Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2

Autores
Bentancor, Leticia Veronica; Mejias, Maria Pilar; Pinto, Alípio; Bilen, Marcos Fabian; Meiss, Roberto; Rodriguez Galan, Maria Cecilia; Baez, Natalia Soledad; Pedrotti, Luciano Pablo; Goldstein Raij, Jorge; Ghiringhelli, Pablo Daniel; Palermo, Marina Sandra
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Shiga toxins (Stxs) are the main agent responsible for the development of hemolytic uremic syndrome (HUS), the most severe and life-threatening systemic complication of infection with enterohemorrhagic Escherichia coli (EHEC) strains. We previously reported Stx2 expression by eukaryotic cells after they were transfected in vitro with the stx2 gene cloned into a prokaryotic plasmid (pStx2). The aim of this study was to evaluate whether mammalian cells were also able to express Stx2 in vivo after pStx2 injection. Mice were inoculated by hydrodynamic based transfection (HBT) with pStx2. We studied the survival, the percentage of polymorphonuclear leukocytes in plasma, plasma urea levels and histology of the kidney and the brain of mice. Mice displayed a lethal dose-response to pStx2. Stx2-mRNA was recovered from the liver and Stx2 cytotoxic activity was observed in plasma of mice injected with pStx2. Stx2 was detected by immunofluorescence in the brains of mice inoculated with pStx2, and markers of central nervous system (CNS) damage were observed, including increased expression of glial fibrillary acidic protein (GFAP) and fragmentation of NeuN in neurons. Moreover, anti-Stx2B immunized mice were protected against pStx2 inoculation. Our results show that Stx2 is expressed in vivo from the wild stx2 gene, reproducing pathogenic damage induced by purified Stx2 or secondary to EHEC-infection.
Fil: Bentancor, Leticia Veronica. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Mejias, Maria Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Pinto, Alípio. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas; Argentina
Fil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina
Fil: Meiss, Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Rodriguez Galan, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Baez, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Pedrotti, Luciano Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Goldstein Raij, Jorge. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Ghiringhelli, Pablo Daniel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina
Fil: Palermo, Marina Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
Materia
Stxe
Promoter
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:11336/1913
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/1913
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2Bentancor, Leticia VeronicaMejias, Maria PilarPinto, AlípioBilen, Marcos FabianMeiss, RobertoRodriguez Galan, Maria CeciliaBaez, Natalia SoledadPedrotti, Luciano PabloGoldstein Raij, JorgeGhiringhelli, Pablo DanielPalermo, Marina SandraStxePromoterhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Shiga toxins (Stxs) are the main agent responsible for the development of hemolytic uremic syndrome (HUS), the most severe and life-threatening systemic complication of infection with enterohemorrhagic Escherichia coli (EHEC) strains. We previously reported Stx2 expression by eukaryotic cells after they were transfected in vitro with the stx2 gene cloned into a prokaryotic plasmid (pStx2). The aim of this study was to evaluate whether mammalian cells were also able to express Stx2 in vivo after pStx2 injection. Mice were inoculated by hydrodynamic based transfection (HBT) with pStx2. We studied the survival, the percentage of polymorphonuclear leukocytes in plasma, plasma urea levels and histology of the kidney and the brain of mice. Mice displayed a lethal dose-response to pStx2. Stx2-mRNA was recovered from the liver and Stx2 cytotoxic activity was observed in plasma of mice injected with pStx2. Stx2 was detected by immunofluorescence in the brains of mice inoculated with pStx2, and markers of central nervous system (CNS) damage were observed, including increased expression of glial fibrillary acidic protein (GFAP) and fragmentation of NeuN in neurons. Moreover, anti-Stx2B immunized mice were protected against pStx2 inoculation. Our results show that Stx2 is expressed in vivo from the wild stx2 gene, reproducing pathogenic damage induced by purified Stx2 or secondary to EHEC-infection.Fil: Bentancor, Leticia Veronica. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; ArgentinaFil: Mejias, Maria Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; ArgentinaFil: Pinto, Alípio. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas; ArgentinaFil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; ArgentinaFil: Meiss, Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; ArgentinaFil: Rodriguez Galan, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Baez, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Pedrotti, Luciano Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Goldstein Raij, Jorge. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; ArgentinaFil: Ghiringhelli, Pablo Daniel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; ArgentinaFil: Palermo, Marina Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaAmerican Society for Microbiology2013-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/1913Bentancor, Leticia Veronica; Mejias, Maria Pilar; Pinto, Alípio; Bilen, Marcos Fabian; Meiss, Roberto; et al.; Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2; American Society for Microbiology; mBio; 4; 5; 1-10-2013; 1-132150-7511enginfo:eu-repo/semantics/reference/url/info:eu-repo/semantics/reference es info:eu-repo/semantics/reference/pmid/24085779info:eu-repo/semantics/altIdentifier/doi/10.1128/mBio.00501-13info:eu-repo/semantics/altIdentifier/url/http://mbio.asm.org/content/4/5/e00501-13info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:53:02Zoai:ri.conicet.gov.ar:11336/1913instacron: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 09:53:02.699CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
title Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
spellingShingle Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
Bentancor, Leticia Veronica
Stxe
Promoter
