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

BMC Biotechnology

Research Article

Malcolm J Fraser¹  Edward P Rybicki²  Fiona Tanzer²  Alisson G Lynch²  Enid G Shephard³  Anna-Lise Williamson⁴ 

[1] Department of Biological Sciences, University of Notre Dame, IN46556-0369, Notre Dame, USA;Department of Molecular and Cell Biology, Faculty of Science, University of Cape Town, Rondebosch, Cape Town, South Africa;Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, South Africa;Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, South Africa;MRC/UCT Liver Research Centre, Department of Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, South Africa;Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, South Africa;National Health Laboratory Service, Groote Schuur Hospital, Observatory, Cape Town, South Africa;

关键词: Insect Cell;    Inverted Terminal Repeat;    Insect Cell Line;    Rolling Circle Amplification;    Helper Plasmid;   

DOI  :  10.1186/1472-6750-10-30

 received in 2009-10-12, accepted in 2010-03-31,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundInsect 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.ResultsTransgenic 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.ConclusionTransgenic 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.

【 授权许可】

Unknown   
© Lynch et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.