Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Simple item page
dc.contributor.authorLynch, Alissonen_ZA
dc.contributor.authorTanzer, Fionaen_ZA
dc.contributor.authorFraser, Malcolmen_ZA
dc.contributor.authorShephard, Eniden_ZA
dc.contributor.authorWilliamson, Anna-Liseen_ZA
dc.contributor.authorRybicki, Edwarden_ZA
dc.date.accessioned2015-11-11T12:00:06Z
dc.date.available2015-11-11T12:00:06Z
dc.date.issued2010en_ZA
dc.description.abstractBACKGROUND: Insect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system. RESULTS: Transgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs. CONCLUSION: Transgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron.en_ZA
dc.identifier.apacitationLynch, A., Tanzer, F., Fraser, M., Shephard, E., Williamson, A., & Rybicki, E. (2010). Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production. <i>BMC Biotechnology</i>, http://hdl.handle.net/11427/14882en_ZA
dc.identifier.chicagocitationLynch, Alisson, Fiona Tanzer, Malcolm Fraser, Enid Shephard, Anna-Lise Williamson, and Edward Rybicki "Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production." <i>BMC Biotechnology</i> (2010) http://hdl.handle.net/11427/14882en_ZA
dc.identifier.citationLynch, A. G., Tanzer, F., Fraser, M. J., Shephard, E. G., Williamson, A. L., & Rybicki, E. P. (2010). Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production. BMC biotechnology, 10(1), 30.en_ZA
dc.identifier.ris TY - Journal Article AU - Lynch, Alisson AU - Tanzer, Fiona AU - Fraser, Malcolm AU - Shephard, Enid AU - Williamson, Anna-Lise AU - Rybicki, Edward AB - BACKGROUND: Insect baculovirus-produced Human immunodeficiency virus type 1 (HIV-1) Gag virus-like-particles (VLPs) stimulate good humoral and cell-mediated immune responses in animals and are thought to be suitable as a vaccine candidate. Drawbacks to this production system include contamination of VLP preparations with baculovirus and the necessity for routine maintenance of infectious baculovirus stock. We used piggyBac transposition as a novel method to create transgenic insect cell lines for continuous VLP production as an alternative to the baculovirus system. RESULTS: Transgenic cell lines maintained stable gag transgene integration and expression up to 100 cell passages, and although the level of VLPs produced was low compared to baculovirus-produced VLPs, they appeared similar in size and morphology to baculovirus-expressed VLPs. In a murine immunogenicity study, whereas baculovirus-produced VLPs elicited good CD4 immune responses in mice when used to boost a prime with a DNA vaccine, no boost response was elicited by transgenically produced VLPs. CONCLUSION: Transgenic insect cells are stable and can produce HIV Pr55 Gag VLPs for over 100 passages: this novel result may simplify strategies aimed at making protein subunit vaccines for HIV. Immunogenicity of the Gag VLPs in mice was less than that of baculovirus-produced VLPs, which may be due to lack of baculovirus glycoprotein incorporation in the transgenic cell VLPs. Improved yield and immunogenicity of transgenic cell-produced VLPs may be achieved with the addition of further genetic elements into the piggyBac integron. DA - 2010 DB - OpenUCT DO - 10.1186/1472-6750-10-30 DP - University of Cape Town J1 - BMC Biotechnology LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 T1 - Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production TI - Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production UR - http://hdl.handle.net/11427/14882 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/14882
dc.identifier.urihttp://dx.doi.org/10.1186/1472-6750-10-30
dc.identifier.vancouvercitationLynch A, Tanzer F, Fraser M, Shephard E, Williamson A, Rybicki E. Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production. BMC Biotechnology. 2010; http://hdl.handle.net/11427/14882.en_ZA
dc.language.isoengen_ZA
dc.publisherBioMed Central Ltden_ZA
dc.publisher.departmentDepartment of Molecular and Cell Biologyen_ZA
dc.publisher.facultyFaculty of Scienceen_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.holder2010 Lynch et al; licensee BioMed Central Ltd.en_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.otherAIDS Vaccinesen_ZA
dc.subject.otherBaculoviridaeen_ZA
dc.subject.otherHIV-1en_ZA
dc.titleUse of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP productionen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Lynch_Use_of_the_piggyBac_transposon_2010.pdf
Size:
1.44 MB
Format:
Adobe Portable Document Format
Description:
Download
Collections
Journal Articles