Immunoprotective properties of transgenic plants expressing E2 glycoprotein from CSFV and cysteine protease from Fasciola hepatica
Introduction
Research on plant-derived, orally administrated vaccines has been possible further to the development of effective methods of plant transformation and the identification of highly immunogenic epitopes from pathogens [1], [2], [3]. The aim of this work was to obtain transgenic plants expressing the E2 glycoprotein from Classical Swine Fever Virus (CSFV) or the antigenic cysteine protease from Fasciola hepatica, and to test the ability of the recombinant antigens to induce an antibody response in laboratory animals when administered orally.
Classical Swine Fever Virus, also termed Hog Cholera Virus, belongs to the Pestiviridae genus of the Flaviviridae family [4], and is the causative agent of a contagious, often fatal, pig disease. Among the proteins encoded by the CSFV genome, the glycosylated family of E proteins, with protein E2 (former name gp51–54 according to the apparent molecular weight), is considered as highly antigenic. We sequenced the E2 CSFV coding region from two viral strains: Brescia (1526 bp) and Alfort (1173 bp), and used the former sequence for the construction of transformation vectors described in these studies.
F. hepatica, the liver fluke, causes widespread disease in farm animals as well as in humans. Fasciolosis is a chronic helminthes disease with a worldwide distribution and is a major cause of morbidity and mortality in domestic ruminants (cattle, sheep, goats), causing severe economic losses. Humans may also acquire the infection, as reported from several developing countries. Among the proteins released by flukes, there are potent proteases essential for parasite metabolism. Preliminary reports indicated that cysteine proteases might be a source of protective antigens [5], [6]. A 981 nt cDNA fragment encoding the cysteine protease from F. hepatica was used for the construction of transformation vectors for production of the protein in plants. The immunizing potential of this antigen administered orally was tested in laboratory animals.
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Results and discussion
Two series of plant transformation vectors were constructed: the first one with the coding sequence for the E2 glycoprotein of CSFV and the second one with the leader (coding for 96 amino acids) or catalytic domain (coding for 223 amino acid fragment) of the cysteine protease of F. hepatica (Fig. 1). Some vectors were equipped with VTS (vacuolar target) or KDEL (protein retention in endoplasmic reticulum) sequences. Antigen-coding sequences were coupled with hepatitis B virus (HBV) core protein
Acknowledgment
This work was financed by PBZ-MIN-007/P04/2003 Grant from Polish Ministry of Scientific Research and Information Technology.
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