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Applied and Environmental Microbiology, November 2006, p. 7091-7097, Vol. 72, No. 11
0099-2240/06/$08.00+0     doi:10.1128/AEM.01325-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Use of Native Lactococci as Vehicles for Delivery of DNA into Mammalian Epithelial Cells{triangledown}

Valéria Dellaretti Guimarães,1,2 Silvia Innocentin,1,3 François Lefèvre,1 Vasco Azevedo,2 Jean-Michel Wal,4 Philippe Langella,3 and Jean-Marc Chatel4*

Unité de Virologie et Immunologie Moléculaire, INRA, Domaine de Vilvert, 78352 Jouy en Josas Cedex, France,1 Institute of Biological Sciences, Federal University of Minas Gerais (UFMG-ICB), Belo Horizonte-MG, Brazil,2 Unité d'Ecologie et Physiologie du Système Digestif, INRA, Domaine de Vilvert, 78352 Jouy en Josas Cedex, France,3 Unité d'Immuno-Allergie Alimentaire, INRA-CEA, DRM-SPI, Bat 136, CEA de Saclay, 91191 Gif sur Yvette, France4

Received 9 June 2006/ Accepted 9 August 2006

The use of the food-grade bacterium Lactococcus lactis as a DNA delivery vehicle at the mucosal level is an attractive DNA vaccination strategy. Previous experiments showed that recombinant L. lactis expressing the Listeria monocytogenes inlA gene can deliver a functional gene into mammalian cells. Here, we explored the potential use of noninvasive L. lactis strains as a DNA delivery vehicle. We constructed two Escherichia coli-L. lactis shuttle plasmids, pLIG:BLG1 and pLIG:BLG2, containing a eukaryotic expression cassette with the cDNA of bovine ß-lactoglobulin (BLG). The greatest BLG expression after transfection of Cos-7 cells was obtained with pLIG:BLG1, which was then used to transform L. lactis MG1363. The resulting L. lactis strain MG1363(pLIG:BLG1) was not able to express BLG. The potential of L. lactis as a DNA delivery vehicle was analyzed by detection of BLG in Caco-2 human colon carcinoma cells after 3 h of coincubation with (i) purified pLIG:BLG1, (ii) MG1363(pLIG:BLG1), (iii) a mix of MG1363(pLIG) and purified pLIG:BLG1, and (iv) MG1363. Both BLG cDNA and BLG expression were detected only in Caco-2 cells coincubated with MG1363(pLIG:BLG1). There was a decrease in the BLG cDNA level in Caco-2 cells between 24 and 48 h after coincubation. BLG expression by Caco-2 cells started at 24 h and increased between 24 and 72 h. BLG secretion by Caco-2 cells started 48 h after coincubation with MG1363(pLIG:BLG1). We conclude that lactococci can deliver BLG cDNA into mammalian epithelial cells, demonstrating their potential to deliver in vivo a DNA vaccine.

* Corresponding author. Mailing address: Unité d'Immuno-Allergie Alimentaire, INRA-CEA, DRM-SPI, Bat. 136, CEA de Saclay, 91191 Gif sur Yvette, France. Phone: 33 1 69 08 80 04. Fax: 33 1 69 08 59 07. E-mail: jean-marc.chatel{at}cea.fr.

{triangledown} Published ahead of print on 8 September 2006.

Applied and Environmental Microbiology, November 2006, p. 7091-7097, Vol. 72, No. 11
0099-2240/06/$08.00+0     doi:10.1128/AEM.01325-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Innocentin, S., Guimaraes, V., Miyoshi, A., Azevedo, V., Langella, P., Chatel, J.-M., Lefevre, F. (2009). Lactococcus lactis Expressing either Staphylococcus aureus Fibronectin-Binding Protein A or Listeria monocytogenes Internalin A Can Efficiently Internalize and Deliver DNA in Human Epithelial Cells. Appl. Environ. Microbiol. 75: 4870-4878 [Abstract] [Full Text]  


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