Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT

Autores
Unger, Mandy; Eichhoff, Anna Marei; Schumacher, Lucas; Strysio, Moritz; Menzel, Stephan; Schwan, Carsten; Alzogaray, Vanina Andrea; Zylberman, Vanesa; Seman, Michel; Brandner, Johanna; Rohde, Holger; Zhu, Kai; Haag, Friedrich; Mittrucker, Hans Willi; Goldbaum, Fernando Alberto; Aktories, Klaus; Koch Nolte, Friedrich
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The spore-forming gut bacterium Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitalized patients. The major virulence factors are two large glucosylating cytotoxins. Hypervirulent strains (e.g. ribotype 027) with higher morbidity and mortality additionally produce the binary CDT toxin (Clostridium difficile transferase) that ADP-ribosylates actin and induces microtubule-based cell protrusions. Nanobodies are robust single domain antibodies derived from camelid heavy chain antibodies. Here we report the generation of functional nanobodies against the enzymatic CDTa and the heptameric receptor binding subunit CDTb. The nanobodies were obtained from a variable-domain repertoire library isolated from llamas immunized with recombinant CDTa or CDTb. Five CDTa-specific nanobodies blocked CDTa-mediated ADP-ribosylation of actin. Three CDTa-specific and two CDTb-specific nanobodies neutralized the cytotoxicity of CDTa+b. These nanobodies hold promise as new tools for research, diagnosis and therapy of C. difficile associated disease.
Fil: Unger, Mandy. Universitat Hamburg; Alemania
Fil: Eichhoff, Anna Marei. Universitat Hamburg; Alemania
Fil: Schumacher, Lucas. Universitat Hamburg; Alemania
Fil: Strysio, Moritz. Universitat Hamburg; Alemania
Fil: Menzel, Stephan. Universitat Hamburg; Alemania
Fil: Schwan, Carsten. University of Freiburg; Alemania
Fil: Alzogaray, Vanina Andrea. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina
Fil: Zylberman, Vanesa. Fundación Instituto Leloir; Argentina
Fil: Seman, Michel. University Henry Poincare; Francia
Fil: Brandner, Johanna. Universitat Hamburg; Alemania
Fil: Rohde, Holger. Universitat Hamburg; Alemania
Fil: Zhu, Kai. State University Of New York; Estados Unidos
Fil: Haag, Friedrich. Universitat Hamburg; Alemania
Fil: Mittrucker, Hans Willi. Universitat Hamburg; Alemania
Fil: Goldbaum, Fernando Alberto. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina
Fil: Aktories, Klaus. Universitat Hamburg; Alemania
Fil: Koch Nolte, Friedrich. Universitat Hamburg; Alemania
Materia
Nanobodies
Cdt Toxin
Clostridium Difficile
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:11336/9651
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/9651
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDTUnger, MandyEichhoff, Anna MareiSchumacher, LucasStrysio, MoritzMenzel, StephanSchwan, CarstenAlzogaray, Vanina AndreaZylberman, VanesaSeman, MichelBrandner, JohannaRohde, HolgerZhu, KaiHaag, FriedrichMittrucker, Hans WilliGoldbaum, Fernando AlbertoAktories, KlausKoch Nolte, FriedrichNanobodiesCdt ToxinClostridium Difficilehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The spore-forming gut bacterium Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitalized patients. The major virulence factors are two large glucosylating cytotoxins. Hypervirulent strains (e.g. ribotype 027) with higher morbidity and mortality additionally produce the binary CDT toxin (Clostridium difficile transferase) that ADP-ribosylates actin and induces microtubule-based cell protrusions. Nanobodies are robust single domain antibodies derived from camelid heavy chain antibodies. Here we report the generation of functional nanobodies against the enzymatic CDTa and the heptameric receptor binding subunit CDTb. The nanobodies were obtained from a variable-domain repertoire library isolated from llamas immunized with recombinant CDTa or CDTb. Five CDTa-specific nanobodies blocked CDTa-mediated ADP-ribosylation of actin. Three CDTa-specific and two CDTb-specific nanobodies neutralized the cytotoxicity of CDTa+b. These nanobodies hold promise as new tools for research, diagnosis and therapy of C. difficile associated disease.Fil: Unger, Mandy. Universitat Hamburg; AlemaniaFil: Eichhoff, Anna Marei. Universitat Hamburg; AlemaniaFil: Schumacher, Lucas. Universitat Hamburg; AlemaniaFil: Strysio, Moritz. Universitat Hamburg; AlemaniaFil: Menzel, Stephan. Universitat Hamburg; AlemaniaFil: Schwan, Carsten. University of Freiburg; AlemaniaFil: Alzogaray, Vanina Andrea. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; ArgentinaFil: Zylberman, Vanesa. Fundación Instituto Leloir; ArgentinaFil: Seman, Michel. University Henry Poincare; FranciaFil: Brandner, Johanna. Universitat Hamburg; AlemaniaFil: Rohde, Holger. Universitat Hamburg; AlemaniaFil: Zhu, Kai. State University Of New York; Estados UnidosFil: Haag, Friedrich. Universitat Hamburg; AlemaniaFil: Mittrucker, Hans Willi. Universitat Hamburg; AlemaniaFil: Goldbaum, Fernando Alberto. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; ArgentinaFil: Aktories, Klaus. Universitat Hamburg; AlemaniaFil: Koch Nolte, Friedrich. Universitat Hamburg; AlemaniaNature Publishing Group2015-01info: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/11336/9651Unger, Mandy; Eichhoff, Anna Marei; Schumacher, Lucas; Strysio, Moritz; Menzel, Stephan; et al.; Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT; Nature Publishing Group; Scientific Reports; 5; 7850; 1-2015; 1-102045-2322enginfo:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/srep07850info:eu-repo/semantics/altIdentifier/doi/10.1038/srep07850info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:57:47Zoai:ri.conicet.gov.ar:11336/9651instacron: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 09:57:47.8CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
title Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
spellingShingle Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
Unger, Mandy
Nanobodies
Cdt Toxin
Clostridium Difficile
title_short Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
title_full Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
title_fullStr Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
title_full_unstemmed Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
title_sort Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT
dc.creator.none.fl_str_mv Unger, Mandy
Eichhoff, Anna Marei
Schumacher, Lucas
Strysio, Moritz
Menzel, Stephan
Schwan, Carsten
Alzogaray, Vanina Andrea
Zylberman, Vanesa
Seman, Michel
Brandner, Johanna
Rohde, Holger
Zhu, Kai
Haag, Friedrich
Mittrucker, Hans Willi
Goldbaum, Fernando Alberto
Aktories, Klaus
Koch Nolte, Friedrich
author Unger, Mandy
author_facet Unger, Mandy
Eichhoff, Anna Marei
Schumacher, Lucas
Strysio, Moritz
Menzel, Stephan
Schwan, Carsten
Alzogaray, Vanina Andrea
Zylberman, Vanesa
Seman, Michel
Brandner, Johanna
Rohde, Holger
Zhu, Kai
Haag, Friedrich
Mittrucker, Hans Willi
Goldbaum, Fernando Alberto
Aktories, Klaus
Koch Nolte, Friedrich
author_role author
author2 Eichhoff, Anna Marei
Schumacher, Lucas
Strysio, Moritz
Menzel, Stephan
Schwan, Carsten
Alzogaray, Vanina Andrea
Zylberman, Vanesa
Seman, Michel
Brandner, Johanna
Rohde, Holger
Zhu, Kai
Haag, Friedrich
Mittrucker, Hans Willi
Goldbaum, Fernando Alberto
Aktories, Klaus
Koch Nolte, Friedrich
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Nanobodies
Cdt Toxin
Clostridium Difficile
topic Nanobodies
Cdt Toxin
Clostridium Difficile
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The spore-forming gut bacterium Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitalized patients. The major virulence factors are two large glucosylating cytotoxins. Hypervirulent strains (e.g. ribotype 027) with higher morbidity and mortality additionally produce the binary CDT toxin (Clostridium difficile transferase) that ADP-ribosylates actin and induces microtubule-based cell protrusions. Nanobodies are robust single domain antibodies derived from camelid heavy chain antibodies. Here we report the generation of functional nanobodies against the enzymatic CDTa and the heptameric receptor binding subunit CDTb. The nanobodies were obtained from a variable-domain repertoire library isolated from llamas immunized with recombinant CDTa or CDTb. Five CDTa-specific nanobodies blocked CDTa-mediated ADP-ribosylation of actin. Three CDTa-specific and two CDTb-specific nanobodies neutralized the cytotoxicity of CDTa+b. These nanobodies hold promise as new tools for research, diagnosis and therapy of C. difficile associated disease.
