title: Alternative codon usage of PRRS virus ORF5 gene increases eucaryotic expression of GP(5) glycoprotein and improves immune response in challenged pigs.
abstract: Pigs exposed to GP(5) protein of PRRSV by means of DNA immunization develop specific neutralizing and protecting antibodies. Herein, we report on the consequences of codon bias, and on the favorable outcome of the systematic replacement of native codons of PRRSV ORF5 gene with codons chosen to reflect more closely the codon preference of highly expressed mammalian genes. Therefore, a synthetic PRRSV ORF5 gene (synORF5) was constructed in which 134 nucleotide substitutions were made in comparison to wild-type gene (wtORF5), such that 59% (119) of wild-type codons were replaced with known preferable codons in mammalian cells. In vitro expression in mammalian cells of synORF5 was considerably increased comparatively to wtORF5, following infection with tetracycline inducible replication-defective human adenoviral vectors (hAdVs). After challenge inoculation, SPF pigs vaccinated twice with recombinant hAdV/synORF5 developed earlier and higher antibody titers, including virus neutralizing antibodies to GP(5) than pigs vaccinated with hAdV/wtORF5. Data obtained from animal inoculation studies suggest direct correlation between expression levels of immunogenic structural viral proteins and immune response.
authors.affiliation: INRS-Institut Armand-Frappier, 531 boul. des Prairies, Laval, P.Q., Canada H7V 1B7.
authors.forename: Ali
authors.initials: A
authors.lastname: Kheyar
authors.forename: Ahmed
authors.initials: A
authors.lastname: Jabrane
authors.forename: Chengru
authors.initials: C
authors.lastname: Zhu
authors.forename: Patrick
authors.initials: P
authors.lastname: Cléroux
authors.forename: Bernard
authors.initials: B
authors.lastname: Massie
authors.forename: Serge
authors.initials: S
authors.lastname: Dea
authors.forename: Carl A
authors.initials: CA
authors.lastname: Gagnon
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D003062:Codon; D006801:Humans; D009500:Neutralization Tests; D016366:Open Reading Frames; D019318:Porcine Reproductive and Respiratory Syndrome / Q000517:prevention & control*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000276:immunology*; D013552:Swine; D019444:Vaccines, DNA / Q000008:administration & dosage* / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D003062:Codon; D019444:Vaccines, DNA; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C114789:glycoprotein 5, PRRSV