A new plasmid vector for DNA delivery using lactococci
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BioMed Central
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A new plasmid vector for DNA delivery using lactococci Valeria Guimarães1, Sylvia Innocentin2, Jean-Marc Chatel3, François Lefèvre4, Philippe Langella2, Vasco Azevedo1 and Anderson Miyoshi*1 Address: 1Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB-UFMG), Belo Horizonte – MG, Brasil, 2INRA, UR910, Unité d'Ecologie et Physiologie du Système Digestif, Domaine de Vilvert, 78352 Jouy-en-Josas, France, 3INRA, UR496, Unité d'Immuno-Allergie Alimentaire, Domaine de Vilvert, 78352 Jouy-en-Josas, France and 4INRA, UR892, Unité de Virologie et Immunologie Moléculaires, Domaine de Vilvert, 78352 Jouy-en-Josas, France Email: Valeria Guimarães - [email protected]; Sylvia Innocentin - [email protected]; JeanMarc Chatel - [email protected]; François Lefèvre - [email protected]; Philippe Langella - [email protected]; Vasco Azevedo - [email protected]; Anderson Miyoshi* - [email protected] * Corresponding author
Published: 10 February 2009 Genetic Vaccines and Therapy 2009, 7:4
doi:10.1186/1479-0556-7-4
Received: 15 October 2008 Accepted: 10 February 2009
This article is available from: http://www.gvt-journal.com/content/7/1/4 © 2009 Guimarães et al; licensee 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.
Abstract Background: The use of food-grade lactococci as bacterial carriers to DNA delivery into epithelial cells is a new strategy to develop live oral DNA vaccine. Our goal was to develop a new plasmid, named pValac, for antigen delivery for use in lactococci. The pValac plasmid was constructed by the fusion of: i) a eukaryotic region, allowing the cloning of an antigen of interest under the control of the pCMV eukaryotic promoter to be expressed by a host cell and ii) a prokaryotic region allowing replication and selection of bacteria. In order to evaluate pValac functionality, the gfp ORF was cloned into pValac (pValac:gfp) and was analysed by transfection in PK15 cells. The applicability of pValac was demonstrated by invasiveness assays of Lactococcus lactis inlA+ strains harbouring pValac:gfp into Caco-2 cells. Results: After transfection with pValac:gfp, we observed GFP expression in PK15 cells. L. lactis inlA+ were able to invade Caco-2 cells and delivered a functional expression cassette (pCMV:gfp) into epithelial cells. Conclusion: We showed the potential of an invasive L. lactis harbouring pValac to DNA delivery and subsequent triggering DNA expression by epithelial cells. Further work will be to examine whether these strains are able to deliver DNA in intestinal cells in vivo.
Background Numerous infectious agents invade the host through the mucosa to cause disease. The use of bacterial carriers to deliver DNA vaccine by oral route constitutes a promising
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