Expression and Immunological Studies of Classical Swine Fever Virus Glycoprotein E2 in the Bi-Cistronic Baculovirus/Larvae Expression System

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2010, Vol.74 (7), p.1343-1349
• Main Authors: WU, Chi-Ming, HSUAN, Shih-Ling, CHEN, Zeng-Weng, JINN, Tzyy-Rong, HUANG, Chienjin, LIAO, Jiunn-Wang, CHEN, Ter-Hsin, LIAO, Chih-Ming, LEE, Wei-Cheng, WU, Tzong-Yuan, CHIEN, Maw-Sheng
• Format: Article
• Language: English
• Subjects: Animals; Baculoviridae - genetics; Baculovirus; bi-cistronic; Biological and medical sciences; Classical swine fever virus; classical swine fever virus (CSFV) E2 glycoprotein; Fundamental and applied biological sciences. Psychology; Gene Expression; green fluorescent protein (GFP); Immunization; Larva - genetics; Lepidoptera - cytology; Lepidoptera - genetics; Male; Mice; Peptide Hydrolases - metabolism; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Recombinant Proteins - metabolism; Trichoplusia ni; Trichoplusia ni larvae; Viral Envelope Proteins - biosynthesis; Viral Envelope Proteins - genetics; Viral Envelope Proteins - immunology; Viral Envelope Proteins - metabolism
• ISSN: 0916-8451; 1347-6947; 1347-6947
• DOI: 10.1271/bbb.90894

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.