title: Identification and characterization of antibody-binding epitopes on the norovirus GII.3 capsid.
abstract: Genotype II.3 (GII.3) noroviruses are a major cause of sporadic gastroenteritis, particularly in children. The greater incidence of GII.3 noroviruses in the pediatric population compared to the adult demographic suggests development of herd immunity to this genotype, possibly as a consequence of limited evolution of immune epitopes. This study aimed to identify and characterize immune epitopes on the GII.3 capsid protein and to determine the level of immune cross-reactivity within the genotype. A panel of seven GII.3 virus-like particles (VLPs), representing norovirus strains isolated during 1975 to 2008, was tested by enzyme-linked immunosorbent assay (ELISA) for reactivity with human sera and a rabbit anti-GII.3 strain-specific polyclonal serum generated against the 2008 GII.3 VLP. Immunoprecipitation of protease-digested GII.3 VLPs and sequencing of bound peptides via mass spectrometry were used to locate epitopes on the capsid. Two epitopes were investigated further using Mimotopes technology. Serum binding studies demonstrated complete intragenotype GII.3 cross-reactivity using both human and rabbit serum. Six immunoreactive regions containing epitopes were located on the GII.3 capsid protein, two within each capsid domain. Epitopes in the S and P1 domains were highly conserved within GII.3 noroviruses. P2 domain epitopes were variable and contained evolutionarily important residues and histo-blood group antigen (HBGA) binding residues. In conclusion, anti-GII.3 antibody-binding epitopes are highly cross-reactive and mostly conserved within GII.3 strains. This may account for the limited GII.3 prevalence in adults and suggests that a GII.3 strain may be a valuable inclusion in a multivalent pediatric targeted VLP vaccine. Exploration of norovirus immune epitopes is vital for effective vaccine design. IMPORTANCE This study represents an important contribution to the understanding of norovirus immunology in a pediatric genotype. The high cross-reactivity and conservation of GII.3 epitopes suggest development of herd immunity against GII.3 and indicate that a GII.3 strain would be a valuable inclusion in a pediatric targeted multivalent vaccine. Immunological understanding of pediatric norovirus strains is important since norovirus vaccines will likely target high-risk groups such as the pediatric population.
authors.affiliation: Enteric Virus Group, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, VIC, Australia.
authors.forename: Jackie E
authors.initials: JE
authors.lastname: Mahar
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Donker
authors.forename: Karin
authors.initials: K
authors.lastname: Bok
authors.forename: Gert H
authors.initials: GH
authors.lastname: Talbo
authors.forename: Kim Y
authors.initials: KY
authors.lastname: Green
authors.forename: Carl D
authors.initials: CD
authors.lastname: Kirkwood
mesh_terms: D000595:Amino Acid Sequence; D000704:Analysis of Variance; D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism*; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002853:Chromatography, Liquid; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005759:Gastroenteritis / Q000276:immunology* / Q000821:virology*; D006801:Humans; D032521:Immunity, Herd / Q000276:immunology*; D047468:Immunoprecipitation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D029322:Norovirus / Q000235:genetics* / Q000276:immunology; D011817:Rabbits; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D053719:Tandem Mass Spectrometry
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes