A Novel Strategy to Study the Invasive Capability of Adherent-Invasive Escherichia coli by Using Human Primary Organoid-Derived Epithelial Monolayers
| Autor: | Queralt Bonet-Rossinyol, Isabella Dotti, Marta Martínez-Picola, Azucena Salas, Miriam Esteller, Margarita Martinez-Medina, Aida Mayorgas, Elena Ricart |
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| Přispěvatelé: | Agencia Estatal de Investigación |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
lcsh:Immunologic diseases. Allergy Immunology Biology Crohn Malaltia de Sistema immunològic Microbiology Pathogenesis 03 medical and health sciences 0302 clinical medicine Immune system Organoid Escherichia coli Immunology and Allergy Pathogen organoid-derived epithelial monolayers (ODM) bacterial infection intestinal epithelial cells (IECs) Intestinal epithelium inflammatory bowel disease (IBD) 3. Good health Crohn Malaltia de -- Patogènesi Crohn's disease Crohn's disease -- Pathogenesis 030104 developmental biology adherent-invasive E. coli (AIEC) Escheríchia coli Cell culture 030211 gastroenterology & hepatology lcsh:RC581-607 Immortalised cell line Intracellular |
| Zdroj: | Frontiers in Immunology, 2021, vol. 12, art.núm. 646906 Articles publicats (D-B) DUGiDocs – Universitat de Girona instname Frontiers in Immunology, Vol 12 (2021) |
| Popis: | 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 AM was supported by the Programa Estatal de Investigación, Desarrollo e Innovación del Ministerio de Economía, Industria y Competitividad, co-funded by the European Social Fund (ESF) under grant agreement number FPI BES-2016-076642. ID is funded by the Horizon 2020 Framework Programme (EU for Research and Innovation H2020), Grant 720905. QB-R was supported by the Programa d’Ajuts per a Investigadors en Formació de la Universitat de Girona – IFUdG 2019/21. AS is funded by the Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD) and Grant RTI2018- 096946-B-I00 from the Ministerio de Ciencia, Innovación y Universidades, Spain. MM-M is funded by the Spanish Ministry of Economy and Competitiveness through project SAF2017-82261-P, which is co-funded by the European Regional Development Fund |
| Databáze: | OpenAIRE |
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