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Plant made veterinary vaccines against RHD
S44
Abstracts / Journal of Biotechnology 131S (2007) S43–S47
provides cross-protection against enterotoxic Escherichia coli. This vaccine is not available, however, in large part due to cost of production of the bacterial toxin protein which is a component of its formulation. The edible vaccines that at the moment are in clinical phases of research (Phase I) are the following ones: vaccine against the Enteritis caused by E. coli. expressed in potato plants; vaccine against infections caused by the virus Norwalk, expressed in potato plants and tobacco; vaccine against the virus of the Hepatitis B expressed in tobacco. Other fruits under experiment are banana and tomato. The latest research on topic are focused on multicomponent vaccines development that provide protection against several pathogens.
Mikschofsky, H., Schirrmeier, H., Broer, I., 2006. Towards plant made vaccines. In: Plant Genetics - Joined Conference of the German Genetics Society and the German Society for Plant Breeding, Kiel/Germany, 20–23 September 2006, p. 139.
doi:10.1016/j.jbiotec.2007.07.070
2 Friedrich-Loeffler-Institute
3. Plant made veterinary vaccines against RHD
The Rabbit hemorrhagic disease (RHD) is an acute and highly infectious viral disease affecting the European rabbit (Oryctolagus cuniculus). It causes high economic damages in rabbit meat production as well as limitations of wild populations. RHDvaccines are commercially produced by knowingly infected rabbits. VP60, the only structural capsid protein of RHDV, seems to be an appropriate subunit vaccine and offers possibilities to develop alternative vaccine production strategies. Plants demonstrate a promising production platform for vaccines, so far many different recombinant vaccines were expressed by higher plants -amongst others VP60 (Castanon et al., 2002; Gil et al., 2006). However, low expression levels combined with low immunogenicity of plant-derived VP60 prevented commercialisation of a plant-derived RHD-vaccine. Our aim was to enhance expression and immunogenicity of VP60 in different plant species in order to develop suitable plantderived RHD-vaccines. Different genetic modifications (codon adaptation, integration of regulatory and putative stabilizing sequences, and the addition of the well-known adjuvant ctb) led to a higher expression level as well to a tremendous enhancement of the immunogenicity of plant-derived VP60 (Mikschofsky et al., 2006). Surprisingly, only one of four synthetic ctbvp60 genes led to the expression of oligomers of pentameric CTB::VP60 in tobacco. This was only possible in tobacco and pea but not in potato, carrot or canola. Recombinant CTB::VP60 isolated from tobacco leaves demonstrated at least a 40fold higher immunogenicity compared to potato derived VP60-vaccines (Castanon et al., 2002; Castanon et al., 1999).
Heike Mikschofsky 1,∗ , Martin Hammer 2 , Patricia K¨onig 2 , G¨unther Keil 2 , Horst Schirrmeier 2 , Inge Broer 1 1 University
of Rostock, Rostock, Germany Insel, Riems, Germany
2 Friedrich-Loeffler-Institute
Vaccines against the rabbit hemorrhagic disease virus (RHDV), a highly infectious pathogen of the European rabbit (Oryctolagus cuniculus), are commercially produced by experimentally infected rabbits. VP60, the only structural capsid protein of RHDV, seems to be an appropriate subunit vaccine and offers possibilities to develop an alternative vaccine production strategy. Transgenic plants demonstrate a promising production platform for veterinary vaccines, up to now a number of viral and bacterial antigens were expressed by higher plants amongst others VP60 (Castanon et al., 2002; Gil et al., 2006). However, low expression levels and non-satisfying immunogenicity of plantderived VP60 prevented commercialisation of a plant-derived RHD-vaccine up to now. In order to develop suitable plant-derived RHD-vaccines we are studying the enhancement of the expression and the immunogenicity of VP60 in different plant species. Different genetic modifications (codon adaptation, integration of regulatory and putative stabilizing sequences, and the addition of the wellknown adjuvant ctb) led to a higher expression level as well as to a tremendous enhancement of the immunogenicity of plantderived VP60 (Mikschofsky et al., 2006). Interestingly, only one of four synthetic ctbvp60 genes led to the expression of oligomers of pentameric CTB::VP60. This was only possible in tobacco and pea but not in potato, carrot or canola. Tobacco and pea derived CTB::VP60 demonstrated at least a 40-fold higher immunogenicity compared to VP60-vaccines of potato tubers (Castanon et al., 1999, 2002). References Castanon, S., Martin-Alonso, J.M., Marin, M.S., Boga, J.A., Alonso, P., Parra, F., Ordas, R.J., 2002. Plant Sci. 162, 87–95. Castanon, S., Marin, M.S., Martin-Alonso, J.M., Boga, J.A., Casais, R., Humara, J.M., Ordas, R.J., Parra, F., 1999. J. Virol. 73, 4452–4455. Gil, F., Titarenko, E., Arcalis, E., Escribano, J.M., 2006. Plant Biotechnol. J. 4, 135–143.
