title: Fine mapping of a salmonid E2 alphavirus neutralizing epitope.
abstract: In this study, we aimed to characterize the epitope recognized by the neutralizing 17H23 mAb directed against the E2 glycoprotein of most of salmonid alphavirus (SAV) subtypes and widely used in several laboratories to routinely diagnose SAV. We hypothesized that the 17H23 epitope was located in the major domain B, previously identified in the E2 of mammalian alphaviruses as the domain recognized by most of the E2 neutralizing mAbs. Indeed, the SAV E2 domain B counterpart is contained in the protein domain previously characterized as being recognized by mAb 17H23. Thus, to precisely characterize the 17H23 epitope, we developed an alanine scanning mutagenesis approach coupled with the generation of the respective recombinant SAV (rSAV) by using the available infectious cDNA. Ten mutant rSAVs termed A-J from E2 aa 223-236 were produced and characterized in vitro using indirect immunofluorescence assays on virus-infected cells with mAbs 17H23, 51B8 (another non-neutralizing anti-E2 mAb) and 19F3 directed against the non-structural protein nsp1. Two of the mutant rSAVs (G and H) escaped neutralization by mAb 17H23. In addition, we showed that when juvenile trout were infected by bath immersion with the rSAV mutants, some of them were either totally (D, E and G) or partially (H) attenuated. Together, the data from the in vitro and in vivo experiments indicated that the putative 17H23 amino acid sequence epitope comprised the short amino acid sequence (227)FTSDS(231).
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Emilie
authors.initials: E
authors.lastname: Mérour
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Annie
authors.initials: A
authors.lastname: Lamoureux
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Biacchesi
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Brémont
mesh_terms: D000524:Alphavirus / Q000235:genetics / Q000276:immunology*; D018354:Alphavirus Infections / Q000235:genetics / Q000276:immunology* / Q000821:virology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology*; D002460:Cell Line; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D005393:Fish Diseases / Q000235:genetics / Q000276:immunology* / Q000821:virology; D015870:Gene Expression; D054884:Host-Pathogen Interactions; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D017686:Oncorhynchus mykiss / Q000821:virology; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins