Antibody recognition and neutralization determinants on domains I and II of West Nile Virus envelope protein.
Previous studies have demonstrated that monoclonal antibodies (MAbs) against an epitope on the lateral surface of domain III (DIII) of the West Nile virus (WNV) envelope (E) strongly protect against infection in animals. Herein, we observed significantly less efficient neutralization by 89 MAbs that recognized domain I (DI) or II (DII) of WNV E protein. Moreover, in cells expressing Fc gamma receptors, many of the DI- and DII-specific MAbs enhanced infection over a broad range of concentrations. Using yeast surface display of E protein variants, we identified 25 E protein residues to be critical for recognition by DI- or DII-specific neutralizing MAbs. These residues cluster into six novel and one previously characterized epitope located on the lateral ridge of DI, the linker region between DI and DIII, the hinge interface between DI and DII, and the lateral ridge, central interface, dimer interface, and fusion loop of DII. Approximately 45% of DI-DII-specific MAbs showed reduced binding with mutations in the highly conserved fusion loop in DII: 85% of these (34 of 40) cross-reacted with the distantly related dengue virus (DENV). In contrast, MAbs that bound the other neutralizing epitopes in DI and DII showed no apparent cross-reactivity with DENV E protein. Surprisingly, several of the neutralizing epitopes were located in solvent-inaccessible positions in the context of the available pseudoatomic model of WNV. Nonetheless, DI and DII MAbs protect against WNV infection in mice, albeit with lower efficiency than DIII-specific neutralizing MAbs.
Oliphant
Theodore
T
Department of Medicine, Washington University School of Medicine, 660 South Euclid Ave., Box 8051, Saint Louis, MO 63110, USA.
Nybakken
Grant E
GE
Engle
Michael
M
Xu
Qing
Q
Nelson
Christopher A
CA
Sukupolvi-Petty
Soila
S
Marri
Anantha
A
Lachmi
Bat-El
BE
Olshevsky
Udy
U
Fremont
Daved H
DH
Pierson
Theodore C
TC
Diamond
Michael S
MS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics*; D051379:Mice; D009154:Mutation / Q000235:genetics; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary / Q000235:genetics; D017452:Receptors, IgG / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*; D014902:West Nile virus / Q000235:genetics* / Q000276:immunology; D015003:Yeasts
D000911:Antibodies, Monoclonal; D000939:Epitopes; D017452:Receptors, IgG; D014759:Viral Envelope Proteins