scholarly journals Mannan rich fraction from yeast modulates inflammatory responses in intestinal cells (HT-29) exposed toEscherichia coli

2019 ◽  
Vol 7 ◽  
Author(s):  
Niall Browne ◽  
Aimee Traynor ◽  
Karina A. Horgan
Keyword(s):  
Inflammatory Responses ◽  
Economic Cost ◽  
Intestinal Cell ◽  
Cell Protein ◽  
Intestinal Cells ◽  
E Coli ◽  
Intestinal Cell Line ◽  
Proinflammatory Responses ◽  
Ht 29

AbstractMannan from yeast has been demonstrated to limit infection in animals susceptible to gastrointestinal infection, including pigs, poultry and cows, by blocking the mechanism by which gram-negative bacteria adhere to and invade the intestines. EnterotoxigenicEscherichia coli(ETEC) cause post weaning diarrhoea (PWD) which results in poor weight gain and potential death at great economic cost to the farmer. A mannan rich fraction (MRF) was assessedin vitrofor its impact on ETEC infection of HT-29 intestinal cell line. Gene expression markers for inflammation (TNFαandIL-1β) and TLR4 (TICAM-1andLY96) associated recognition of bacteria were significantly elevated following exposure toE. colialone, but not in combination with MRF compared to the control. HT-29 cells exposed to MRF alone demonstrated significantly reduced expression of immune signalling genesIRAK1,IRF7andJUNwhen compared to the control. HT-29 cell protein abundance for TNFα and TLR4 associated proteins were significantly increased in response toE. coliexposure alone while no significant change was observed for MRF treatment withE. coliinfection.E. coliadhesion to HT-29 cells was significantly decreased with addition of MRF compared toE. coliinfection alone. The action of MRF demonstrated its potential capacity to limit infection on anin vitrolevel through blocking bacterial interaction with the intestines that leads to infection as marked by a reduction in proinflammatory responses. MRF on its own demonstrated potential anti-inflammatory effects on intestinal cells with the reduction of proinflammatory responses observed.

scholarly journals Preliminary Characterization of the Transcriptional Response of the Porcine Intestinal Cell Line IPEC-J2 to EnterotoxigenicEscherichia coli,Escherichia coli, andE. coliLipopolysaccharide

2010 ◽  
Vol 2010 ◽  
pp. 1-11 ◽  
Author(s):  
Marisa M. Geens ◽  
Theo A. Niewold
Keyword(s):  
Escherichia Coli ◽  
Cell Line ◽  
Transcriptional Response ◽  
Intestinal Cell ◽  
Adhesion Assay ◽  
Transcriptional Level ◽  
Suitable Model ◽  
E Coli ◽  
Intestinal Cell Line

IPEC-J2, a promisingin vitromodel system, is not well characterized especially on the transcriptional level, in contrast to human counterparts. The aim of this study was to characterize the gene expression in IPEC-J2 cells when coincubated with enterotoxigenicEscherichia coli(ETEC), nonpathogenicE. coli, andE. coliendotoxin. Apical infection of polarized IPEC-J2 monolayers caused a time-dependent decrease in transepithelial electrical resistance (TEER). Microarray analysis showed up-regulation of interleukins when IPEC-J2 were cocultured withE. colistrains this has so far never been measured in this cell line. Highest IL8 expression was found with the ETEC strain possessing the F4 fimbrium, suggesting IPEC-J2 cells to be F4 receptor positive, confirmed in a brush border membrane adhesion assay. It is concluded that the innate immune responses to pathogens and LPS makes the IPEC-J2 cell line a suitable model for research on intestinal host pathogen interaction.

scholarly journals Acute and Sub-Chronic Effects of Microplastics (3 and 10 µm) on the Human Intestinal Cells HT-29