Fil: Unger, Mandy. Universitat Hamburg; Alemania
Fil: Eichhoff, Anna Marei. Universitat Hamburg; Alemania
Fil: Schumacher, Lucas. Universitat Hamburg; Alemania
Fil: Strysio, Moritz. Universitat Hamburg; Alemania
Fil: Menzel, Stephan. Universitat Hamburg; Alemania
Fil: Schwan, Carsten. University of Freiburg; Alemania
Fil: Alzogaray, Vanina Andrea. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina
Fil: Zylberman, Vanesa. Fundación Instituto Leloir; Argentina
Fil: Seman, Michel. University Henry Poincare; Francia
Fil: Brandner, Johanna. Universitat Hamburg; Alemania
Fil: Rohde, Holger. Universitat Hamburg; Alemania
Fil: Zhu, Kai. State University Of New York; Estados Unidos
Fil: Haag, Friedrich. Universitat Hamburg; Alemania
Fil: Mittrucker, Hans Willi. Universitat Hamburg; Alemania
Fil: Goldbaum, Fernando Alberto. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina
Fil: Aktories, Klaus. Universitat Hamburg; Alemania
Fil: Koch Nolte, Friedrich. Universitat Hamburg; Alemania
description The spore-forming gut bacterium Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitalized patients. The major virulence factors are two large glucosylating cytotoxins. Hypervirulent strains (e.g. ribotype 027) with higher morbidity and mortality additionally produce the binary CDT toxin (Clostridium difficile transferase) that ADP-ribosylates actin and induces microtubule-based cell protrusions. Nanobodies are robust single domain antibodies derived from camelid heavy chain antibodies. Here we report the generation of functional nanobodies against the enzymatic CDTa and the heptameric receptor binding subunit CDTb. The nanobodies were obtained from a variable-domain repertoire library isolated from llamas immunized with recombinant CDTa or CDTb. Five CDTa-specific nanobodies blocked CDTa-mediated ADP-ribosylation of actin. Three CDTa-specific and two CDTb-specific nanobodies neutralized the cytotoxicity of CDTa+b. These nanobodies hold promise as new tools for research, diagnosis and therapy of C. difficile associated disease.
publishDate 2015
dc.date.none.fl_str_mv 2015-01
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/11336/9651
Unger, Mandy; Eichhoff, Anna Marei; Schumacher, Lucas; Strysio, Moritz; Menzel, Stephan; et al.; Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT; Nature Publishing Group; Scientific Reports; 5; 7850; 1-2015; 1-10
2045-2322
url http://hdl.handle.net/11336/9651
identifier_str_mv Unger, Mandy; Eichhoff, Anna Marei; Schumacher, Lucas; Strysio, Moritz; Menzel, Stephan; et al.; Selection of nanobodies that block the enzymatic and cytotoxic activities of the binary Clostridium difficile toxin CDT; Nature Publishing Group; Scientific Reports; 5; 7850; 1-2015; 1-10
2045-2322
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/srep07850
info:eu-repo/semantics/altIdentifier/doi/10.1038/srep07850
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
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
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 13.13397

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