A new plasmid vector for DNA delivery using lactococci

doi:10.1186/1479-0556-7-4

Genetic Vaccines and Therapy

Volume 7

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Guimarães V
Innocentin S
Chatel JM
Lefèvre F
Langella P
Azevedo V
Miyoshi A

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Innocentin S
Chatel JM
Lefèvre F
Langella P
Azevedo V
Miyoshi A
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Research

A new plasmid vector for DNA delivery using lactococci

Valeria Guimarães¹ , Sylvia Innocentin² , Jean-Marc Chatel³ , François Lefèvre⁴ , Philippe Langella² , Vasco Azevedo¹ and Anderson Miyoshi¹

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

⁴INRA, UR892, Unité de Virologie et Immunologie Moléculaires, Domaine de Vilvert, 78352 Jouy-en-Josas, France

author email corresponding author email

Genetic Vaccines and Therapy 2009, 7:4doi:10.1186/1479-0556-7-4


Published:	10 February 2009

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.