2021 ◽  
Vol 18 (11) ◽  
pp. 5833
Author(s):  
Giuseppa Visalli ◽  
Alessio Facciolà ◽  
Marianna Pruiti Ciarello ◽  
Giuseppe De Marco ◽  
Maria Maisano ◽  
...  
Keyword(s):  
Chronic Exposure ◽  
Prolonged Exposure ◽  
Intestinal Inflammation ◽  
In Vitro Study ◽  
Intestinal Cell ◽  
Intestinal Cells ◽  
Barrier Dysfunction ◽  
Intestinal Cell Line

Due to ingestion of contaminated foods, the human gastrointestinal tract is the most likely site of exposure to microplastics (MPs) with gut barrier dysfunction and intestinal inflammation. Aimed to assess the effects induced by MPs with different granulometry (polystyrene (PS) 3 and 10 µm), we performed an in vitro study by using the human intestinal cell line HT29. As a novelty, we assessed the sub-chronic exposure extending the treatment up to 48 days simulating the in vivo situation. In the range of 100–1600 particles mL−1, both the PS suspensions had moderate cytotoxicity after 24 h with percentages of mortality between 6.7 and 21.6 for the 10 µm and 6.1 and 29.6 for the 3 µm PS. Microscopic observation highlighted a more pronounced lysosomal membrane permeabilization in HT29 exposed to PS 3µm. Reactive oxygen species production was higher in cells exposed to PS 10 µm, but sub-chronic exposure highlighted the ability of the cells to partially neutralize this effect. Comet-assay confirmed the temporary oxidative damage that was PS-induced. Overall, considering the very fast turnover of intestinal cells, the increase in cell mortality, equal to 25% and 11% for 3 and 10 µm PS-MPs for each time point, could trigger intestinal disorders due to prolonged exposure.

Production of immune response mediators by HT-29 intestinal cell-lines in the presence of Bifidobacterium-treated infant microbiota

2015 ◽  
Vol 6 (4) ◽  
pp. 543-552 ◽  
Author(s):  
S. Arboleya ◽  
B. Bahrami ◽  
S. Macfarlane ◽  
M. Gueimonde ◽  
G.T. Macfarlane ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune System ◽  
Intestinal Microbiota ◽  
Short Chain Fatty Acids ◽  
Model Systems ◽  
Intestinal Cell ◽  
Intestinal Cells ◽  
Suitable Model ◽  
Ht 29

The colonisation and establishment of the intestinal microbiota starts immediately at birth and is essential for the development of the intestine and the immune system. This microbial community gradually increases in number and diversity until the age of two or three years when it becomes a stable ecosystem resembling that of adults. This period constitutes a unique window of opportunity to modulate it through probiotic action, with a potential impact in later health. In the present work we have investigated how putative bifidobacterial probiotics modify the metabolic profiles and immune-modulatory properties of faecal microbiotas. An in vitro pH-controlled single-stage continuous-culture system (CCS) inoculated with infant faeces was employed to characterise the effects of two Bifidobacterium species on the intestinal microbiotas in three children, together with the effects of these modified microbiotas on cytokine production by HT-29 cells. Intestinal bacterial communities, production of short-chain fatty acids and lactate were determined by quantitative PCR and gas chromatography, respectively. Cytokines production by HT-29 cells was measured by ELISA. The combination of CCS with infant faeces and human intestinal cells provided a suitable model to evaluate the specific modulation of the intestinal microbiota and immune system by probiotics. In the CCS, infant faecal microbiotas were influenced by the addition of bifidobacteria, resulting in changes in their ability to induce the production of immune mediators by HT-29 cells. The different metabolic and immunological responses induced by the bifidobacterial species tested indicate the need to assess potential probiotics in model systems including complex intestinal microbiotas. Potential probiotic bifidobacteria can modulate the infant microbiota and its ability to induce the production of mediators of the immune response by intestinal cells.

scholarly journals Paramylon extracted from Euglena gracilis EOD-1 augmented the expression of SIRT1

2021 ◽  
Author(s):  
Hiromi Ieiri ◽  
Natsuki Kameda ◽  
Junko Naito ◽  
Takanori Kawano ◽  
Norihisa Nishida ◽  
...  
Keyword(s):  
Cell Line ◽  
Microarray Analysis ◽  
Euglena Gracilis ◽  
Epidermal Cells ◽  
Intestinal Cell ◽  
Intestinal Cells ◽  
Target Cells ◽  
Intestinal Cell Line ◽  
Human Intestinal Cell Line

AbstractEuglena gracilis, a type of microalgae, contains several nutrients and accumulates paramylon, a β-1,3-glucan. In recent studies, paramylon has shown to exhibit various activities including immunomoduratory and hepatoprotective effects. In the present study, using an in vitro cell culture system, we aimed to determine whether paramylon derived from the E. gracilis EOD-1 strain, which produces large amounts of paramylon, can augment SIRT1 expression in epidermal cells via activating gut–skin interactions. Results showed that paramylon augmented the expression of SIRT1 in Caco-2 cells, a human intestinal cell line. Furthermore, microarray analysis of Caco-2 cells treated with paramylon showed that paramylon activates epidermal cells through inducing the secretion of factors from intestinal cells. Then, we focused on skin cells as target cells of paramylon-activated intestinal cells. Results showed that secretory factors from Caco-2 cells treated with paramylon augmented the expression of SIRT1 in HaCaT cells, a human keratinocyte cell line, and that expression level of genes related to the growth and maintenance of epidermal cells were significantly changed in Caco-2 cells treated with paramylon as evidenced by microarray analysis. All these results suggest that paramylon can activate epidermal cells by inducing the production of secretory factors from intestinal cells.

scholarly journals Bifidobacterium longum subsp. infantis CECT7210 (B. infantis IM-1®) Displays In Vitro Activity against Some Intestinal Pathogens

Nutrients ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3259
Author(s):  
Lorena Ruiz ◽  
Ana Belén Flórez ◽  
Borja Sánchez ◽  
José Antonio Moreno-Muñoz ◽  
Maria Rodriguez-Palmero ◽  
...  
Keyword(s):  
Infant Health ◽  
Bifidobacterium Longum ◽  
Probiotic Strain ◽  
Intestinal Cell ◽  
Human Gastrointestinal Tract ◽  
E Coli ◽  
Intestinal Cell Line ◽  
Intestinal Pathogens ◽  
Health Promoting

Certain non-digestible oligosaccharides (NDO) are specifically fermented by bifidobacteria along the human gastrointestinal tract, selectively favoring their growth and the production of health-promoting metabolites. In the present study, the ability of the probiotic strain Bifidobacterium longum subsp. infantis CECT7210 (herein referred to as B. infantis IM-1®) to utilize a large range of oligosaccharides, or a mixture of oligosaccharides, was investigated. The strain was able to utilize all prebiotics screened. However, galactooligosaccharides (GOS), and GOS-containing mixtures, effectively increased its growth to a higher extent than the other prebiotics. The best synbiotic combination was used to examine the antimicrobial activity against Escherichia coli, Cronobacter sakazakii, Listeria monocytogenes and Clostridium difficile in co-culture experiments. C. difficile was inhibited by the synbiotic, but it failed to inhibit E. coli. Moreover, Cr. sakazakii growth decreased during co-culture with B. infantis IM-1®. Furthermore, adhesion experiments using the intestinal cell line HT29 showed that the strain IM-1® was able to displace some pathogens from the enterocyte layer, especially Cr. sakazakii and Salmonella enterica, and prevented the adhesion of Cr. sakazakii and Shigella sonnei. In conclusion, a new synbiotic (probiotic strain B. infantis IM-1® and GOS) appears to be a potential effective supplement for maintaining infant health. However, further studies are needed to go more deeply into the mechanisms that allow B.infantis IM-1® to compete with enteropathogens.

