A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants

The expression of antigens in transgenic plants has been increasingly used as an alternative to the classical methodologies for antigen expression in the development of experimental vaccines. However, an important limitation in most cases is the low concentration of the recombinant antigens in the p...

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Autores principales:	Dus Santos, M.J., Wigdorovitz, A., Trono, K., Ríos, R.D., Franzone, P.M., Gil, F., Moreno, J., Carrillo, C., Escribano, J.M., Borca, M.V.
Formato:	JOUR
Materias:	Antigen expression FMDV Recombinant plants Vaccination amino acid beta glucuronidase epitope foot and mouth disease vaccine peptide protein VP1 alfalfa analytic method animal experiment animal model antibody response antigen expression article controlled study enzyme activity experimental host foot and mouth disease Foot and mouth disease virus gene insertion immune response immunization immunogenicity methodology mouse nonhuman priority journal protection protein expression provocation test reporter gene transgenic plant vaccine production virus particle virus virulence Animals Antibodies, Viral Capsid Capsid Proteins Foot-and-Mouth Disease Virus Immunization Mice Peptide Fragments Plants, Genetically Modified Vaccines, Synthetic Viral Vaccines
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_0264410X_v20_n7-8_p1141_DusSantos
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_0264410X_v20_n7-8_p1141_DusSantos2023-10-03T15:12:48Z A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants Dus Santos, M.J. Wigdorovitz, A. Trono, K. Ríos, R.D. Franzone, P.M. Gil, F. Moreno, J. Carrillo, C. Escribano, J.M. Borca, M.V. Antigen expression FMDV Recombinant plants Vaccination amino acid beta glucuronidase epitope foot and mouth disease vaccine peptide protein VP1 alfalfa analytic method animal experiment animal model antibody response antigen expression article controlled study enzyme activity experimental host foot and mouth disease Foot and mouth disease virus gene insertion immune response immunization immunogenicity methodology mouse nonhuman priority journal protection protein expression provocation test reporter gene transgenic plant vaccine production virus particle virus virulence Animals Antibodies, Viral Capsid Capsid Proteins Foot-and-Mouth Disease Virus Immunization Mice Peptide Fragments Plants, Genetically Modified Vaccines, Synthetic Viral Vaccines The expression of antigens in transgenic plants has been increasingly used as an alternative to the classical methodologies for antigen expression in the development of experimental vaccines. However, an important limitation in most cases is the low concentration of the recombinant antigens in the plant tissues, which reduces the possibilities of practical applications. Because the site of insertion of the transferred DNA into the cellular chromosomal DNA is at random, different levels of foreign protein expression in independent transformants is expected. Strategies to allow the evaluation of a high number of the transgenic individuals, usually an expensive and very time consuming process, would permit the selection of those plants presenting the highest levels of recombinant protein expression. Here, we present the development of an experimental immunogen based in the expression of a highly immunogenic epitope from foot and mouth disease virus (FMDV) fused to the glucuronidase (gus A) reporter gene, which allows selection of the transgenic plants by the ß-glucuronidase (ßGUS) enzymatic activity. We produced transgenic plants of alfalfa expressing the immunogenic site between amino acid residues 135-160 of structural protein VP1 (VP135-160), fused to the ßGUS protein. Plants expressing the highest levels of the immunogenic epitope VP135-160, analyzed by Western blot, were efficiently selected based on their levels of ßGUS enzymatic activity. The FMDV epitope expressed in plants was highly immunogenic in mice which developed, after immunization, a strong anti-FMDV antibody response against a synthetic peptide representing the region VP135-160, to native virus VP1, and to purified FMDV particles. Additionally, these mice were completely protected against experimental challenge with the virulent virus. To our knowledge, this constitutes the first report of a peptide-based vaccine produced in transgenic plants that induces a protective immune response when used in experimental hosts. Also, these results demonstrated the possibility of using a novel and simple methodology for obtaining transgenic plants expressing high levels of foreign immunogenic epitopes, which could be directly applied in the development of plant-based vaccines. © 2002 Elsevier Science Ltd. All rights reserved. Fil:Dus Santos, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Wigdorovitz, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Moreno, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0264410X_v20_n7-8_p1141_DusSantos
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Antigen expression FMDV Recombinant plants Vaccination amino acid beta glucuronidase epitope foot and mouth disease vaccine peptide protein VP1 alfalfa analytic method animal experiment animal model antibody response antigen expression article controlled study enzyme activity experimental host foot and mouth disease Foot and mouth disease virus gene insertion immune response immunization immunogenicity methodology mouse nonhuman priority journal protection protein expression provocation test reporter gene transgenic plant vaccine production virus particle virus virulence Animals Antibodies, Viral Capsid Capsid Proteins Foot-and-Mouth Disease Virus Immunization Mice Peptide Fragments Plants, Genetically Modified Vaccines, Synthetic Viral Vaccines
