Cargando…

Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model

C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxi...

Descripción completa

Detalles Bibliográficos
Autores principales: Tonon, Federica, Di Bella, Stefano, Grassi, Gabriele, Luzzati, Roberto, Ascenzi, Paolo, di Masi, Alessandra, Zennaro, Cristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760802/
https://www.ncbi.nlm.nih.gov/pubmed/33271969
http://dx.doi.org/10.3390/cells9122575
_version_ 1783627418963017728
author Tonon, Federica
Di Bella, Stefano
Grassi, Gabriele
Luzzati, Roberto
Ascenzi, Paolo
di Masi, Alessandra
Zennaro, Cristina
author_facet Tonon, Federica
Di Bella, Stefano
Grassi, Gabriele
Luzzati, Roberto
Ascenzi, Paolo
di Masi, Alessandra
Zennaro, Cristina
author_sort Tonon, Federica
collection PubMed
description C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxin A (TcdA) and B (TcdB) produced by C. difficile have been studied in vivo using the zebrafish embryo model. Noteworthy, protective properties of human serum albumin (HSA) towards toxins-induced extra-intestinal effects were also addressed. Zebrafish embryos were treated with TcdA, TcdB and/or HSA at 24 h post-fertilization. Embryos were analyzed for 48 h after treatment to check vital signs and morphological changes. Markers related to cardio-vascular damage and inflammation were evaluated by Real-Time quantitative PCR and/or western blotting. Both toxins induced cardiovascular damage in zebrafish embryos by different mechanisms: (i) direct toxicity (i.e., pericardial edema, cardiac chambers enlargement, endothelial alteration); (ii) increased hormonal production and release (i.e., atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP)), (iii) alteration of the vascular system through the increase of the vascular endothelial growth factor (VEGF-A) levels, as well as of its receptors, (iv) pro-inflammatory response through high cytokines production (i.e., CXCL8, IL1B, IL6 and TNFα) and (v) cell-mediated damage due to the increase in neutrophils number. In addition to cardiovascular damage, we observe skin alteration and inflammation. Finally, our data indicate a protective effect of HSA toward the toxins induced extra-intestinal effects. Together, our findings can serve as a starting point for humans’ studies to substantiate and understand the extra-intestinal effects observed in CDI patients.
format Online
Article
Text
id pubmed-7760802
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-77608022020-12-26 Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model Tonon, Federica Di Bella, Stefano Grassi, Gabriele Luzzati, Roberto Ascenzi, Paolo di Masi, Alessandra Zennaro, Cristina Cells Article C.difficile infection (CDI) is not a merely “gut-confined” disease as toxemia could drive the development of CDI-related extra-intestinal effects. These effects could explain the high CDI-associated mortality, not just justified by diarrhea and dehydration. Here, the extra-intestinal effects of toxin A (TcdA) and B (TcdB) produced by C. difficile have been studied in vivo using the zebrafish embryo model. Noteworthy, protective properties of human serum albumin (HSA) towards toxins-induced extra-intestinal effects were also addressed. Zebrafish embryos were treated with TcdA, TcdB and/or HSA at 24 h post-fertilization. Embryos were analyzed for 48 h after treatment to check vital signs and morphological changes. Markers related to cardio-vascular damage and inflammation were evaluated by Real-Time quantitative PCR and/or western blotting. Both toxins induced cardiovascular damage in zebrafish embryos by different mechanisms: (i) direct toxicity (i.e., pericardial edema, cardiac chambers enlargement, endothelial alteration); (ii) increased hormonal production and release (i.e., atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP)), (iii) alteration of the vascular system through the increase of the vascular endothelial growth factor (VEGF-A) levels, as well as of its receptors, (iv) pro-inflammatory response through high cytokines production (i.e., CXCL8, IL1B, IL6 and TNFα) and (v) cell-mediated damage due to the increase in neutrophils number. In addition to cardiovascular damage, we observe skin alteration and inflammation. Finally, our data indicate a protective effect of HSA toward the toxins induced extra-intestinal effects. Together, our findings can serve as a starting point for humans’ studies to substantiate and understand the extra-intestinal effects observed in CDI patients. MDPI 2020-12-01 /pmc/articles/PMC7760802/ /pubmed/33271969 http://dx.doi.org/10.3390/cells9122575 Text en © 2020 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
Tonon, Federica
Di Bella, Stefano
Grassi, Gabriele
Luzzati, Roberto
Ascenzi, Paolo
di Masi, Alessandra
Zennaro, Cristina
Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title_full Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title_fullStr Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title_full_unstemmed Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title_short Extra-Intestinal Effects of C. difficile Toxin A and B: An In Vivo Study Using the Zebrafish Embryo Model
title_sort extra-intestinal effects of c. difficile toxin a and b: an in vivo study using the zebrafish embryo model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760802/
https://www.ncbi.nlm.nih.gov/pubmed/33271969
http://dx.doi.org/10.3390/cells9122575
work_keys_str_mv AT tononfederica extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT dibellastefano extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT grassigabriele extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT luzzatiroberto extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT ascenzipaolo extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT dimasialessandra extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel
AT zennarocristina extraintestinaleffectsofcdifficiletoxinaandbaninvivostudyusingthezebrafishembryomodel