Soluble Epoxide Hydrolase as an Important Player in Intestinal Cell Differentiation

2020 ◽  
Vol 209 (4-6) ◽  
pp. 177-188
Author(s):  
Katerina Cizkova ◽  
Katerina Koubova ◽  
Tereza Foltynkova ◽  
Jana Jiravova ◽  
Zdenek Tauber
Keyword(s):  
Cell Differentiation ◽  
Colorectal Carcinoma ◽  
Cell Lines ◽  
Epoxide Hydrolase ◽  
Soluble Epoxide Hydrolase ◽  
Intestinal Cell ◽  
Caco2 Cells ◽  
Important Player ◽  
Ht 29

There is growing evidence that soluble epoxide hydrolase (sEH) may play a role in cell differentiation. sEH metabolizes biologically highly active and generally cytoprotective epoxyeicosatrienoic acids (EETs), generated from arachidonic acid metabolism by CYP epoxygenases (CYP2C and CYP2J subfamilies), to less active corresponding diols. We investigated the effect of sEH inhibitor (TPPU) on the expression of villin, CYP2C8, CYP2C9, CYP2J2, and sEH in undifferentiated and in vitro differentiated HT-29 and Caco2 cell lines. The administration of 10 μM TPPU on differentiated HT-29 and Caco2 cells resulted in a significant decrease in expression of villin, a marker for intestinal cell differentiation. It was accompanied by a disruption of the brush border when microvilli appeared sparse and short in atomic force microscope scans of HT-29 cells. Although inhibition of sEH in differentiated HT-29 and Caco2 cells led to an increase in sEH expression in both cell lines, this treatment had an opposite effect on CYP2J2 expression in HT-29 and Caco2 cells. In addition, tissue samples of colorectal carcinoma and adjacent normal tissues from 45 patients were immunostained for sEH and villin. We detected a significant decrease in the expression of both proteins in colorectal carcinoma in comparison to adjacent normal tissue, and the decrease in both sEH and villin expression revealed a moderate positive association. Taken together, our results showed that sEH is an important player in intestinal cell differentiation.

scholarly journals “Masato de Yuca” and “Chicha de Siete Semillas” Two Traditional Vegetable Fermented Beverages from Peru as Source for the Isolation of Potential Probiotic Bacteria

2021 ◽  
Author(s):  
Teresa D. Rebaza-Cardenas ◽  
Kenneth Silva-Cajaleón ◽  
Carlos Sabater ◽  
Susana Delgado ◽  
Nilda D. Montes-Villanueva ◽  
...  
Keyword(s):  
Gastrointestinal Transit ◽  
Probiotic Strain ◽  
In Vitro Digestion ◽  
Intestinal Cell ◽  
Adhesion Ability ◽  
Intestinal Cell Line ◽  
Carbohydrate Fermentation ◽  
Traditional Vegetable ◽  
Genetic Profiles

AbstractIn this work, two Peruvian beverages “Masato de Yuca,” typical of the Amazonian communities made from cassava (Manihot esculenta), and “Chicha de Siete Semillas,” made from different cereal, pseudo-cereal, and legume flours, were explored for the isolation of lactic acid bacteria after obtaining the permission of local authorities following Nagoya protocol. From an initial number of 33 isolates, 16 strains with different RAPD- and REP-PCR genetic profiles were obtained. In Chicha, all strains were Lactiplantibacillus plantarum (formerly Lactobacillus plantarum), whereas in Masato, in addition to this species, Limosilactobacillus fermentum (formerly Lactobacillus fermentum), Pediococcus acidilactici, and Weissella confusa were also identified. Correlation analysis carried out with their carbohydrate fermentation patterns and enzymatic profiles allowed a clustering of the lactobacilli separated from the other genera. Finally, the 16 strains were submitted to a static in vitro digestion (INFOGEST model) that simulated the gastrointestinal transit. Besides, their ability to adhere to the human epithelial intestinal cell line HT29 was also determined. Following both procedures, the best probiotic candidate was Lac. plantarum Ch13, a robust strain able to better face the challenging conditions of the gastrointestinal tract and showing higher adhesion ability to the intestinal epithelium in comparison with the commercial probiotic strain 299v. In order to characterize its benefit for human health, this Ch13 strain will be deeply studied in further works.

scholarly journals Gluten-induced RNA methylation changes regulate intestinal inflammation via allele-specific XPO1 translation in epithelial cells

