Expression of HIV-1 antigens in plants as potential subunit vaccines

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dc.contributor.authorMeyers, Annen_ZA
dc.contributor.authorChakauya, Erecken_ZA
dc.contributor.authorShephard, Eniden_ZA
dc.contributor.authorTanzer, Fionaen_ZA
dc.contributor.authorMaclean, Jamesen_ZA
dc.contributor.authorLynch, Alissonen_ZA
dc.contributor.authorWilliamson, Anna-Liseen_ZA
dc.contributor.authorRybicki, Edwarden_ZA
dc.date.accessioned2015-10-28T07:01:14Z
dc.date.available2015-10-28T07:01:14Z
dc.date.issued2008en_ZA
dc.description.abstractBACKGROUND: Human immunodeficiency virus type 1 (HIV-1) has infected more than 40 million people worldwide, mainly in sub-Saharan Africa. The high prevalence of HIV-1 subtype C in southern Africa necessitates the development of cheap, effective vaccines. One means of production is the use of plants, for which a number of different techniques have been successfully developed. HIV-1 Pr55Gag is a promising HIV-1 vaccine candidate: we compared the expression of this and a truncated Gag (p17/p24) and the p24 capsid subunit in Nicotiana spp. using transgenic plants and transient expression via Agrobacterium tumefaciens and recombinant tobamovirus vectors. We also investigated the influence of subcellular localisation of recombinant protein to the chloroplast and the endoplasmic reticulum (ER) on protein yield. We partially purified a selected vaccine candidate and tested its stimulation of a humoral and cellular immune response in mice. RESULTS: Both transient and transgenic expression of the HIV antigens were successful, although expression of Pr55Gag was low in all systems; however, the Agrobacterium-mediated transient expression of p24 and p17/p24 yielded best, to more than 1 mg p24/kg fresh weight. Chloroplast targeted protein levels were highest in transient and transgenic expression of p24 and p17/p24. The transiently-expressed p17/p24 was not immunogenic in mice as a homologous vaccine, but it significantly boosted a humoral and T cell immune response primed by a gag DNA vaccine, pTHGagC. CONCLUSION: Transient agroinfiltration was best for expression of all of the recombinant proteins tested, and p24 and p17/p24 were expressed at much higher levels than Pr55Gag. Our results highlight the usefulness of plastid signal peptides in enhancing the production of recombinant proteins meant for use as vaccines. The p17/p24 protein effectively boosted T cell and humoral responses in mice primed by the DNA vaccine pTHGagC, showing that this plant-produced protein has potential for use as a vaccine.en_ZA
dc.identifier.apacitationMeyers, A., Chakauya, E., Shephard, E., Tanzer, F., Maclean, J., Lynch, A., ... Rybicki, E. (2008). Expression of HIV-1 antigens in plants as potential subunit vaccines. <i>BMC Biotechnology</i>, http://hdl.handle.net/11427/14452en_ZA
dc.identifier.chicagocitationMeyers, Ann, Ereck Chakauya, Enid Shephard, Fiona Tanzer, James Maclean, Alisson Lynch, Anna-Lise Williamson, and Edward Rybicki "Expression of HIV-1 antigens in plants as potential subunit vaccines." <i>BMC Biotechnology</i> (2008) http://hdl.handle.net/11427/14452en_ZA
dc.identifier.citationMeyers, A., Chakauya, E., Shephard, E., Tanzer, F. L., Maclean, J., Lynch, A., ... & Rybicki, E. P. (2008). Expression of HIV-1 antigens in plants as potential subunit vaccines. BMC biotechnology, 8(1), 53.en_ZA
dc.identifier.ris TY - Journal Article AU - Meyers, Ann AU - Chakauya, Ereck AU - Shephard, Enid AU - Tanzer, Fiona AU - Maclean, James AU - Lynch, Alisson AU - Williamson, Anna-Lise AU - Rybicki, Edward AB - BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) has infected more than 40 million people worldwide, mainly in sub-Saharan Africa. The high prevalence of HIV-1 subtype C in southern Africa necessitates the development of cheap, effective vaccines. One means of production is the use of plants, for which a number of different techniques have been successfully developed. HIV-1 Pr55Gag is a promising HIV-1 vaccine candidate: we compared the expression of this and a truncated Gag (p17/p24) and the p24 capsid subunit in Nicotiana spp. using transgenic plants and transient expression via Agrobacterium tumefaciens and recombinant tobamovirus vectors. We also investigated the influence of subcellular localisation of recombinant protein to the chloroplast and the endoplasmic reticulum (ER) on protein yield. We partially purified a selected vaccine candidate and tested its stimulation of a humoral and cellular immune response in mice. RESULTS: Both transient and transgenic expression of the HIV antigens were successful, although expression of Pr55Gag was low in all systems; however, the Agrobacterium-mediated transient expression of p24 and p17/p24 yielded best, to more than 1 mg p24/kg fresh weight. Chloroplast targeted protein levels were highest in transient and transgenic expression of p24 and p17/p24. The transiently-expressed p17/p24 was not immunogenic in mice as a homologous vaccine, but it significantly boosted a humoral and T cell immune response primed by a gag DNA vaccine, pTHGagC. CONCLUSION: Transient agroinfiltration was best for expression of all of the recombinant proteins tested, and p24 and p17/p24 were expressed at much higher levels than Pr55Gag. Our results highlight the usefulness of plastid signal peptides in enhancing the production of recombinant proteins meant for use as vaccines. The p17/p24 protein effectively boosted T cell and humoral responses in mice primed by the DNA vaccine pTHGagC, showing that this plant-produced protein has potential for use as a vaccine. DA - 2008 DB - OpenUCT DO - 10.1186/1472-6750-8-53 DP - University of Cape Town J1 - BMC Biotechnology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2008 T1 - Expression of HIV-1 antigens in plants as potential subunit vaccines TI - Expression of HIV-1 antigens in plants as potential subunit vaccines UR - http://hdl.handle.net/11427/14452 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/14452
dc.identifier.urihttp://dx.doi.org/10.1186/1472-6750-8-53
dc.identifier.vancouvercitationMeyers A, Chakauya E, Shephard E, Tanzer F, Maclean J, Lynch A, et al. Expression of HIV-1 antigens in plants as potential subunit vaccines. BMC Biotechnology. 2008; http://hdl.handle.net/11427/14452.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.holder2008 Meyers et al; licensee BioMed Central Ltden_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/2.0/en_ZA
dc.sourceBMC Biotechnologyen_ZA
dc.source.urihttp://www.biomedcentral.com/bmcbiotechnol/en_ZA
dc.subject.otherAgrobacterium tumefaciensen_ZA
dc.subject.otherAIDS Vaccinesen_ZA
dc.subject.otherHIV Antigensen_ZA
dc.subject.otherTobaccoen_ZA
dc.subject.otherPlants, Genetically Modifieden_ZA
dc.titleExpression of HIV-1 antigens in plants as potential subunit vaccinesen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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