West Nile virus (WNV), a member of the Flavivirus genus, is a leading cause of viral encephalitis in the United States1. The development of neutralizing antibodies against the flavivirus envelope (E) protein is critical for immunity and vaccine protection2. Previously identified candidate therapeutic mouse and human neutralizing monoclonal antibodies (mAbs) target epitopes within the E domain III lateral ridge and the domain I-II hinge region, respectively3. To explore the neutralizing antibody repertoire elicited by WNV infection for potential therapeutic application, we isolated ten mAbs from WNV-infected individuals. mAb WNV-86 neutralized WNV with a 50% inhibitory concentration of 2 ng ml-1, one of the most potently neutralizing flavivirus-specific antibodies ever isolated. WNV-86 targets an epitope in E domain II, and preferentially recognizes mature virions lacking an uncleaved form of the chaperone protein prM, unlike most flavivirus-specific antibodies4. In vitro selection experiments revealed a neutralization escape mechanism involving a glycan addition to E domain II. Finally, a single dose of WNV-86 administered two days post-infection protected mice from lethal WNV challenge. This study identifies a highly potent human neutralizing mAb with therapeutic potential that targets an epitope preferentially displayed on mature virions.
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Additional data sets generated and/or analysed during the current study are available from the corresponding authors on reasonable request.
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Reimann, C. A. et al. Epidemiology of neuroinvasive arboviral disease in the United States, 1999–2007. Am. J. Trop. Med. Hyg. 79, 974–979 (2008).
Rey, F. A., Stiasny, K., Vaney, M. C., Dellarole, M. & Heinz, F. X. The bright and the dark side of human antibody responses to flaviviruses: lessons for vaccine design. EMBO Rep. 19, 206–224 (2018).
VanBlargan, L. A., Goo, L. & Pierson, T. C. Deconstructing the antiviral neutralizing-antibody response: implications for vaccine development and immunity. Microbiol. Mol. Biol. Rev. 80, 989–1010 (2016).
Nelson, S. et al. Maturation of West Nile virus modulates sensitivity to antibody-mediated neutralization. PLoS Pathog. 4, e1000060 (2008).
Pierson, T. C. & Diamond, M. S. Degrees of maturity: the complex structure and biology of flaviviruses. Curr. Opin. Virol. 2, 168–175 (2012).
Cherrier, M. V. et al. Structural basis for the preferential recognition of immature flaviviruses by a fusion-loop antibody. EMBO J. 28, 3269–3276 (2009).
Guirakhoo, F., Bolin, R. A. & Roehrig, J. T. The Murray Valley encephalitis virus prM protein confers acid resistance to virus particles and alters the expression of epitopes within the R2 domain of E glycoprotein. Virology 191, 921–931 (1992).
Heinz, F. X. et al. Structural changes and functional control of the tick-borne encephalitis virus glycoprotein E by the heterodimeric association with protein prM. Virology 198, 109–117 (1994).
Oliphant, T. et al. Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus. Nat. Med. 11, 522–530 (2005).
Throsby, M. et al. Isolation and characterization of human monoclonal antibodies from individuals infected with West Nile virus. J. Virol. 80, 6982–6992 (2006).
Gould, L. H. et al. Protective and therapeutic capacity of human single-chain Fv-Fc fusion proteins against West Nile virus. J. Virol. 79, 14606–14613 (2005).
Pierson, T. C. et al. The stoichiometry of antibody-mediated neutralization and enhancement of West Nile virus infection. Cell Host Microbe 1, 135–145 (2007).
Nybakken, G. E. et al. Structural basis of West Nile virus neutralization by a therapeutic antibody. Nature 437, 764–769 (2005).
Chung, W. M. et al. The 2012 West Nile encephalitis epidemic in Dallas, Texas. JAMA 310, 297–307 (2013).
Pierson, T. C. et al. A rapid and quantitative assay for measuring antibody-mediated neutralization of West Nile virus infection. Virology 346, 53–65 (2006).
Li, L. et al. The flavivirus precursor membrane-envelope protein complex: structure and maturation. Science 319, 1830–1834 (2008).
Dejnirattisai, W. et al. A new class of highly potent, broadly neutralizing antibodies isolated from viremic patients infected with dengue virus. Nat. Immunol. 16, 170–177 (2015).
Kaufmann, B. et al. Neutralization of West Nile virus by cross-linking of its surface proteins with Fab fragments of the human monoclonal antibody CR4354. Proc. Natl Acad. Sci. USA 107, 18950–18955 (2010).
Teoh, E. P. et al. The structural basis for serotype-specific neutralization of dengue virus by a human antibody. Sci. Transl Med. 4, 139ra183 (2012).
de Alwis, R. et al. Identification of human neutralizing antibodies that bind to complex epitopes on dengue virions. Proc. Natl Acad. Sci. USA 109, 7439–7444 (2012).
Hatcher, E. L. et al. Virus Variation Resource—improved response to emergent viral outbreaks. Nucleic Acids Res. 45, D482–D490 (2017).
Yu, X., McGraw, P. A., House, F. S. & Crowe, J. E. Jr. An optimized electrofusion-based protocol for generating virus-specific human monoclonal antibodies. J. Immunol. Methods 336, 142–151 (2008).
Davis, C. W. et al. West Nile virus discriminates between DC-SIGN and DC-SIGNR for cellular attachment and infection. J. Virol. 80, 1290–1301 (2006).
Ansarah-Sobrinho, C., Nelson, S., Jost, C. A., Whitehead, S. S. & Pierson, T. C. Temperature-dependent production of pseudoinfectious dengue reporter virus particles by complementation. Virology 381, 67–74 (2008).
Dowd, K. A. et al. Broadly neutralizing activity of Zika virus-immune sera identifies a single viral serotype. Cell Rep. 16, 1485–1491 (2016).
Lin, T. Y. et al. A novel approach for the rapid mutagenesis and directed evolution of the structural genes of West Nile virus. J. Virol. 86, 3501–3512 (2012).
Oliphant, T. et al. Induction of epitope-specific neutralizing antibodies against West Nile virus. J. Virol. 81, 11828–11839 (2007).
This study was funded by NIH grants R01 AI073755 (to M.S.D. and J.E.C.) and HHSN272201400018C (to M.S.D.), and by the intramural research program of the Division of Intramural Research, National Institutes of Allergy and Infectious Diseases (T.C.P.). Flow cytometry experiments were performed in the VMC Flow Cytometry Shared Resource, which is supported by the Vanderbilt Ingram Cancer Center (P30 CA68485) and the Vanderbilt Digestive Disease Research Center (DK058404). The project was supported by CTSA award no. UL1TR000445 from the National Center for Advancing Translational Sciences. Its contents are solely the responsibility of the authors and do not necessarily represent official views of the National Center for Advancing Translational Sciences or the National Institutes of Health.