title: Expression and immunological studies of classical swine fever virus glycoprotein E2 in the bi-cistronic baculovirus/larvae expression system.
abstract: To develop an economical, easy technique for producing recombinant E2 glycoprotein (rE2) of classical swine fever virus (CSFV) as a candidate immunogen, a bi-cistronic baculovirus/larvae expression vector was constructed using p10 promoter, an internal ribosome entry site, and the gfp gene. Trichoplusia ni larvae were successfully infected with the occluded recombinant baculovirus via feed, and the characteristics of rE2 were confirmed by immunoblot and glycosylation stain. rE2 at a concentration of 0.6-0.8 mg/ml without degradation was obtained from hemolymphs of infected larvae that emitted high levels of green fluorescence. Immunization assays indicated that mice and piglets immunized with rE2-containing hemolymph elicited high titers of anti-CSFV E2 antibodies with virus-neutralizing activity. This is the first study to indicate that baculovirus/T. ni larvae-expressed rE2 can be served as a vaccine candidate. This system provides an economical alternative for the production of vaccine components in the veterinary industry.
authors.affiliation: Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University.
authors.forename: Chi-Ming
authors.initials: CM
authors.lastname: Wu
authors.forename: Shih-Ling
authors.initials: SL
authors.lastname: Hsuan
authors.forename: Zeng-Weng
authors.initials: ZW
authors.lastname: Chen
authors.forename: Tzyy-Rong
authors.initials: TR
authors.lastname: Jinn
authors.forename: Chienjin
authors.initials: C
authors.lastname: Huang
authors.forename: Jiunn-Wang
authors.initials: JW
authors.lastname: Liao
authors.forename: Ter-Hsin
authors.initials: TH
authors.lastname: Chen
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Liao
authors.forename: Wei-Cheng
authors.initials: WC
authors.lastname: Lee
authors.forename: Tzong-Yuan
authors.initials: TY
authors.lastname: Wu
authors.forename: Maw-Sheng
authors.initials: MS
authors.lastname: Chien
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D006692:Classical Swine Fever Virus; D015870:Gene Expression; D007114:Immunization; D007814:Larva / Q000235:genetics; D007915:Lepidoptera / Q000166:cytology / Q000235:genetics; D008297:Male; D051379:Mice; D010447:Peptide Hydrolases / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus; D010447:Peptide Hydrolases