doi:10.1016/j.jbiotec.2007.07.071 4. Production of high immunogenic VP60, a promising RHDvaccine, in plants Heike Mikschofsky 1,∗ , Martin Hammer 2 , Patricia K¨onig 2 , G¨unther Keil 2 , Horst Schirrmeier 2 , Inge Broer 1 1 University
of Rostock, Rostock, Germany Insel, Riems, Germany
References Castanon, S., Martin-Alonso, J.M., Marin, M.S., Boga, J.A., Alonso, P., Parra, F., Ordas, R.J., 2002. Plant Sci. 162, 87–95. Castanon, S., Marin, M.S., Martin-Alonso, J.M., Boga, J.A., Casais, R., Humara, J.M., Ordas, R.J., Parra, F., 1999. J. Virol. 73, 4452–4455. Gil, F., Titarenko, E., Arcalis, E., Escribano, J.M., 2006. Plant Biotechnol. J. 4, 135–143. Mikschofsky, H., Schirrmeier, H., Broer, I., 2006. Towards plant made vaccines. In: Plant Genetics - Joined Conference of the German Genetics Society and the German Society for Plant Breeding, Kiel/Germany, 20–23 September 2006, p. 139.
doi:10.1016/j.jbiotec.2007.07.072
Abstracts / Journal of Biotechnology 131S (2007) S43–S47
provides cross-protection against enterotoxic Escherichia coli. This vaccine is not available, however, in large part due to cost of production of the bacterial toxin protein which is a component of its formulation. The edible vaccines that at the moment are in clinical phases of research (Phase I) are the following ones: vaccine against the Enteritis caused by E. coli. expressed in potato plants; vaccine against infections caused by the virus Norwalk, expressed in potato plants and tobacco; vaccine against the virus of the Hepatitis B expressed in tobacco. Other fruits under experiment are banana and tomato. The latest research on topic are focused on multicomponent vaccines development that provide protection against several pathogens.
Mikschofsky, H., Schirrmeier, H., Broer, I., 2006. Towards plant made vaccines. In: Plant Genetics - Joined Conference of the German Genetics Society and the German Society for Plant Breeding, Kiel/Germany, 20–23 September 2006, p. 139.
doi:10.1016/j.jbiotec.2007.07.070
2 Friedrich-Loeffler-Institute
3. Plant made veterinary vaccines against RHD
The Rabbit hemorrhagic disease (RHD) is an acute and highly infectious viral disease affecting the European rabbit (Oryctolagus cuniculus). It causes high economic damages in rabbit meat production as well as limitations of wild populations. RHDvaccines are commercially produced by knowingly infected rabbits. VP60, the only structural capsid protein of RHDV, seems to be an appropriate subunit vaccine and offers possibilities to develop alternative vaccine production strategies. Plants demonstrate a promising production platform for vaccines, so far many different recombinant vaccines were expressed by higher plants -amongst others VP60 (Castanon et al., 2002; Gil et al., 2006). However, low expression levels combined with low immunogenicity of plant-derived VP60 prevented commercialisation of a plant-derived RHD-vaccine. Our aim was to enhance expression and immunogenicity of VP60 in different plant species in order to develop suitable plantderived RHD-vaccines. Different genetic modifications (codon adaptation, integration of regulatory and putative stabilizing sequences, and the addition of the well-known adjuvant ctb) led to a higher expression level as well to a tremendous enhancement of the immunogenicity of plant-derived VP60 (Mikschofsky et al., 2006). Surprisingly, only one of four synthetic ctbvp60 genes led to the expression of oligomers of pentameric CTB::VP60 in tobacco. This was only possible in tobacco and pea but not in potato, carrot or canola. Recombinant CTB::VP60 isolated from tobacco leaves demonstrated at least a 40fold higher immunogenicity compared to potato derived VP60-vaccines (Castanon et al., 2002; Castanon et al., 1999).
