title: N-linked glycosylation status of classical swine fever virus strain Brescia E2 glycoprotein influences virulence in swine.
abstract: E2 is one of the three envelope glycoproteins of classical swine fever virus (CSFV). Previous studies indicate that E2 is involved in several functions, including virus attachment and entry to target cells, production of antibodies, induction of protective immune response in swine, and virulence. Here, we have investigated the role of E2 glycosylation of the highly virulent CSFV strain Brescia in infection of the natural host. Seven putative glycosylation sites in E2 were modified by site-directed mutagenesis of a CSFV Brescia infectious clone (BICv). A panel of virus mutants was obtained and used to investigate whether the removal of putative glycosylation sites in the E2 glycoprotein would affect viral virulence/pathogenesis in swine. We observed that rescue of viable virus was completely impaired by removal of all putative glycosylation sites in E2 but restored when mutation N185A reverted to wild-type asparagine produced viable virus that was attenuated in swine. Single mutations of each of the E2 glycosylation sites showed that amino acid N116 (N1v virus) was responsible for BICv attenuation. N1v efficiently protected swine from challenge with virulent BICv at 3 and 28 days postinfection, suggesting that glycosylation of E2 could be modified for development of classical swine fever live attenuated vaccines.
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, PO Box 848, Greenport, NY 11944-0848, USA.
authors.forename: G R
authors.initials: GR
authors.lastname: Risatti
authors.forename: L G
authors.initials: LG
authors.lastname: Holinka
authors.forename: I
authors.initials: I
authors.lastname: Fernandez Sainz
authors.forename: C
authors.initials: C
authors.lastname: Carrillo
authors.forename: Z
authors.initials: Z
authors.lastname: Lu
authors.forename: M V
authors.initials: MV
authors.lastname: Borca
mesh_terms: D000818:Animals; D006691:Classical Swine Fever / Q000503:physiopathology* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D006031:Glycosylation; D009154:Mutation; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus