Comparison of the Caco-2, HT-29 and the mucus-secreting HT29-MTX intestinal cell models to investigate Salmonella adhesion and invasion

Gagnon, Mélanie; Berner, Annina Zihler; Chervet, Noémie; Chassard, Christophe; Lacroix, Christophe

doi:10.1016/j.mimet.2013.06.027

Cited by 213 publications

(169 citation statements)

References 40 publications

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“…Thus, hydrophobicity may be helpful for cell adhesion, but it is not a prerequisite for a strong adherence capacity (Schillinger et al, 2005) and thus not the only factor that determines bacterial adherence. Distinct binding rates of bacteria have also been previously documented in the case of different cell types (Gagnon et al, 2013).…”

Section: Hydrophobicity and Adherence Abilitiesmentioning

confidence: 74%

In vitro evaluation of candidate Bacillus spp. for animal feed

Mingmongkolchai

Panbangred

2017

J. Gen. Appl. Microbiol.

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The aim of this study was to select aerobic sporeformers for animal feed based on their in vitro probiotic potential, including their enzyme-producing ability and safety assessment. Seven isolates out of 187 spore-forming bacteria were selected for their ability to produce cellulase (89.21-1668.32 U/ ml), xylanase (1399.68-4351.10 U/ml), and phytase (2.72-28.70 U/ml). Among seven isolates, five had activities towards a broad range of p-nitrophenyl esters with acyl chain lengths from C2 to C12. The probiotic properties of all selected isolates varied with respect to their acid and bile salt tolerance under simulated gastrointestinal tract (GIT) conditions, and their adherence ability to human intestinal cell lines (Caco-2 and HT-29). The safety assessment revealed that the isolate CM40 was not cytotoxic to Caco-2 and HT-29, did not exhibit hemolytic activity, carried no enterotoxin or emetic toxin genes, and was susceptible to ten antibiotics, including six key antibiotics (chloramphenicol, erythromycin, gentamicin, tetracycline, streptomycin, and kanamycin) as recommended by the European Food Safety Authority (EFSA). Co-incubation of isolate CM40 with enteric bacteria (Salmonella Typhi, Salmonella Enteritidis 1781, and Escherichia coli) demonstrated that CM40 significantly decreased the number of pathogens (about 30-48%) adhering to Caco-2 and HT-29 (P < 0.05). Analysis of gene encoding 16S rRNA, gyrase A (gyrA) and the cheA histidine kinase revealed that CM40 belongs to Bacillus subtilis. On the basis of probiotic properties and basic safety aspects, the B. subtilis strain CM40 was found to possess desirable in vitro probiotic properties, and may be a potential candidate for supplementation of animal feed.

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Section: Hydrophobicity and Adherence Abilitiesmentioning

confidence: 74%

In vitro evaluation of candidate Bacillus spp. for animal feed

Mingmongkolchai

Panbangred

2017

J. Gen. Appl. Microbiol.

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“…28 Interestingly, it has also recently been reported that Salmonella virulence is enhanced in the presence of mucus. 29 On the other hand, H. pylori did not interact with HT29 cells and while it did interact with the mucin secreting HT29MTX cells, infection was markedly enhanced for E12 cells. Thus, either H. pylori increases expression of factors that mediate infection upon finding itself in an environment of mucus or, alternatively, the mucus layer offers an enhanced number of receptors that enables effective infection.…”

Section: Use Of Novel Tools To Assess the Interactions Of Bacteria Wimentioning

confidence: 93%

Interaction of microbes with mucus and mucins

et al. 2013

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Due to the recent rapid expansion in our understanding of the composition of the gut microflora and the consequences of altering that composition the question of how bacteria colonise mucus layers and interact with components of mucus, such as mucin, is now receiving widespread attention. Using a combination of mucus secreting cells, and a novel mucin microarray platform containing purified native mucins from different sources we recently demonstrated that two gastrointestinal pathogens, Helicobacter pylori and Campylobacter jejuni, colonise mucus by different mechanisms. This result emphasizes the potential for even closely related bacteria to interact with mucus in divergent ways to establish successful infection. Expanding the use of the mucin arrays described in the study to other microorganisms, both pathogenic and commensal, should lead to the discovery of biologically important motifs in bacterial-host interactions and complement the use of novel in vitro cell models, such as mucus secreting cell lines. IntroductionMucosal surfaces serve as a portal of entry for the majority of infections that occur in humans and animals. All mucosal surfaces are covered in a layer of mucus. In the gastrointestinal tract this mucus is colonised by tens of trillions of commensal organisms, essential for the development and health of the host. Mucus in the murine colon and the stomach has been shown to consist of an inner closely adherent mucus layer and an outer less dense, loosely adherent layer. 1,2The small intestine has a single loosely adherent layer.2 In the colon the outer mucus layer is colonised by commensal bacteria while the inner mucus layer is devoid of microbiota.1 The protective function of the inner mucus layer can be disrupted in disease, allowing access of organisms to the underlying epithelium. For example, the mucus layer has been shown to be depleted in animal models of colitis, and also in humans with ulcerative colitis resulting in bacterial penetration of the inner layer. 3A complex, mutually dependent relationship is established between the gut microbiota and the human host from birth which persists throughout life. 4 Initial cross talk between the host and gut microbiota influences postnatal intestinal epithelial differentiation by altering gene expression and improves immune tolerance. 5-7The microbiota also provides other benefits to the host including priming of the immune system and protection of the mucosal surface by competing with invading pathogens for space, nutrients, and binding receptors. Recent advances in sequencing technology and metagenomics have revealed changes in the microbial population of the gut in both health and disease. Strikingly, these studies have shown that alterations in the composition of the gut microflora, sometimes referred to as dysbiosis, are associated with a number of chronic diseases including obesity, diabetes and inflammatory disease. 8-10 Interaction of microbes with mucus and mucins Recent developmentsJulie Naughton, 1,2 Gina Duggan, 1, 2 Billy Bourke, 1,...

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“…1a, b). Nevertheless, Gagnon et al (2013) showed that the higher relative ability to invade was shown by S. Enteritidis. However, those authors found that Salmonella invasion using the HT-29 cell model is also dependent on the serovar, which seems to support the apparent contradiction associated with our results.…”

Section: Adhesion and Invasion Ability Of Strains To Ht-29 Cellsmentioning

confidence: 99%

“…1a). Gagnon et al (2013) showed that Salmonella adhesion to HT-29 cell depends on the serovar, with the higher relative abilities to adhere exhibited by S. Enteritidis. Our results confirm this finding and suggest that S. Enteritidis has a higher ability to adhere to the intestinal epithelium than S. Typhimurium.…”

Section: Adhesion and Invasion Ability Of Strains To Ht-29 Cellsmentioning

confidence: 99%

Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures

Dutra

Silva

Cabrita

et al. 2016

Journal of Medical Microbiology

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Listeria monocytogenes, Salmonella enterica and verocytotoxigenic Escherichia coli (VTEC) are amongst the most important agents responsible for food outbreaks occurring worldwide. In this work, two Lactobacillus spp. strains (LABs), Lactobacillus plantarum (LB95) and Lactobacillus paraplantarum (LB13), previously isolated from spontaneously fermenting olive brines, and two reference probiotic strains, Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, were investigated for their ability to attenuate the virulence of the aforementioned pathogens using animal cell culture assays. In competitive exclusion assays, the relative percentages of adhesion and invasion of S. enterica subsp. enterica serovar Enteritidis were significantly reduced when the human HT-29 cell line was previously exposed to LB95. The relative percentage of invasion by Listeria monocytogenes was significantly reduced when HT-29 cells were previously exposed to LB95. In the cytotoxicity assays, the cell-free supernatant of the co-culture (CFSC) of VTEC with LB95 accounted for the lowest value obtained amongst the co-cultures of VTEC with LABs, and was significantly lower than the value obtained with the co-culture of VTEC with the two probiotic reference strains. The cytotoxicity of CFSC of VTEC with both LB95 and LB13 exhibited values not significantly different from the cell-free supernatant of the nonpathogenic E. coli B strain. Our results suggested that LB95 may be able to attenuate the virulence of Gram-positive and Gram-negative food-borne pathogens; together with other reported features of these strains, our data reveal their possible use in probiotic foods due to their interesting potential in preventing enteric infections in humans.

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Comparison of the Caco-2, HT-29 and the mucus-secreting HT29-MTX intestinal cell models to investigate Salmonella adhesion and invasion

Cited by 213 publications

References 40 publications

<i>In vitro</i> evaluation of candidate <i>Bacillus</i> spp. for animal feed

<i>In vitro</i> evaluation of candidate <i>Bacillus</i> spp. for animal feed

Interaction of microbes with mucus and mucins

Lactobacillus plantarum LB95 impairs the virulence potential of Gram-positive and Gram-negative food-borne pathogens in HT-29 and Vero cell cultures

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