Geuijen
Cecile
C
Goudsmit
Jaap
J
Bakker
Arjen Q
AQ
Korimbocus
Jehanara
J
Kramer
R Arjen
RA
Clijsters-van der Horst
Marieke
M
de Jong
Maureen
M
Jongeneelen
Mandy
M
Thijsse
Sandra
S
Smit
Renate
R
Visser
Therese J
TJ
Bijl
Nora
N
Marissen
Wilfred E
WE
Loeb
Mark
M
Kelvin
David J
DJ
Preiser
Wolfgang
W
ter Meulen
Jan
J
de Kruif
John
J
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*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins