N-linked glycosylation alteration in E0 and E2 glycoprotein of classical swine fever virus and novel classical swine fever virus vaccine

Patent metadata

Field	Value
Publication number	US-8961996-B2
Application number	US-91332910-A
Country	US
Kind code	B2
Filing date	Oct 27, 2010
Priority date	Oct 24, 2008
Publication date	Feb 24, 2015
Grant date	Feb 24, 2015

Abstract

E2 is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). 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. 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 post-infection suggesting that glycosylation of E2 could be modified for development of CSF live-attenuated vaccines. Additionally, a new developed virus, contained deletions of putative glycosylation sites N1 in E2 and N1 in E0 (6b), called N1E0/2v, induce a solid protection against the challenge at 3 and 28 days post-inoculation.

First claim

We claim:

1. A genetically modified recombinant classical swine fever virus (CSFV) mutant of a highly pathogenic native Brescia strain, said mutant, N1E0/2v, comprising a cDNA encoding two mutations, wherein one of the mutations is in CSFV E0 glycoprotein, which is presented in the N1 site of the E0 glycoprotein, and the glycosylated amino acid asparagine at position 269 of the E0 glycoprotein is altered to the non-glycosylated amino acid, alanine; and another of the mutations is i…

Assignees

• Borca Manuel
• Risatti Guillermo
• Us Agriculture
• Univ Connecticut

Inventors

• Borca Manuel
• Risatti Guillermo

Classifications

• C12Q1/701

  Chemistry & Metallurgy · mapped topic

• G01N2333/183

  Physics · mapped topic

• C12N2770/24334

  Chemistry & Metallurgy · mapped topic

• C12N2770/24322

  Chemistry & Metallurgy · mapped topic

• A61K39/12

  Human Necessities · mapped topic