Antigenicity of the N8 influenza A virus neuraminidase: existence of an epitope at the subunit interface of the neuraminidase

Abstract

To locate antigenic epitopes on the N8 neuraminidase (NA), we generated a panel of 97 monoclonal antibodies (MAbs), 66 of which inhibited NA activity (NI antibodies). Three groups of NI MAbs were identified from their different reactivities with escape mutants. Group 1 antibodies recognized the peptide loop containing residues 344 to 346, which appears to be an immunodominant region on the rim of the enzyme center of the N8 NA. Group 2 antibodies recognized a novel epitope containing residues 150, 199, 367, 399, and 400 (N2 numbering). From the location of these residues on the three-dimensional structure of the N8 NA, the epitope appears to be located at the interface of two adjacent monomers in the tetrameric NA, one contributing residues 150 and 199 and the other contributing residues 367 and 399 to 400. The available evidence indicates that the MAbs of this group react with the NA only after it is fully assembled. The third group of antibodies recognized the peptide loops containing residues 367 and 399 to 400. All of the amino acid substitutions in N8 escape mutants which affect the NI activity of antibodies were located in the peptide loops known to form epitopes in the N2 and N9 subtypes, indicating that antigenic regions in the NA head inducing NI antibodies appear to be similar among different subtypes of influenza A viruses. The MAbs used in this study will be valuable in studying the role of each N8 NA epitope in host immune defense systems and in the kinetics analysis of the biosynthesis of the enzyme.