Authors: 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
Publication Date: 2015.
Language: English.
Abstract:
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.
Author affiliation: Unger, Mandy. Universitat Hamburg; Alemania
Author affiliation: Eichhoff, Anna Marei. Universitat Hamburg; Alemania
Author affiliation: Schumacher, Lucas. Universitat Hamburg; Alemania
Author affiliation: Strysio, Moritz. Universitat Hamburg; Alemania
Author affiliation: Menzel, Stephan. Universitat Hamburg; Alemania
Author affiliation: Schwan, Carsten. University of Freiburg; Alemania
Author affiliation: 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
Author affiliation: Zylberman, Vanesa. Fundación Instituto Leloir; Argentina
Author affiliation: Seman, Michel. University Henry Poincare; Francia
Author affiliation: Brandner, Johanna. Universitat Hamburg; Alemania
Author affiliation: Rohde, Holger. Universitat Hamburg; Alemania
Author affiliation: Zhu, Kai. State University Of New York; Estados Unidos
Author affiliation: Haag, Friedrich. Universitat Hamburg; Alemania
Author affiliation: Mittrucker, Hans Willi. Universitat Hamburg; Alemania
Author affiliation: 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
Author affiliation: Aktories, Klaus. Universitat Hamburg; Alemania
Author affiliation: Koch Nolte, Friedrich. Universitat Hamburg; Alemania
Repository: CONICET Digital (CONICET). Consejo Nacional de Investigaciones Científicas y Técnicas