title: Antigenic structure of human hepatitis A virus defined by analysis of escape mutants selected against murine monoclonal antibodies.
abstract: We examined the antigenic structure of human hepatitis A virus (HAV) by characterizing a series of 21 murine monoclonal-antibody-resistant neutralization escape mutants derived from the HM175 virus strain. The escape phenotype of each mutant was associated with reduced antibody binding in radioimmunofocus assays. Neutralization escape mutations were identified at the Asp-70 and Gln-74 residues of the capsid protein VP3, as well as at Ser-102, Val-171, Ala-176, and Lys-221 of VP1. With the exception of the Lys-221 mutants, substantial cross-resistance was evident among escape mutants tested against a panel of 22 neutralizing monoclonal antibodies, suggesting that the involved residues contribute to epitopes composing a single antigenic site. As mutations at one or more of these residues conferred resistance to 20 of 22 murine antibodies, this site appears to be immunodominant in the mouse. However, multiple mutants selected independently against any one monoclonal antibody had mutations at only one or, at the most, two amino acid residues within the capsid proteins, confirming that there are multiple epitopes within this antigenic site and suggesting that single-amino-acid residues contributing to these epitopes may play key roles in the binding of individual antibodies. A second, potentially independent antigenic site was identified by three escape mutants with different substitutions at Lys-221 of VP1. These mutants were resistant only to antibody H7C27, while H7C27 effectively neutralized all other escape mutants. These data support the existence of an immunodominant neutralization site in the antigenic structure of hepatitis A virus which involves residues of VP3 and VP1 and a second, potentially independent site involving residue 221 of VP1.
authors.affiliation: Department of Medicine, University of North Carolina, Chapel Hill 27599-7030.
authors.forename: L H
authors.initials: LH
authors.lastname: Ping
authors.forename: S M
authors.initials: SM
authors.lastname: Lemon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D003429:Cross Reactions; D006507:Hepatovirus / Q000276:immunology*; D051379:Mice; D009154:Mutation; D009500:Neutralization Tests
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral