Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo

Autores
Pontes, Daniela Santos; Innocentin, Silvia; del Carmen, Silvina Andrea; Franco Almeida, Juliana; Leblanc, Jean Guy Joseph; de Moreno, Maria Alejandra; Blugeon, Sébastien; Cherbuy, Claire; Lefèvre, François; Azevedo, Vasco; Miyoshi, Anderson; Langella, Philippe; Chatel, Jean-Marc
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNA delivery vehicles. We previously showed that Lactococcus lactis (LL) expressing the Fibronectin-Binding Protein A of Staphylococcus aureus (LL-FnBPA+) showed higher internalization rates in vitro in Caco-2 cells than the native (wt) lactococci and 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 of LL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying the plasmid pValac:BLG (LL-FnBPA+ BLG) was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Then we demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than the Caco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two different methods 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 without regard to the strains used (invasive or not); iii) the use of LL-FnBPA+ increased the number of mice producing BLG, but not the level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNA delivery vector in vivo.
Fil: Pontes, Daniela Santos. Institut National de la Recherche Agronomique; Francia
Fil: Innocentin, Silvia. Institut National de la Recherche Agronomique; Francia
Fil: del Carmen, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Franco Almeida, Juliana. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: de Moreno, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Blugeon, Sébastien. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Cherbuy, Claire. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Lefèvre, François. Institut National de la Recherche Agronomique; Francia
Fil: Azevedo, Vasco. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; Brasil
Fil: Miyoshi, Anderson. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; Brasil
Fil: Langella, Philippe. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Chatel, Jean-Marc. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Materia
FIBRONECTIN-BINDING PROTEIN A
LACTOCOCCUS LACTIS
BOVINE BETA-LACTOGLOBULIN
DNA DELIVERY
Nivel de accesibilidad
acceso abierto
Condiciones de uso
Atribución-NoComercial-CompartirIgual 2.5 Argentina (CC BY-NC-SA 2.5 AR)
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/25289
Acceder
id CONICETDig_e0dd7de9f0cd59628f86825a93d819ab
oai_identifier_str oai:ri.conicet.gov.ar:11336/25289
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivoPontes, Daniela SantosInnocentin, Silviadel Carmen, Silvina AndreaFranco Almeida, JulianaLeblanc, Jean Guy Josephde Moreno, Maria AlejandraBlugeon, SébastienCherbuy, ClaireLefèvre, FrançoisAzevedo, VascoMiyoshi, AndersonLangella, PhilippeChatel, Jean-MarcFIBRONECTIN-BINDING PROTEIN ALACTOCOCCUS LACTISBOVINE BETA-LACTOGLOBULINDNA DELIVERYhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNA delivery vehicles. We previously showed that Lactococcus lactis (LL) expressing the Fibronectin-Binding Protein A of Staphylococcus aureus (LL-FnBPA+) showed higher internalization rates in vitro in Caco-2 cells than the native (wt) lactococci and 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 of LL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying the plasmid pValac:BLG (LL-FnBPA+ BLG) was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Then we demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than the Caco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two different methods 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 without regard to the strains used (invasive or not); iii) the use of LL-FnBPA+ increased the number of mice producing BLG, but not the level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNA delivery vector in vivo.Fil: Pontes, Daniela Santos. Institut National de la Recherche Agronomique; FranciaFil: Innocentin, Silvia. Institut National de la Recherche Agronomique; FranciaFil: del Carmen, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Franco Almeida, Juliana. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; FranciaFil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: de Moreno, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Blugeon, Sébastien. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; FranciaFil: Cherbuy, Claire. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; FranciaFil: Lefèvre, François. Institut National de la Recherche Agronomique; FranciaFil: Azevedo, Vasco. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; BrasilFil: Miyoshi, Anderson. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; BrasilFil: Langella, Philippe. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; FranciaFil: Chatel, Jean-Marc. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; FranciaPublic Library of Science2012-09-27info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/25289Pontes, Daniela Santos; Innocentin, Silvia; del Carmen, Silvina Andrea; Franco Almeida, Juliana; Leblanc, Jean Guy Joseph; et al.; Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo; Public Library of Science; Plos One; 7; 9; 27-9-2012; 1-6; e44892-e448921932-62031932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0044892info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0044892info:eu-repo/semantics/openAccessAtribución-NoComercial-CompartirIgual 2.5 Argentina (CC BY-NC-SA 2.5 AR)https://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:53:19Zoai:ri.conicet.gov.ar:11336/25289instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:53:20.057CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
title Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
spellingShingle Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
Pontes, Daniela Santos
FIBRONECTIN-BINDING PROTEIN A
LACTOCOCCUS LACTIS
BOVINE BETA-LACTOGLOBULIN
DNA DELIVERY
