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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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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 |
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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 |
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