Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages

Autores
Degarbo, S. M.; Grilli, Diego Javier; Telechea, A.; Lopez, M. F.; Arenas, G. N.
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
In recent years, this research team has progressed in the study of intracellular transport of the virulent strain (2308) of Brucella abortus in macrophage cell lines (J-774 and Raw) through multiple microscopic approaches. We demonstrated the transit of B. abortus 2308 through compartments of the endocytic pathway, marking early endosomes and lysosomes with gold (20 and 60 nm, respectively). We have shown that B. abortus 2308 occupies two different types of compartments: phagolysosomes and modified phagosomes, significantly reducing their fusion to endosomes. On the other hand, macrophages transfected with GFP-Rabs were used to evaluate the location of Rab 5 and 11 proteins, involved in the vesicular transport of the cell. Confocal microscopy showed that B. abortus 2308 (stained with Rhodamine) recruits Rab11 to the phagosome membrane that contains them, in the form of discrete patches. In addition, B. abortus 2308 co-locates with vesicles that overexpress Rab 5 in a large proportion. The permanence of Rab 5 and 11 associated with phagosomes containing B. abortus 2308 suggests that the bacteria actively retain these Rabs to avoid maturation of the phagosome that contains them. Subsequently, the effect of kinase inhibitors (AKTi) on the multiplication and intracellular survival of B. abortus 2308 at different times post macrophage infection was studied by confocal microscopy. These results allowed confirming that B. abortus 2308 uses the AKT/AS160 pathway to activate Rabs involved in the transport of nutrients necessary for its replication and the generation of a safe intracellular site. The compound used as an AKT inhibitor could constitute a new pharmacological approach for the treatment of brucellosis.
Fil: Degarbo, S. M.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Grilli, Diego Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Telechea, A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Lopez, M. F.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Arenas, G. N.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo
San Luis
Argentina
Sociedad de Biología de Cuyo
Materia
BRUCELLA ABORTUS
TRANSPORTE INTRACELULAR
MACRÓFAGOS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/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:11336/196138
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/196138
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophagesDegarbo, S. M.Grilli, Diego JavierTelechea, A.Lopez, M. F.Arenas, G. N.BRUCELLA ABORTUSTRANSPORTE INTRACELULARMACRÓFAGOShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In recent years, this research team has progressed in the study of intracellular transport of the virulent strain (2308) of Brucella abortus in macrophage cell lines (J-774 and Raw) through multiple microscopic approaches. We demonstrated the transit of B. abortus 2308 through compartments of the endocytic pathway, marking early endosomes and lysosomes with gold (20 and 60 nm, respectively). We have shown that B. abortus 2308 occupies two different types of compartments: phagolysosomes and modified phagosomes, significantly reducing their fusion to endosomes. On the other hand, macrophages transfected with GFP-Rabs were used to evaluate the location of Rab 5 and 11 proteins, involved in the vesicular transport of the cell. Confocal microscopy showed that B. abortus 2308 (stained with Rhodamine) recruits Rab11 to the phagosome membrane that contains them, in the form of discrete patches. In addition, B. abortus 2308 co-locates with vesicles that overexpress Rab 5 in a large proportion. The permanence of Rab 5 and 11 associated with phagosomes containing B. abortus 2308 suggests that the bacteria actively retain these Rabs to avoid maturation of the phagosome that contains them. Subsequently, the effect of kinase inhibitors (AKTi) on the multiplication and intracellular survival of B. abortus 2308 at different times post macrophage infection was studied by confocal microscopy. These results allowed confirming that B. abortus 2308 uses the AKT/AS160 pathway to activate Rabs involved in the transport of nutrients necessary for its replication and the generation of a safe intracellular site. The compound used as an AKT inhibitor could constitute a new pharmacological approach for the treatment of brucellosis.Fil: Degarbo, S. M.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Grilli, Diego Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Telechea, A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Lopez, M. F.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaFil: Arenas, G. N.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; ArgentinaXXXVII Reunión Científica Anual de la Sociedad de Biología de CuyoSan LuisArgentinaSociedad de Biología de CuyoTech Science Press2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/196138Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages; XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2020; 59-590327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://sbcuyo.org.ar/xxxv-reunion-cientifica-anual/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:04:11Zoai:ri.conicet.gov.ar:11336/196138instacron: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:34982025-09-03 10:04:11.266CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
title Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
spellingShingle Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
Degarbo, S. M.
BRUCELLA ABORTUS
TRANSPORTE INTRACELULAR
MACRÓFAGOS
title_short Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
title_full Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
title_fullStr Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
title_full_unstemmed Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
title_sort Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages
dc.creator.none.fl_str_mv Degarbo, S. M.
Grilli, Diego Javier
Telechea, A.
Lopez, M. F.
Arenas, G. N.
author Degarbo, S. M.
author_facet Degarbo, S. M.
Grilli, Diego Javier
Telechea, A.
Lopez, M. F.
Arenas, G. N.
author_role author
author2 Grilli, Diego Javier
Telechea, A.
Lopez, M. F.
Arenas, G. N.
author2_role author
author
author
author
dc.subject.none.fl_str_mv BRUCELLA ABORTUS
TRANSPORTE INTRACELULAR
MACRÓFAGOS
topic BRUCELLA ABORTUS
TRANSPORTE INTRACELULAR
MACRÓFAGOS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In recent years, this research team has progressed in the study of intracellular transport of the virulent strain (2308) of Brucella abortus in macrophage cell lines (J-774 and Raw) through multiple microscopic approaches. We demonstrated the transit of B. abortus 2308 through compartments of the endocytic pathway, marking early endosomes and lysosomes with gold (20 and 60 nm, respectively). We have shown that B. abortus 2308 occupies two different types of compartments: phagolysosomes and modified phagosomes, significantly reducing their fusion to endosomes. On the other hand, macrophages transfected with GFP-Rabs were used to evaluate the location of Rab 5 and 11 proteins, involved in the vesicular transport of the cell. Confocal microscopy showed that B. abortus 2308 (stained with Rhodamine) recruits Rab11 to the phagosome membrane that contains them, in the form of discrete patches. In addition, B. abortus 2308 co-locates with vesicles that overexpress Rab 5 in a large proportion. The permanence of Rab 5 and 11 associated with phagosomes containing B. abortus 2308 suggests that the bacteria actively retain these Rabs to avoid maturation of the phagosome that contains them. Subsequently, the effect of kinase inhibitors (AKTi) on the multiplication and intracellular survival of B. abortus 2308 at different times post macrophage infection was studied by confocal microscopy. These results allowed confirming that B. abortus 2308 uses the AKT/AS160 pathway to activate Rabs involved in the transport of nutrients necessary for its replication and the generation of a safe intracellular site. The compound used as an AKT inhibitor could constitute a new pharmacological approach for the treatment of brucellosis.
Fil: Degarbo, S. M.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Grilli, Diego Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Telechea, A.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Lopez, M. F.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
Fil: Arenas, G. N.. Universidad Nacional de Cuyo. Facultad de Cs.médicas. Departamento de Patología. Area de Microbiología; Argentina
XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo
San Luis
Argentina
Sociedad de Biología de Cuyo
description In recent years, this research team has progressed in the study of intracellular transport of the virulent strain (2308) of Brucella abortus in macrophage cell lines (J-774 and Raw) through multiple microscopic approaches. We demonstrated the transit of B. abortus 2308 through compartments of the endocytic pathway, marking early endosomes and lysosomes with gold (20 and 60 nm, respectively). We have shown that B. abortus 2308 occupies two different types of compartments: phagolysosomes and modified phagosomes, significantly reducing their fusion to endosomes. On the other hand, macrophages transfected with GFP-Rabs were used to evaluate the location of Rab 5 and 11 proteins, involved in the vesicular transport of the cell. Confocal microscopy showed that B. abortus 2308 (stained with Rhodamine) recruits Rab11 to the phagosome membrane that contains them, in the form of discrete patches. In addition, B. abortus 2308 co-locates with vesicles that overexpress Rab 5 in a large proportion. The permanence of Rab 5 and 11 associated with phagosomes containing B. abortus 2308 suggests that the bacteria actively retain these Rabs to avoid maturation of the phagosome that contains them. Subsequently, the effect of kinase inhibitors (AKTi) on the multiplication and intracellular survival of B. abortus 2308 at different times post macrophage infection was studied by confocal microscopy. These results allowed confirming that B. abortus 2308 uses the AKT/AS160 pathway to activate Rabs involved in the transport of nutrients necessary for its replication and the generation of a safe intracellular site. The compound used as an AKT inhibitor could constitute a new pharmacological approach for the treatment of brucellosis.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/196138
Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages; XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2020; 59-59
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/196138
identifier_str_mv Advances in the study of intracellular transport of brucela abortus 2308 (virulent strain) in macrophages; XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; San Luis; Argentina; 2020; 59-59
0327-9545
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://sbcuyo.org.ar/xxxv-reunion-cientifica-anual/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
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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