Authors: NguyenXuan Huy SaeHae Kim MoonSik Yang TaeGeum Kim
Publish Date: 2012/06/27
Volume: 31, Issue: 10, Pages: 1933-1942
Abstract
To increase immune responses of plantbased vaccines in intestine mucosal immune systems a synthetic neutralizing epitope sCOE gene of porcine epidemic diarrhea virus PEDV was fused with M celltargeting ligand Co1 and introduced into a plant expression vector under the control of rice amylase 3D promoter The sCOE–Co1 fusion gene was introduced into rice calli via the particle bombardmentmediated transformation method The stable integration and transcriptional expression of the sCOE–Co1 fusion gene was confirmed by genomic DNA PCR amplification and Northern blot analysis respectively The expression of the COE–Co1 fusion protein was confirmed by immunoblot analysis The highest expression level of the COE–Co1 fusion protein reached 0083 of the total soluble protein according to quantitative densitometry of Western blot analysis Mice immunized with transgenic rice calli protein extracts induced significant serum IgG and fecal IgA antibody levels against purified bacterial COE The systemic and mucosal immune responses were confirmed by measuring COEspecific IgG and IgA antibodysecreting cells in the lymphocytes extracted from the spleen and COEspecific IgA antibodysecreting cells in the Peyer’s patches from immunized mice These results indicated that oral immunization of plantproduced COE–Co1 fusion protein could elicit efficient systemic and mucosal immune responses against the COE antigen
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