title: Multiple antigenic sites are involved in blocking the interaction of GII.4 norovirus capsid with ABH histo-blood group antigens.
abstract: Noroviruses are major etiological agents of acute viral gastroenteritis. In 2002, a GII.4 variant (Farmington Hills cluster) spread so rapidly in the human population that it predominated worldwide and displaced previous GII.4 strains. We developed and characterized a panel of six monoclonal antibodies (MAbs) directed against the capsid protein of a Farmington Hills-like GII.4 norovirus strain that was associated with a large hospital outbreak in Maryland in 2004. The six MAbs reacted with high titers against homologous virus-like particles (VLPs) by enzyme-linked immunoassay but did not react with denatured capsid protein in immunoblots. The expression and self-assembly of newly developed genogroup I/II chimeric VLPs showed that five MAbs bound to the GII.4 protruding (P) domain of the capsid protein, while one recognized the GII.4 shell (S) domain. Cross-competition assays and mutational analyses showed evidence for at least three distinct antigenic sites in the P domain and one in the S domain. MAbs that mapped to the P domain but not the S domain were able to block the interaction of VLPs with ABH histo-blood group antigens (HBGA), suggesting that multiple antigenic sites of the P domain are involved in HBGA blocking. Further analysis showed that two MAbs mapped to regions of the capsid that had been associated with the emergence of new GII.4 variants. Taken together, our data map antibody and HBGA carbohydrate binding to proximal regions of the norovirus capsid, showing that evolutionary pressures on the norovirus capsid protein may affect both antigenic and carbohydrate recognition phenotypes.
authors.affiliation: Caliciviruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, DHHS, Bethesda, Maryland, USA.
authors.forename: Gabriel I
authors.initials: GI
authors.lastname: Parra
authors.forename: Eugenio J
authors.initials: EJ
authors.lastname: Abente
authors.forename: Carlos
authors.initials: C
authors.lastname: Sandoval-Jaime
authors.forename: Stanislav V
authors.initials: SV
authors.lastname: Sosnovtsev
authors.forename: Karin
authors.initials: K
authors.lastname: Bok
authors.forename: Kim Y
authors.initials: KY
authors.lastname: Green
mesh_terms: D000017:ABO Blood-Group System / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D017250:Caliciviridae Infections / Q000453:epidemiology / Q000821:virology*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D004196:Disease Outbreaks; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005838:Genotype; D006801:Humans; D008396:Maryland / Q000453:epidemiology; D051379:Mice; D008807:Mice, Inbred BALB C; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D011485:Protein Binding; D025941:Protein Interaction Mapping
chemical_list: D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins