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Isolation and Characterization of Human Monoclonal Antibodies from Individuals Infected with West Nile Virus

Mark Throsby,^1* Cecile Geuijen,¹ Jaap Goudsmit,¹ Arjen Q. Bakker,¹ Jehanara Korimbocus,² R. Arjen Kramer,¹ Marieke Clijsters-van der Horst,¹ Maureen de Jong,¹ Mandy Jongeneelen,¹ Sandra Thijsse,¹ Renate Smit,¹ Therese J. Visser,¹ Nora Bijl,^1, Wilfred E. Marissen,¹ Mark Loeb,³ David J. Kelvin,⁴ Wolfgang Preiser,^5, Jan ter Meulen,¹ and John de Kruif¹

Crucell Holland B.V., Leiden, The Netherlands,¹ National Reference Center for Arbovirus and Viral Hemorrhagic Fevers, Pasteur Institute, Lyon, France,² Department of Clinical Epidemiology and Biostatistics, McMaster University, Hamilton, Canada,³ Division of Experimental Therapeutics, University Health Network, Toronto, Canada,⁴ Institute of Medical Virology, W. Goethe-Universität, Frankfurt, Germany⁵

Received 16 March 2006/ Accepted 21 April 2006

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 µg/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.

Supplemental material for this article may be found at http://jvi.asm.org.

Present address: Department of Medical Biochemistry, Academic Medical Centre, Amsterdam, The Netherlands.

Present address: Department of Pathology, University of Stellenbosch, Tygerberg, South Africa.

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