Gut ◽  
2021 ◽  
pp. gutjnl-2020-322566
Author(s):  
Ane Olazagoitia-Garmendia ◽  
Linda Zhang ◽  
Paula Mera ◽  
Julie K Godbout ◽  
Maialen Sebastian-DelaCruz ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Autoimmune Disorder ◽  
Intestinal Cell ◽  
In Vivo Models ◽  
Knock Out ◽  
New Therapeutic Approaches ◽  
Intestinal Cell Line ◽  
Environment Characteristic

ObjectivesCoeliac disease (CD) is a complex autoimmune disorder that develops in genetically susceptible individuals. Dietary gluten triggers an immune response for which the only available treatment so far is a strict, lifelong gluten free diet. Human leucocyte antigen (HLA) genes and several non-HLA regions have been associated with the genetic susceptibility to CD, but their role in the pathogenesis of the disease is still essentially unknown, making it complicated to develop much needed non-dietary treatments. Here, we describe the functional involvement of a CD-associated single-nucleotide polymorphism (SNP) located in the 5’UTR of XPO1 in the inflammatory environment characteristic of the coeliac intestinal epithelium.DesignThe function of the CD-associated SNP was investigated using an intestinal cell line heterozygous for the SNP, N6-methyladenosine (m6A)-related knock-out and HLA-DQ2 mice, and human samples from patients with CD.ResultsIndividuals harbouring the risk allele had higher m6A methylation in the 5’UTR of XPO1 RNA, rendering greater XPO1 protein amounts that led to downstream nuclear factor kappa B (NFkB) activity and subsequent inflammation. Furthermore, gluten exposure increased overall m6A methylation in humans as well as in in vitro and in vivo models.ConclusionWe identify a novel m6A-XPO1-NFkB pathway that is activated in CD patients. The findings will prompt the development of new therapeutic approaches directed at m6A proteins and XPO1, a target under evaluation for the treatment of intestinal disorders.

scholarly journals Effect of Helicobacter pylori on Polymorphonuclear Leukocyte Migration across Polarized T84 Epithelial Cell Monolayers: Role of Vacuolating Toxin VacA and cagPathogenicity Island

2000 ◽  
Vol 68 (9) ◽  
pp. 5225-5233 ◽  
Author(s):  
Véronique Hofman ◽  
Vittorio Ricci ◽  
Antoine Galmiche ◽  
Patrick Brest ◽  
Patrick Auberger ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Polymorphonuclear Leukocyte ◽  
Broth Culture ◽  
Intestinal Cell ◽  
T84 Cells ◽  
Vacuolating Cytotoxin ◽  
Intestinal Cell Line ◽  
H Pylori

ABSTRACT Helicobacter pylori infection can induce polymorphonuclear leukocyte (PMNL) infiltration of the gastric mucosa, which characterizes acute chronic gastritis. The mechanisms underlying this process are poorly documented. The lack of an in vitro model has considerably impaired the study of transepithelial migration of PMNL induced by H. pylori. In the present work, we used confluent polarized monolayers of the human intestinal cell line T84 grown on permeable filters to analyze the epithelial PMNL response induced by broth culture filtrates (BCFs) and bacterial suspensions from different strains of H. pylori. We have evaluated the role of the vacuolating cytotoxin VacA and of the cagpathogenicity island (PAI) of H. pylori in PMNL migration via their effects on T84 epithelial cells. We noted no difference in the rates of PMNL transepithelial migration after epithelial preincubation with bacterial suspensions or with BCFs of VacA-negative or VacA-positive H. pylori strains. In contrast, PMNL transepithelial migration was induced after incubation of the T84 cells with cag PAI-positive and cagE-positiveH. pylori strains. Finally, PMNL migration was correlated with a basolateral secretion of interleukin-8 by T84 cells, thus creating a subepithelial chemotactic gradient for PMNL. These data provide evidence that the vacuolating cytotoxin VacA is not involved in PMNL transepithelial migration and that the cag PAI, with a pivotal role for the cagE gene, provokes a transcellular signal across T84 monolayers, inducing a subepithelial PMNL response.

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