title_short Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
title_full Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
title_fullStr Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
title_full_unstemmed Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
title_sort Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo
dc.creator.none.fl_str_mv Pontes, Daniela Santos
Innocentin, Silvia
del Carmen, Silvina Andrea
Franco Almeida, Juliana
Leblanc, Jean Guy Joseph
de Moreno, Maria Alejandra
Blugeon, Sébastien
Cherbuy, Claire
Lefèvre, François
Azevedo, Vasco
Miyoshi, Anderson
Langella, Philippe
Chatel, Jean-Marc
author Pontes, Daniela Santos
author_facet Pontes, Daniela Santos
Innocentin, Silvia
del Carmen, Silvina Andrea
Franco Almeida, Juliana
Leblanc, Jean Guy Joseph
de Moreno, Maria Alejandra
Blugeon, Sébastien
Cherbuy, Claire
Lefèvre, François
Azevedo, Vasco
Miyoshi, Anderson
Langella, Philippe
Chatel, Jean-Marc
author_role author
author2 Innocentin, Silvia
del Carmen, Silvina Andrea
Franco Almeida, Juliana
Leblanc, Jean Guy Joseph
de Moreno, Maria Alejandra
Blugeon, Sébastien
Cherbuy, Claire
Lefèvre, François
Azevedo, Vasco
Miyoshi, Anderson
Langella, Philippe
Chatel, Jean-Marc
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv FIBRONECTIN-BINDING PROTEIN A
LACTOCOCCUS LACTIS
BOVINE BETA-LACTOGLOBULIN
DNA DELIVERY
topic FIBRONECTIN-BINDING PROTEIN A
LACTOCOCCUS LACTIS
BOVINE BETA-LACTOGLOBULIN
DNA DELIVERY
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNA delivery vehicles. We previously showed that Lactococcus lactis (LL) expressing the Fibronectin-Binding Protein A of Staphylococcus aureus (LL-FnBPA+) showed higher internalization rates in vitro in Caco-2 cells than the native (wt) lactococci and 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 of LL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying the plasmid pValac:BLG (LL-FnBPA+ BLG) was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Then we demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than the Caco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two different methods 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 without regard to the strains used (invasive or not); iii) the use of LL-FnBPA+ increased the number of mice producing BLG, but not the level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNA delivery vector in vivo.
Fil: Pontes, Daniela Santos. Institut National de la Recherche Agronomique; Francia
Fil: Innocentin, Silvia. Institut National de la Recherche Agronomique; Francia
Fil: del Carmen, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Franco Almeida, Juliana. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Leblanc, Jean Guy Joseph. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: de Moreno, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; Argentina
Fil: Blugeon, Sébastien. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Cherbuy, Claire. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Lefèvre, François. Institut National de la Recherche Agronomique; Francia
Fil: Azevedo, Vasco. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; Brasil
Fil: Miyoshi, Anderson. Universidade Federal Do Minas Gerais. Instituto de Cs.biologicas; Brasil
Fil: Langella, Philippe. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
Fil: Chatel, Jean-Marc. Institut National de la Recherche Agronomique; Francia. Centre de Cooperation Internationale En Recherche Agronomique Pour Le Developperment; Francia
description Lactococci are noninvasive lactic acid bacteria frequently used as protein delivery vectors and, more recently, as DNA delivery vehicles. We previously showed that Lactococcus lactis (LL) expressing the Fibronectin-Binding Protein A of Staphylococcus aureus (LL-FnBPA+) showed higher internalization rates in vitro in Caco-2 cells than the native (wt) lactococci and 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 of LL-FnBPA+ as an in vivo DNA vaccine delivery vehicle. We first showed that the invasive strain LL-FnBPA+ carrying the plasmid pValac:BLG (LL-FnBPA+ BLG) was more invasive than LL-BLG and showed the same invasivity as LL-FnBPA+. Then we demonstrated that the Caco-2 cells, co-incubated with LL-FnBPA+ BLG produced up to 30 times more BLG than the Caco-2 cells co-incubated with the non invasive LL-BLG. Using two different gene reporters, BLG and GFP, and two different methods 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 without regard to the strains used (invasive or not); iii) the use of LL-FnBPA+ increased the number of mice producing BLG, but not the level of BLG produced. We thus confirmed the good potential of invasive recombinant lactic acid bacteria as DNA delivery vector in vivo.
publishDate 2012
dc.date.none.fl_str_mv 2012-09-27
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/25289
Pontes, Daniela Santos; Innocentin, Silvia; del Carmen, Silvina Andrea; Franco Almeida, Juliana; Leblanc, Jean Guy Joseph; et al.; Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo; Public Library of Science; Plos One; 7; 9; 27-9-2012; 1-6; e44892-e44892
1932-6203
1932-6203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/25289
identifier_str_mv Pontes, Daniela Santos; Innocentin, Silvia; del Carmen, Silvina Andrea; Franco Almeida, Juliana; Leblanc, Jean Guy Joseph; et al.; Production of fibronectin binding protein A at the surface of lactococcus lactis increases plasmid transfer In vitro and in vivo; Public Library of Science; Plos One; 7; 9; 27-9-2012; 1-6; e44892-e44892
1932-6203
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0044892
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0044892
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
Atribución-NoComercial-CompartirIgual 2.5 Argentina (CC BY-NC-SA 2.5 AR)
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv Atribución-NoComercial-CompartirIgual 2.5 Argentina (CC BY-NC-SA 2.5 AR)
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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