Evaluation of Adjuvant Activity of Heat Shock Protein Fused to FMD Virus Polyprotein in Development of Mucosal Vaccine
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Date
2009-07-17
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Karnataka Veterinary, Animal and Fisheries Sciences University , Bidar
Abstract
The present work was undertaken to develop a mucosal vaccine as a therapeutic vaccine against FMD on the hypothesis that, the delivered candidate vaccine, if made anchored on mucosal surface may stimulate potent-IgA response, which can neutralize the virus during spread of the disease. In our present study, we made use of Enolase gene isolated from oral commensal of cattle, Enterococcus faecalis and FMDV polyvalent protein gene encoding major immunogenic epitopes of three serotypes (A22, Asia-1 and ‘O’).
Enolase, a heat shock protein is identified to bind to plasminogen and to mucosal surface mainly by lysine residues in the C-terminal of the protein. α Enolase may be an excellent adapter molecule in the application of antigen delivery at the mucosal surface. The gene Enolase was used to produce a fusion protein with FMDV immunogen at the C-terminal end. For this, FMDV polyvalent protein gene was inserted at Not1 and Xho1 sites in the pET-32a+ vector. Enolase was inserted at Bam H1 and Xho1 sites of the pET-32a+ vector so as to get FMDV polyvalent protein gene at the 3’ end of Enolase. The construct was induced to get 3’fusion protein which was confirmed by immunoblot assay. The 3’fusion protein after purification was used for immune response study in guinea pigs. Oral wash and serum samples of these guinea pigs subjected to double sandwich ELISA showed that there was a specific IgA production.
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P.G. Thesis
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