Internalin-expressing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells

Guimarães, Valeria; Gabriel, Jane Eyre; Lefèvre, François; Cabanes, Didier; Gruss, Alexandra; Cossart, Pascale; Azevedo, Vasco; Langella, Philippe

doi:10.1016/j.micinf.2005.02.012

Cited by 69 publications

(78 citation statements)

References 43 publications

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“…21 After coculture of Caco-2 cells and L. lactis carrying a functional eukaryotic gfp gene, no fluorescent cells could be detected. 9 In similar experiments, the use of L. lactis LL(pLIG:BLG) to deliver a functional eukaryotic blg gene led to both expression and secretion of BLG. 3 In our experiments, the BLG cDNA was detected in DNA extracted from the small intestinal wall that was washed of gut contents 72 h after the last gavage.…”

Section: Discussionmentioning

confidence: 99%

“…7 Noninvasive bacteria as E. coli, L. lactis or Enterococcus faecalis were rendered invasive after expression of either inlA or inv gene. [8][9][10] Nevertheless, it has been demonstrated in vitro that noninvasive bacteria as L. lactis or E. coli are able to transfer genetic material. 3,9,11 In vivo, E. coli has been used as vector for DNA vaccines by intramuscular, intraperitoneal or subcutaneous administrations.…”

Section: Introductionmentioning

confidence: 99%

“…[8][9][10] Nevertheless, it has been demonstrated in vitro that noninvasive bacteria as L. lactis or E. coli are able to transfer genetic material. 3,9,11 In vivo, E. coli has been used as vector for DNA vaccines by intramuscular, intraperitoneal or subcutaneous administrations. 12,13 Here, we explored the ability of not genetically modified L. lactis to deliver plasmid in vivo to mice.…”

Section: Introductionmentioning

confidence: 99%

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In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells

Pothelune

Ah‐Leung

et al. 2008

Gene Ther

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We recently demonstrated that noninvasive food-grade Lactococcus lactis (L. lactis) can deliver eukaryotic expression plasmid in mammalian cells in vitro. Here, we evaluated, in vivo, whether a eukaryotic expression plasmid carried by lactococci can translocate to the epithelial cells of the intestinal membrane. The strain LL(pLIG:BLG1) carrying one plasmid containing a eukaryotic expression cassette encoding b-lactoglobulin (BLG), a major allergen of cow's milk, was orally administered by gavage to mice. BLG cDNA was detected in the epithelial membrane of the small intestine of 40% of the mice and BLG was produced in 53% of the mice. Oral administration of LL(pLIG:BLG1) induced a low and transitory Th1-type immune response counteracting a Th2 response in case of further sensitization. We demonstrated for the first time the transfer of a functional plasmid to the epithelial membrane of the small intestine in mice by noninvasive food-grade lactococci.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells

Pothelune

Ah‐Leung

et al. 2008

Gene Ther

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“…11,12) Those invasive L. lactis showed higher invasiveness to epithelial cells in vitro in comparison to the wild-type (wt) L. lactis, and resulted in the cellular expression of genes encoded by an eukaryotic expression plasmid.…”

Section: )mentioning

confidence: 99%

“…10) To improve the efficiency of L. lactis-mediated DNA delivery to intestinal cells, recombinant L. lactis strains expressing Internalin A (InlA), an invasin of intracellular bacteria L. monocytogenes, and those expressing fibronectin binding protein A (FnBPA) of Staphylococcus aureus were developed. 11,12) Those invasive L. lactis showed higher invasiveness to epithelial cells in vitro in comparison to the wild-type (wt) L. lactis, and resulted in the cellular expression of genes encoded by an eukaryotic expression plasmid. 11,12) The interaction of InlA with E-cadherin induces the endocytosis of InlA-expressing bacteria.…”

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confidence: 99%

Peyer’s Patches as a Portal for DNA Delivery by <i>Lactococcus lactis</i> <i>in Vivo</i>

Yano

Takahashi

Mori

et al. 2018

Biological & Pharmaceutical Bulletin

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Application of food-grade Lactococcus lactis (L. lactis) as a safe delivery tool for DNA vaccines and therapeutic proteins has been well investigated. Although some studies showed that eukaryotic expression plasmids were transferred from L. lactis to enterocytes, the precise mechanism of the DNA transfer remains unknown. In this study, we generated an invasive L. lactis strain that expresses "murinized" Internalin A, an invasin of intracellular bacteria Listeria monocytogenes with two amino acid alterations for invasion into murine cells, and confirmed that this L. lactis strain delivered DNA in an invasin-dependent manner into a monolayer of epithelial cells polarized to mimic the gastrointestinal tract environment. Although invasive L. lactis inoculated orally can deliver DNA into enterocytes in the gastrointestinal tract of mice, the efficiency of DNA transfer was similar to that of non-invasive L. lactis strain, suggesting that the in vivo DNA transfer from L. lactis occurs invasin-independently. A ligated-intestinal loop assay, a method for a short-term culturing of the whole intestine filled with materials to evaluate the interaction of the materials with intestinal cells, demonstrated that both non-invasive and invasive L. lactis strains were present in the Peyer's patches of the small intestine. On the other hand, few L. lactis was detected in the non-Peyer's patch epithelial region. Thus, our observations lead us to speculate that DNA transfer from L. lactis occurs predominantly in the Peyer's patches in an invasin-independent manner. Key words Lactococcus lactis;Peyer's patch; DNA delivery; Internalin A Vaccination with a plasmid containing the DNA sequence encoding an antigen has been of great interest of late as it can induce both humoral and cell-mediated immune responses. 1)Despite numerous studies, no DNA vaccines have been licensed for human use to date. The administration of naked DNA is less immunogenic in large animals and humans, probably due to the low level of protein expression.2) To improve the potency of DNA vaccination, various DNA delivery platforms, including electroporation, needle-free jet injection and liposomes, have been developed. 3,4) The use of bacteria as a delivery vehicle for DNA vaccines has emerged recently as a promising approach to enhance potency, 5) and one of the attractive features of the use of bacteria is the potential for oral administration to induce not only systemic but also mucosal immune responses. 5) Salmonella typhimurium and Listeria monocytogenes (L. monocytogenes) have been studied as experimental delivery vehicles as they are known to invade intestinal epithelial cells. 6) Even though the strains employed are extensively attenuated for virulence, the possibility of reversion to virulence as well as preexisting immunity limits their clinical use, especially for infants, the elderly and immunocompromised hosts. 7,8) The mechanisms involving DNA transfer from bacteria to mammalian cells and tissues remain to be clarified.The use of food-grade Lactococcus ...

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Gastrointestinal Disorders: A Patho‐biotechnology Approach to Probiotic Therapy

Sleator¹

2014

In Vivo Models for Drug Discovery

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Internalin-expressing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells

Cited by 69 publications

References 43 publications

In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells

In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells

Peyer’s Patches as a Portal for DNA Delivery by <i>Lactococcus lactis</i> <i>in Vivo</i>

Gastrointestinal Disorders: A Patho‐biotechnology Approach to Probiotic Therapy

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