Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum

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dc.contributor.authorKohl, Thomasen_ZA
dc.contributor.authorHitzeroth, Ingaen_ZA
dc.contributor.authorChristensen, Neilen_ZA
dc.contributor.authorRybicki, Edwarden_ZA
dc.date.accessioned2015-10-12T10:57:26Z
dc.date.available2015-10-12T10:57:26Z
dc.date.issued2007en_ZA
dc.description.abstractBACKGROUND:We have investigated the possibility and feasibility of producing the HPV-11 L1 major capsid protein in transgenic Arabidopsis thaliana ecotype Columbia and Nicotiana tabacum cv. Xanthi as potential sources for an inexpensive subunit vaccine. RESULTS: Transformation of plants was only achieved with the HPV-11 L1 gene with the C-terminal nuclear localization signal (NLS-) encoding region removed, and not with the full-length gene. The HPV-11 L1 NLS- gene was stably integrated and inherited through several generations of transgenic plants. Plant-derived HPV-11 L1 protein was capable of assembling into virus-like particles (VLPs), although resulting particles displayed a pleomorphic phenotype. Neutralising monoclonal antibodies binding both surface-linear and conformation-specific epitopes bound the A. thaliana-derived particles and - to a lesser degree - the N. tabacum-derived particles, suggesting that plant-derived and insect cell-derived VLPs displayed similar antigenic properties. Yields of up to 12 mug/g of HPV-11 L1 NLS- protein were harvested from transgenic A. thaliana plants, and 2 mug/g from N. tabacum plants - a significant increase over previous efforts. Immunization of New Zealand white rabbits with ~50 mug of plant-derived HPV-11 L1 NLS- protein induced an antibody response that predominantly recognized insect cell-produced HPV-11 L1 NLS- and not NLS+ VLPs. Evaluation of the same sera concluded that none of them were able to neutralise pseudovirion in vitro. CONCLUSION: We expressed the wild-type HPV-11 L1 NLS- gene in two different plant species and increased yields of HPV-11 L1 protein by between 500 and 1000-fold compared to previous reports. Inoculation of rabbits with extracts from both plant types resulted in a weak immune response, and antisera neither reacted with native HPV-11 L1 VLPs, nor did they neutralise HPV-11 pseudovirion infectivity. This has important and potentially negative implications for the production of HPV-11 vaccines in plants.en_ZA
dc.identifier.apacitationKohl, T., Hitzeroth, I., Christensen, N., & Rybicki, E. (2007). Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum. <i>BMC Biotechnology</i>, http://hdl.handle.net/11427/14191en_ZA
dc.identifier.chicagocitationKohl, Thomas, Inga Hitzeroth, Neil Christensen, and Edward Rybicki "Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum." <i>BMC Biotechnology</i> (2007) http://hdl.handle.net/11427/14191en_ZA
dc.identifier.citationKohl, T. O., Hitzeroth, I. I., Christensen, N. D., & Rybicki, E. P. (2007). Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum. BMC biotechnology, 7(1), 56.en_ZA
dc.identifier.ris TY - Journal Article AU - Kohl, Thomas AU - Hitzeroth, Inga AU - Christensen, Neil AU - Rybicki, Edward AB - BACKGROUND:We have investigated the possibility and feasibility of producing the HPV-11 L1 major capsid protein in transgenic Arabidopsis thaliana ecotype Columbia and Nicotiana tabacum cv. Xanthi as potential sources for an inexpensive subunit vaccine. RESULTS: Transformation of plants was only achieved with the HPV-11 L1 gene with the C-terminal nuclear localization signal (NLS-) encoding region removed, and not with the full-length gene. The HPV-11 L1 NLS- gene was stably integrated and inherited through several generations of transgenic plants. Plant-derived HPV-11 L1 protein was capable of assembling into virus-like particles (VLPs), although resulting particles displayed a pleomorphic phenotype. Neutralising monoclonal antibodies binding both surface-linear and conformation-specific epitopes bound the A. thaliana-derived particles and - to a lesser degree - the N. tabacum-derived particles, suggesting that plant-derived and insect cell-derived VLPs displayed similar antigenic properties. Yields of up to 12 mug/g of HPV-11 L1 NLS- protein were harvested from transgenic A. thaliana plants, and 2 mug/g from N. tabacum plants - a significant increase over previous efforts. Immunization of New Zealand white rabbits with ~50 mug of plant-derived HPV-11 L1 NLS- protein induced an antibody response that predominantly recognized insect cell-produced HPV-11 L1 NLS- and not NLS+ VLPs. Evaluation of the same sera concluded that none of them were able to neutralise pseudovirion in vitro. CONCLUSION: We expressed the wild-type HPV-11 L1 NLS- gene in two different plant species and increased yields of HPV-11 L1 protein by between 500 and 1000-fold compared to previous reports. Inoculation of rabbits with extracts from both plant types resulted in a weak immune response, and antisera neither reacted with native HPV-11 L1 VLPs, nor did they neutralise HPV-11 pseudovirion infectivity. This has important and potentially negative implications for the production of HPV-11 vaccines in plants. DA - 2007 DB - OpenUCT DO - 10.1186/1472-6750-7-56 DP - University of Cape Town J1 - BMC Biotechnology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2007 T1 - Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum TI - Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum UR - http://hdl.handle.net/11427/14191 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/14191
dc.identifier.urihttp://dx.doi.org/10.1186/1472-6750-7-56
dc.identifier.vancouvercitationKohl T, Hitzeroth I, Christensen N, Rybicki E. Expression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacum. BMC Biotechnology. 2007; http://hdl.handle.net/11427/14191.en_ZA
dc.language.isoengen_ZA
dc.publisherBioMed Central Ltden_ZA
dc.publisher.departmentInstitute of Infectious Disease and Molecular Medicineen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis is an Open Access article distributed under the terms of the Creative Commons Attribution Licenseen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/2.0en_ZA
dc.sourceBMC Biotechnologyen_ZA
dc.source.urihttp://www.biomedcentral.com/bmcbiotechnol/en_ZA
dc.subject.otherMolecular and Cell Biologyen_ZA
dc.titleExpression of HPV-11 L1 protein in transgenic Arabidopsis thaliana and Nicotiana tabacumen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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