CERELA   05438
CENTRO DE REFERENCIA PARA LACTOBACILOS
Unidad Ejecutora - UE
artículos
Título:
Production of Fibronectin Binding Protein A at the surface of Lactococcus lactis increases plasmid transfer in vitro and in vivo
Autor/es:
PONTES, D.; INNOVENTIN, S.; DEL CARMEN, S.; FRANCO ALMEIDA, J.; LEBLANC, J.G.; DE MORENO DE LEBLANC, A.; BLUGEON, S. ; CHERBUY, C.; LEFEVRE, D. ; AZEVEDO, V.; MIYOSHI, A.; LANGELLA, P.; CHATEL, J.M.
Revista:
PLOS ONE
Editorial:
PUBLIC LIBRARY SCIENCE
Referencias:
Lugar: San Francisco; Año: 2012 vol. 24 p. 1 - 1
ISSN:
1932-6203
Resumen:
Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNAdelivery vehicles. We previously showed that Lactococcus lactis (LL) expressing the Fibronectin-Binding Protein A ofStaphylococcus aureus (LL-FnBPA+) showed higher internalization rates in vitro in Caco-2 cells than the native (wt) lactococciand were able to deliver a eukaryotic Green Fluorescent Protein (GFP) expression plasmid in 1% of human Caco-2 cells. Here,using the bovine beta-lactoglobulin (BLG), one of the major cow?s milk allergen, and GFP we characterized the potential ofLL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying theplasmid pValac:BLG (LL-FnBPA+ BLG) was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Thenwe demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than theCaco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two differentmethods of detection, EIA and fluorescence microscopy, we showed in vivo that: i) in order to be effective, LL-FnBPA+required a pre-coating with Fetal Calf Serum before oral administration; ii) plasmid transfer occurred in enterocytes withoutregard to the strains used (invasive or not); iii) the use of LL-FnBPA+ increased the number of mice producing BLG, but notthe level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNAdelivery vector in vivo.