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 - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles Applied and Environmental Microbiology Year : 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

Abstract

Lactococci are noninvasive bacteria frequently used as protein delivery vectors and, more recently, as in vitro and in vivo DNA delivery vehicles. We previously showed that a functional eukaryotic enhanced green fluorescent protein (eGFP) expression plasmid vector was delivered in epithelial cells by Lactococcus lactis producing Listeria monocytogenes internalin A (L. lactis InlA_), but this strategy is limited in vivo to transgenic mice and guinea pigs. In this study, we compare the internalization ability of L. lactis InlA_ and L. lactis producing either the fibronectin-binding protein A of Staphylococcus aureus (L. lactis FnBPA_) or its fibronectin binding domains C and D (L. lactis CD_). L. lactis FnBPA_ and L. lactis InlA_ showed comparable internalization rates in Caco-2 cells, while the internalization rate observed with L. lactis CD_ was lower. As visualized by conventional and confocal fluorescence microscopy, large clusters of L. lactis FnBPA_, L. lactis CD_, and L. lactis InlA_ were present in the cytoplasm of Caco-2 cells after internalization. Moreover, the internalization rates of Lactobacillus acidophilus NCFM and of an NCFM mutant strain with the gene coding for the fibronectin- binding protein (fbpA) inactivated were also evaluated in Caco-2 cells. Similar low internalization rates were observed for both wild-type L. acidophilus NCFM and the fbpA mutant, suggesting that commensal fibronectin binding proteins have a role in adhesion but not in invasion. L. lactis FnBPA_, L. lactis CD_, and L. lactis InlA_ were then used to deliver a eukaryotic eGFP expression plasmid in Caco-2 cells: flow cytometry analysis showed that the highest percentage of green fluorescent Caco-2 cells was observed after coculture with either L. lactis FnBPA_ or L. lactis InlA_. Analysis of the in vivo efficiency of these invasive recombinant strains is currently in progress to validate their potential as DNA vaccine delivery vehicles.

Dates and versions

hal-02667480 , version 1 (31-05-2020)

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Silvia S. Innocentin, Valeria Guimaraes, Anderson Miyoshi, Vasco Azevedo, Philippe P. Langella, et al.. 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. Applied and Environmental Microbiology, 2009, 75 (14), pp.4870-4878. ⟨10.1128/AEM.00825-09⟩. ⟨hal-02667480⟩
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