Heike Mikschofsky 1,∗ , Martin Hammer 2 , Patricia K¨onig 2 , G¨unther Keil 2 , Horst Schirrmeier 2 , Inge Broer 1 1 University
of Rostock, Rostock, Germany Insel, Riems, Germany
2 Friedrich-Loeffler-Institute
Vaccines against the rabbit hemorrhagic disease virus (RHDV), a highly infectious pathogen of the European rabbit (Oryctolagus cuniculus), are commercially produced by experimentally infected rabbits. VP60, the only structural capsid protein of RHDV, seems to be an appropriate subunit vaccine and offers possibilities to develop an alternative vaccine production strategy. Transgenic plants demonstrate a promising production platform for veterinary vaccines, up to now a number of viral and bacterial antigens were expressed by higher plants amongst others VP60 (Castanon et al., 2002; Gil et al., 2006). However, low expression levels and non-satisfying immunogenicity of plantderived VP60 prevented commercialisation of a plant-derived RHD-vaccine up to now. In order to develop suitable plant-derived RHD-vaccines we are studying the enhancement of the expression and the immunogenicity of VP60 in different plant species. Different genetic modifications (codon adaptation, integration of regulatory and putative stabilizing sequences, and the addition of the wellknown adjuvant ctb) led to a higher expression level as well as to a tremendous enhancement of the immunogenicity of plantderived VP60 (Mikschofsky et al., 2006). Interestingly, only one of four synthetic ctbvp60 genes led to the expression of oligomers of pentameric CTB::VP60. This was only possible in tobacco and pea but not in potato, carrot or canola. Tobacco and pea derived CTB::VP60 demonstrated at least a 40-fold higher immunogenicity compared to VP60-vaccines of potato tubers (Castanon et al., 1999, 2002). References Castanon, S., Martin-Alonso, J.M., Marin, M.S., Boga, J.A., Alonso, P., Parra, F., Ordas, R.J., 2002. Plant Sci. 162, 87–95. Castanon, S., Marin, M.S., Martin-Alonso, J.M., Boga, J.A., Casais, R., Humara, J.M., Ordas, R.J., Parra, F., 1999. J. Virol. 73, 4452–4455. Gil, F., Titarenko, E., Arcalis, E., Escribano, J.M., 2006. Plant Biotechnol. J. 4, 135–143.
doi:10.1016/j.jbiotec.2007.07.071 4. Production of high immunogenic VP60, a promising RHDvaccine, in plants Heike Mikschofsky 1,∗ , Martin Hammer 2 , Patricia K¨onig 2 , G¨unther Keil 2 , Horst Schirrmeier 2 , Inge Broer 1 1 University
of Rostock, Rostock, Germany Insel, Riems, Germany
References Castanon, S., Martin-Alonso, J.M., Marin, M.S., Boga, J.A., Alonso, P., Parra, F., Ordas, R.J., 2002. Plant Sci. 162, 87–95. Castanon, S., Marin, M.S., Martin-Alonso, J.M., Boga, J.A., Casais, R., Humara, J.M., Ordas, R.J., Parra, F., 1999. J. Virol. 73, 4452–4455. Gil, F., Titarenko, E., Arcalis, E., Escribano, J.M., 2006. Plant Biotechnol. J. 4, 135–143. Mikschofsky, H., Schirrmeier, H., Broer, I., 2006. Towards plant made vaccines. In: Plant Genetics - Joined Conference of the German Genetics Society and the German Society for Plant Breeding, Kiel/Germany, 20–23 September 2006, p. 139.
doi:10.1016/j.jbiotec.2007.07.072