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A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers

Over the last decades, Adherent-Invasive Escherichia coli (AIEC) has been linked to the pathogenesis of Crohn’s Disease. AIEC’s characteristics, as well as its interaction with the gut immune system and its role in intestinal epithelial barrier dysfunction, have been extensively studied. Nevertheles...

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Autores principales: Mayorgas, Aida, Dotti, Isabella, Martínez-Picola, Marta, Esteller, Miriam, Bonet-Rossinyol, Queralt, Ricart, Elena, Salas, Azucena, Martínez-Medina, Margarita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8039293/
https://www.ncbi.nlm.nih.gov/pubmed/33854511
http://dx.doi.org/10.3389/fimmu.2021.646906
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author Mayorgas, Aida
Dotti, Isabella
Martínez-Picola, Marta
Esteller, Miriam
Bonet-Rossinyol, Queralt
Ricart, Elena
Salas, Azucena
Martínez-Medina, Margarita
author_facet Mayorgas, Aida
Dotti, Isabella
Martínez-Picola, Marta
Esteller, Miriam
Bonet-Rossinyol, Queralt
Ricart, Elena
Salas, Azucena
Martínez-Medina, Margarita
author_sort Mayorgas, Aida
collection PubMed
description Over the last decades, Adherent-Invasive Escherichia coli (AIEC) has been linked to the pathogenesis of Crohn’s Disease. AIEC’s characteristics, as well as its interaction with the gut immune system and its role in intestinal epithelial barrier dysfunction, have been extensively studied. Nevertheless, the currently available techniques to investigate the cross-talk between this pathogen and intestinal epithelial cells (IECs) are based on the infection of immortalized cell lines. Despite their many advantages, cell lines cannot reproduce the conditions in tissues, nor do they reflect interindividual variability or gut location-specific traits. In that sense, the use of human primary cultures, either healthy or diseased, offers a system that can overcome all of these limitations. Here, we developed a new infection model by using freshly isolated human IECs. For the first time, we generated and infected monolayer cultures derived from human colonic organoids to study the mechanisms and effects of AIEC adherence and invasion on primary human epithelial cells. To establish the optimal conditions for AIEC invasion studies in human primary organoid-derived epithelial monolayers, we designed an infection-kinetics study to assess the infection dynamics at different time points, as well as with two multiplicities of infection (MOI). Overall, this method provides a model for the study of host response to AIEC infections, as well as for the understanding of the molecular mechanisms involved in adhesion, invasion and intracellular replication. Therefore, it represents a promising tool for elucidating the cross-talk between AIEC and the intestinal epithelium in healthy and diseased tissues.
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spelling pubmed-80392932021-04-13 A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers Mayorgas, Aida Dotti, Isabella Martínez-Picola, Marta Esteller, Miriam Bonet-Rossinyol, Queralt Ricart, Elena Salas, Azucena Martínez-Medina, Margarita Front Immunol Immunology Over the last decades, Adherent-Invasive Escherichia coli (AIEC) has been linked to the pathogenesis of Crohn’s Disease. AIEC’s characteristics, as well as its interaction with the gut immune system and its role in intestinal epithelial barrier dysfunction, have been extensively studied. Nevertheless, the currently available techniques to investigate the cross-talk between this pathogen and intestinal epithelial cells (IECs) are based on the infection of immortalized cell lines. Despite their many advantages, cell lines cannot reproduce the conditions in tissues, nor do they reflect interindividual variability or gut location-specific traits. In that sense, the use of human primary cultures, either healthy or diseased, offers a system that can overcome all of these limitations. Here, we developed a new infection model by using freshly isolated human IECs. For the first time, we generated and infected monolayer cultures derived from human colonic organoids to study the mechanisms and effects of AIEC adherence and invasion on primary human epithelial cells. To establish the optimal conditions for AIEC invasion studies in human primary organoid-derived epithelial monolayers, we designed an infection-kinetics study to assess the infection dynamics at different time points, as well as with two multiplicities of infection (MOI). Overall, this method provides a model for the study of host response to AIEC infections, as well as for the understanding of the molecular mechanisms involved in adhesion, invasion and intracellular replication. Therefore, it represents a promising tool for elucidating the cross-talk between AIEC and the intestinal epithelium in healthy and diseased tissues. Frontiers Media S.A. 2021-03-29 /pmc/articles/PMC8039293/ /pubmed/33854511 http://dx.doi.org/10.3389/fimmu.2021.646906 Text en Copyright © 2021 Mayorgas, Dotti, Martínez-Picola, Esteller, Bonet-Rossinyol, Ricart, Salas and Martínez-Medina https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Mayorgas, Aida
Dotti, Isabella
Martínez-Picola, Marta
Esteller, Miriam
Bonet-Rossinyol, Queralt
Ricart, Elena
Salas, Azucena
Martínez-Medina, Margarita
A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title_full A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title_fullStr A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title_full_unstemmed A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title_short A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
title_sort novel strategy to study the invasive capability of adherent-invasive escherichia coli by using human primary organoid-derived epithelial monolayers
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8039293/
https://www.ncbi.nlm.nih.gov/pubmed/33854511
http://dx.doi.org/10.3389/fimmu.2021.646906
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