spellingShingle	Antigen expression FMDV Recombinant plants Vaccination amino acid beta glucuronidase epitope foot and mouth disease vaccine peptide protein VP1 alfalfa analytic method animal experiment animal model antibody response antigen expression article controlled study enzyme activity experimental host foot and mouth disease Foot and mouth disease virus gene insertion immune response immunization immunogenicity methodology mouse nonhuman priority journal protection protein expression provocation test reporter gene transgenic plant vaccine production virus particle virus virulence Animals Antibodies, Viral Capsid Capsid Proteins Foot-and-Mouth Disease Virus Immunization Mice Peptide Fragments Plants, Genetically Modified Vaccines, Synthetic Viral Vaccines Dus Santos, M.J. Wigdorovitz, A. Trono, K. Ríos, R.D. Franzone, P.M. Gil, F. Moreno, J. Carrillo, C. Escribano, J.M. Borca, M.V. A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
topic_facet	Antigen expression FMDV Recombinant plants Vaccination amino acid beta glucuronidase epitope foot and mouth disease vaccine peptide protein VP1 alfalfa analytic method animal experiment animal model antibody response antigen expression article controlled study enzyme activity experimental host foot and mouth disease Foot and mouth disease virus gene insertion immune response immunization immunogenicity methodology mouse nonhuman priority journal protection protein expression provocation test reporter gene transgenic plant vaccine production virus particle virus virulence Animals Antibodies, Viral Capsid Capsid Proteins Foot-and-Mouth Disease Virus Immunization Mice Peptide Fragments Plants, Genetically Modified Vaccines, Synthetic Viral Vaccines
description	The expression of antigens in transgenic plants has been increasingly used as an alternative to the classical methodologies for antigen expression in the development of experimental vaccines. However, an important limitation in most cases is the low concentration of the recombinant antigens in the plant tissues, which reduces the possibilities of practical applications. Because the site of insertion of the transferred DNA into the cellular chromosomal DNA is at random, different levels of foreign protein expression in independent transformants is expected. Strategies to allow the evaluation of a high number of the transgenic individuals, usually an expensive and very time consuming process, would permit the selection of those plants presenting the highest levels of recombinant protein expression. Here, we present the development of an experimental immunogen based in the expression of a highly immunogenic epitope from foot and mouth disease virus (FMDV) fused to the glucuronidase (gus A) reporter gene, which allows selection of the transgenic plants by the ß-glucuronidase (ßGUS) enzymatic activity. We produced transgenic plants of alfalfa expressing the immunogenic site between amino acid residues 135-160 of structural protein VP1 (VP135-160), fused to the ßGUS protein. Plants expressing the highest levels of the immunogenic epitope VP135-160, analyzed by Western blot, were efficiently selected based on their levels of ßGUS enzymatic activity. The FMDV epitope expressed in plants was highly immunogenic in mice which developed, after immunization, a strong anti-FMDV antibody response against a synthetic peptide representing the region VP135-160, to native virus VP1, and to purified FMDV particles. Additionally, these mice were completely protected against experimental challenge with the virulent virus. To our knowledge, this constitutes the first report of a peptide-based vaccine produced in transgenic plants that induces a protective immune response when used in experimental hosts. Also, these results demonstrated the possibility of using a novel and simple methodology for obtaining transgenic plants expressing high levels of foreign immunogenic epitopes, which could be directly applied in the development of plant-based vaccines. © 2002 Elsevier Science Ltd. All rights reserved.
format	JOUR
author	Dus Santos, M.J. Wigdorovitz, A. Trono, K. Ríos, R.D. Franzone, P.M. Gil, F. Moreno, J. Carrillo, C. Escribano, J.M. Borca, M.V.
author_facet	Dus Santos, M.J. Wigdorovitz, A. Trono, K. Ríos, R.D. Franzone, P.M. Gil, F. Moreno, J. Carrillo, C. Escribano, J.M. Borca, M.V.
author_sort	Dus Santos, M.J.
title	A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
title_short	A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
title_full	A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
title_fullStr	A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
title_full_unstemmed	A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants
title_sort	novel methodology to develop a foot and mouth disease virus (fmdv) peptide-based vaccine in transgenic plants
url	http://hdl.handle.net/20.500.12110/paper_0264410X_v20_n7-8_p1141_DusSantos
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A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants

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