Experimental models to study intestinal microbes–mucus interactions in health and disease

Etienne‐Mesmin, Lucie; Chassaing, Benoît; Desvaux, Mickaël; Paepe, Kim De; Gresse, Raphaële; Sauvaitre, Thomas; Forano, Evelyne; Wiele, Tom Van de; Schüller, Stephanie; Juge, Nathalie; Blanquet‐Diot, Stéphanie

doi:10.1093/femsre/fuz013

Cited by 125 publications

(105 citation statements)

References 364 publications

(407 reference statements)

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“…In the presence of a eukaryotic host, the performance and behavior of the phage will likely vary significantly. Here we describe the interaction of phage A2 and its host bacterium E. faecalis with respect to a mucus producing intestinal epithelial layer, better mimicking the conditions found in the mucosal layer of the human gut [ 49 ]. In this model, we could judge the potential therapeutic value of the phage as well as understand the translocation, adhesion, and invasion of both phage and target bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…For the long-term experiments cells were seeded at a density of 15,000 cells/cm 2 for the long-term experiments and incubated in growth media for 20 days. Under these conditions, mucus-producing Ht29-MTX cells are able to produce a continuous mucus layer [ 49 ]. Once the cultures were ready to treat, the medium was removed and cells were washed with Dulbecco’s phosphate buffer saline (DPBS) before adding fresh antibiotic-free medium containing 10 6 CFU/mL E. faecalis .…”

Section: Methodsmentioning

confidence: 99%

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Characterizing Phage-Host Interactions in a Simplified Human Intestinal Barrier Model

et al. 2020

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An intestinal epithelium model able to produce mucus was developed to provide an environment suitable for testing the therapeutic activity of gut bacteriophages. We show that Enterococcus faecalis adheres more effectively in the presence of mucus, can invade the intestinal epithelia and is able to translocate after damaging tight junctions. Furthermore, Enterococcus phage vB_EfaM_A2 (a member of Herelleviridae that possesses virion associated immunoglobin domains) was found to translocate through the epithelium in the presence and absence of its host bacteria. Phage A2 protected eukaryotic cells by reducing mortality and maintaining the structure of the cell layer structure. We suggest the mammalian cell model utilized within this study as an adaptable in vitro model that can be employed to enable a better understanding of phage–bacteria interactions and the protective impact of phage therapy relating to the intestinal epithelium.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Characterizing Phage-Host Interactions in a Simplified Human Intestinal Barrier Model

et al. 2020

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“…Salmonella Typhimurium actively degrades the glycocalyx during infection, a strategy suspected to promote long lasting attachment 41 . Interestingly, bacterial adhesins in some instances target surface glycans directly rather than proteins, transforming the glycocalyx shield into the target itself 42 . We have shown flagella also favor bacterial release, potentially providing an explanation for flagellum shedding it upon attachment 43 .…”

Section: Discussionmentioning

confidence: 99%

The membrane microenvironment regulates the sequential attachment of bacteria to host cells

Pierrat

Wong

Al-Mayyah

et al. 2020

Preprint

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Pathogen attachment to host tissue is critical in the progress of many infections. Bacteria use adhesion in vivo to promote colonization and regulate the deployment of contact-dependent virulence traits. To specifically target host cells, they decorate themselves with adhesins, proteins that bind to mammalian cell surface receptors. One common assumption is that adhesin-receptor interactions entirely govern bacterial attachment. However, how adhesins engage with their receptors in an in vivo-like context remains unclear, in particular under the influence of a heterogeneous mechanical microenvironment. We here investigate the biophysical processes governing bacterial adhesion to host cells using a tunable adhesin-receptor system. By dynamically visualizing attachment, we found that bacterial adhesion to host cell surface, unlike adhesion to inert surfaces, involves two consecutive steps. Bacteria initially attach to their host without engaging adhesins. This step lasts about one minute during which bacteria can easily detach. We found that at this stage, the glycocalyx, a layer of glycosylated proteins and lipids, shields the host cell by keeping adhesin away from their receptor ligand. In a second step, adhesins engage with their target receptors to strengthen attachment for minutes to hours. The active properties of the membrane, endowed by the actin cytoskeleton, strengthen specific adhesion. Altogether, our results demonstrate that adhesin-ligand binding is not the sole regulator of bacterial adhesion. In fact, the host cell’s mechanical microenvironment relatively strongly mediated host-bacteria physical interactions, thereby playing an essential role in the onset of infection.

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“…However, when these cells are grown in media where galactose replaces glucose, the cells can be modulated to produce mucin and the HT29-C18N2 clone has therefore been used as a model system for goblet cell differentiation [172]. Other mucin-producing clones have emerged from the parental cell line HT29 after subculture with sodium butyrate or 5-fluorouracil [173,174]. In contrast to other HT29 clones, the HT29-MTX cell line are induced to produce relatively high levels of mucin by stepwise adaptation to increasing concentrations of methotrexate (MTX) [173,175].…”

Section: Ht29 Cell Linementioning

confidence: 99%

The Intestinal Barrier and Current Techniques for the Assessment of Gut Permeability

Schoultz

Keita

2020

Cells

243

222

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The intestinal barrier is essential in human health and constitutes the interface between the outside and the internal milieu of the body. A functional intestinal barrier allows absorption of nutrients and fluids but simultaneously prevents harmful substances like toxins and bacteria from crossing the intestinal epithelium and reaching the body. An altered intestinal permeability, a sign of a perturbed barrier function, has during the last decade been associated with several chronic conditions, including diseases originating in the gastrointestinal tract but also diseases such as Alzheimer and Parkinson disease. This has led to an intensified interest from researchers with diverse backgrounds to perform functional studies of the intestinal barrier in different conditions. Intestinal permeability is defined as the passage of a solute through a simple membrane and can be measured by recording the passage of permeability markers over the epithelium via the paracellular or the transcellular route. The methodological tools to investigate the gut barrier function are rapidly expanding and new methodological approaches are being developed. Here we outline and discuss, in vivo, in vitro and ex vivo techniques and how these methods can be utilized for thorough investigation of the intestinal barrier.

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Experimental models to study intestinal microbes–mucus interactions in health and disease

Cited by 125 publications

References 364 publications

Characterizing Phage-Host Interactions in a Simplified Human Intestinal Barrier Model

Characterizing Phage-Host Interactions in a Simplified Human Intestinal Barrier Model

The membrane microenvironment regulates the sequential attachment of bacteria to host cells

The Intestinal Barrier and Current Techniques for the Assessment of Gut Permeability

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