Expression and immunological studies of classical swine fever virus glycoprotein E2 in the bi-cistronic baculovirus/larvae expression system.
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.
Wu
Chi-Ming
CM
Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University.
Hsuan
Shih-Ling
SL
Chen
Zeng-Weng
ZW
Jinn
Tzyy-Rong
TR
Huang
Chienjin
C
Liao
Jiunn-Wang
JW
Chen
Ter-Hsin
TH
Liao
Chih-Ming
CM
Lee
Wei-Cheng
WC
Wu
Tzong-Yuan
TY
Chien
Maw-Sheng
MS
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
D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus; D010447:Peptide Hydrolases