Cargando…

Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors

Clostridium difficile induces antibiotic-associated diarrhea due to the release of toxin A (TcdA) and toxin B (TcdB), the latter being its main virulence factor. The epidemic strain NAP1/027 has an increased virulence attributed to different factors. We compared cellular intoxication by TcdB(NAP1) w...

Descripción completa

Detalles Bibliográficos
Autores principales: López-Ureña, Diana, Orozco-Aguilar, Josué, Chaves-Madrigal, Yendry, Ramírez-Mata, Andrea, Villalobos-Jimenez, Amanda, Ost, Stefan, Quesada-Gómez, Carlos, Rodríguez, César, Papatheodorou, Panagiotis, Chaves-Olarte, Esteban
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6628394/
https://www.ncbi.nlm.nih.gov/pubmed/31212980
http://dx.doi.org/10.3390/toxins11060348
_version_ 1783434949709266944
author López-Ureña, Diana
Orozco-Aguilar, Josué
Chaves-Madrigal, Yendry
Ramírez-Mata, Andrea
Villalobos-Jimenez, Amanda
Ost, Stefan
Quesada-Gómez, Carlos
Rodríguez, César
Papatheodorou, Panagiotis
Chaves-Olarte, Esteban
author_facet López-Ureña, Diana
Orozco-Aguilar, Josué
Chaves-Madrigal, Yendry
Ramírez-Mata, Andrea
Villalobos-Jimenez, Amanda
Ost, Stefan
Quesada-Gómez, Carlos
Rodríguez, César
Papatheodorou, Panagiotis
Chaves-Olarte, Esteban
author_sort López-Ureña, Diana
collection PubMed
description Clostridium difficile induces antibiotic-associated diarrhea due to the release of toxin A (TcdA) and toxin B (TcdB), the latter being its main virulence factor. The epidemic strain NAP1/027 has an increased virulence attributed to different factors. We compared cellular intoxication by TcdB(NAP1) with that by the reference strain VPI 10463 (TcdB(VPI)). In a mouse ligated intestinal loop model, TcdB(NAP1) induced higher neutrophil recruitment, cytokine release, and epithelial damage than TcdB(VPI). Both toxins modified the same panel of small GTPases and exhibited similar in vitro autoprocessing kinetics. On the basis of sequence variations in the frizzled-binding domain (FBD), we reasoned that TcdB(VPI) and TcdB(NAP1) might have different receptor specificities. To test this possibility, we used a TcdB from a NAP1 variant strain (TcdB(NAP1v)) unable to glucosylate RhoA but with the same receptor-binding domains as TcdB(NAP1). Cells were preincubated with TcdB(NAP1v) to block cellular receptors, prior to intoxication with either TcdB(VPI) or TcdB(NAP1). Preincubation with TcdB(NAP1v) blocked RhoA glucosylation by TcdB(NAP1) but not by TcdB(VPI), indicating that the toxins use different host factors for cell entry. This crucial difference might explain the increased biological activity of TcdB(NAP1) in the intestine, representing a contributing factor for the increased virulence of the NAP1/027 strain.
format Online
Article
Text
id pubmed-6628394
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-66283942019-07-23 Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors López-Ureña, Diana Orozco-Aguilar, Josué Chaves-Madrigal, Yendry Ramírez-Mata, Andrea Villalobos-Jimenez, Amanda Ost, Stefan Quesada-Gómez, Carlos Rodríguez, César Papatheodorou, Panagiotis Chaves-Olarte, Esteban Toxins (Basel) Article Clostridium difficile induces antibiotic-associated diarrhea due to the release of toxin A (TcdA) and toxin B (TcdB), the latter being its main virulence factor. The epidemic strain NAP1/027 has an increased virulence attributed to different factors. We compared cellular intoxication by TcdB(NAP1) with that by the reference strain VPI 10463 (TcdB(VPI)). In a mouse ligated intestinal loop model, TcdB(NAP1) induced higher neutrophil recruitment, cytokine release, and epithelial damage than TcdB(VPI). Both toxins modified the same panel of small GTPases and exhibited similar in vitro autoprocessing kinetics. On the basis of sequence variations in the frizzled-binding domain (FBD), we reasoned that TcdB(VPI) and TcdB(NAP1) might have different receptor specificities. To test this possibility, we used a TcdB from a NAP1 variant strain (TcdB(NAP1v)) unable to glucosylate RhoA but with the same receptor-binding domains as TcdB(NAP1). Cells were preincubated with TcdB(NAP1v) to block cellular receptors, prior to intoxication with either TcdB(VPI) or TcdB(NAP1). Preincubation with TcdB(NAP1v) blocked RhoA glucosylation by TcdB(NAP1) but not by TcdB(VPI), indicating that the toxins use different host factors for cell entry. This crucial difference might explain the increased biological activity of TcdB(NAP1) in the intestine, representing a contributing factor for the increased virulence of the NAP1/027 strain. MDPI 2019-06-17 /pmc/articles/PMC6628394/ /pubmed/31212980 http://dx.doi.org/10.3390/toxins11060348 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
López-Ureña, Diana
Orozco-Aguilar, Josué
Chaves-Madrigal, Yendry
Ramírez-Mata, Andrea
Villalobos-Jimenez, Amanda
Ost, Stefan
Quesada-Gómez, Carlos
Rodríguez, César
Papatheodorou, Panagiotis
Chaves-Olarte, Esteban
Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title_full Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title_fullStr Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title_full_unstemmed Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title_short Toxin B Variants from Clostridium difficile Strains VPI 10463 and NAP1/027 Share Similar Substrate Profile and Cellular Intoxication Kinetics but Use Different Host Cell Entry Factors
title_sort toxin b variants from clostridium difficile strains vpi 10463 and nap1/027 share similar substrate profile and cellular intoxication kinetics but use different host cell entry factors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6628394/
https://www.ncbi.nlm.nih.gov/pubmed/31212980
http://dx.doi.org/10.3390/toxins11060348
work_keys_str_mv AT lopezurenadiana toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT orozcoaguilarjosue toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT chavesmadrigalyendry toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT ramirezmataandrea toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT villalobosjimenezamanda toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT oststefan toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT quesadagomezcarlos toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT rodriguezcesar toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT papatheodoroupanagiotis toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors
AT chavesolarteesteban toxinbvariantsfromclostridiumdifficilestrainsvpi10463andnap1027sharesimilarsubstrateprofileandcellularintoxicationkineticsbutusedifferenthostcellentryfactors