title_short Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
title_full Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
title_fullStr Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
title_full_unstemmed Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
title_sort Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2
dc.creator.none.fl_str_mv Bentancor, Leticia Veronica
Mejias, Maria Pilar
Pinto, Alípio
Bilen, Marcos Fabian
Meiss, Roberto
Rodriguez Galan, Maria Cecilia
Baez, Natalia Soledad
Pedrotti, Luciano Pablo
Goldstein Raij, Jorge
Ghiringhelli, Pablo Daniel
Palermo, Marina Sandra
author Bentancor, Leticia Veronica
author_facet Bentancor, Leticia Veronica
Mejias, Maria Pilar
Pinto, Alípio
Bilen, Marcos Fabian
Meiss, Roberto
Rodriguez Galan, Maria Cecilia
Baez, Natalia Soledad
Pedrotti, Luciano Pablo
Goldstein Raij, Jorge
Ghiringhelli, Pablo Daniel
Palermo, Marina Sandra
author_role author
author2 Mejias, Maria Pilar
Pinto, Alípio
Bilen, Marcos Fabian
Meiss, Roberto
Rodriguez Galan, Maria Cecilia
Baez, Natalia Soledad
Pedrotti, Luciano Pablo
Goldstein Raij, Jorge
Ghiringhelli, Pablo Daniel
Palermo, Marina Sandra
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Stxe
Promoter
topic Stxe
Promoter
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Shiga toxins (Stxs) are the main agent responsible for the development of hemolytic uremic syndrome (HUS), the most severe and life-threatening systemic complication of infection with enterohemorrhagic Escherichia coli (EHEC) strains. We previously reported Stx2 expression by eukaryotic cells after they were transfected in vitro with the stx2 gene cloned into a prokaryotic plasmid (pStx2). The aim of this study was to evaluate whether mammalian cells were also able to express Stx2 in vivo after pStx2 injection. Mice were inoculated by hydrodynamic based transfection (HBT) with pStx2. We studied the survival, the percentage of polymorphonuclear leukocytes in plasma, plasma urea levels and histology of the kidney and the brain of mice. Mice displayed a lethal dose-response to pStx2. Stx2-mRNA was recovered from the liver and Stx2 cytotoxic activity was observed in plasma of mice injected with pStx2. Stx2 was detected by immunofluorescence in the brains of mice inoculated with pStx2, and markers of central nervous system (CNS) damage were observed, including increased expression of glial fibrillary acidic protein (GFAP) and fragmentation of NeuN in neurons. Moreover, anti-Stx2B immunized mice were protected against pStx2 inoculation. Our results show that Stx2 is expressed in vivo from the wild stx2 gene, reproducing pathogenic damage induced by purified Stx2 or secondary to EHEC-infection.
Fil: Bentancor, Leticia Veronica. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Mejias, Maria Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Pinto, Alípio. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas; Argentina
Fil: Bilen, Marcos Fabian. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina
Fil: Meiss, Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental; Argentina
Fil: Rodriguez Galan, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Baez, Natalia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Pedrotti, Luciano Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Goldstein Raij, Jorge. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Ciencias Fisiológicas. Laboratorio de Fisiopatogenia; Argentina
Fil: Ghiringhelli, Pablo Daniel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular; Argentina
Fil: Palermo, Marina Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina
description Shiga toxins (Stxs) are the main agent responsible for the development of hemolytic uremic syndrome (HUS), the most severe and life-threatening systemic complication of infection with enterohemorrhagic Escherichia coli (EHEC) strains. We previously reported Stx2 expression by eukaryotic cells after they were transfected in vitro with the stx2 gene cloned into a prokaryotic plasmid (pStx2). The aim of this study was to evaluate whether mammalian cells were also able to express Stx2 in vivo after pStx2 injection. Mice were inoculated by hydrodynamic based transfection (HBT) with pStx2. We studied the survival, the percentage of polymorphonuclear leukocytes in plasma, plasma urea levels and histology of the kidney and the brain of mice. Mice displayed a lethal dose-response to pStx2. Stx2-mRNA was recovered from the liver and Stx2 cytotoxic activity was observed in plasma of mice injected with pStx2. Stx2 was detected by immunofluorescence in the brains of mice inoculated with pStx2, and markers of central nervous system (CNS) damage were observed, including increased expression of glial fibrillary acidic protein (GFAP) and fragmentation of NeuN in neurons. Moreover, anti-Stx2B immunized mice were protected against pStx2 inoculation. Our results show that Stx2 is expressed in vivo from the wild stx2 gene, reproducing pathogenic damage induced by purified Stx2 or secondary to EHEC-infection.
publishDate 2013
dc.date.none.fl_str_mv 2013-10-01
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/11336/1913
Bentancor, Leticia Veronica; Mejias, Maria Pilar; Pinto, Alípio; Bilen, Marcos Fabian; Meiss, Roberto; et al.; Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2; American Society for Microbiology; mBio; 4; 5; 1-10-2013; 1-13
2150-7511
url http://hdl.handle.net/11336/1913
identifier_str_mv Bentancor, Leticia Veronica; Mejias, Maria Pilar; Pinto, Alípio; Bilen, Marcos Fabian; Meiss, Roberto; et al.; Promoter sequence of Shiga Toxin II (Stx2) is recognized in vivo leading to the production of biologically active Stx2; American Society for Microbiology; mBio; 4; 5; 1-10-2013; 1-13
2150-7511
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/reference/url/info:eu-repo/semantics/reference es info:eu-repo/semantics/reference/pmid/24085779
info:eu-repo/semantics/altIdentifier/doi/10.1128/mBio.00501-13
info:eu-repo/semantics/altIdentifier/url/http://mbio.asm.org/content/4/5/e00501-13
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.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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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score 13.070432

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