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Gene Regulation of Intestinal Porcine Epithelial Cells IPECM-bM-^@M-^QJ2 is Dependent on the Site of Deoxynivalenol Toxicological Action

ABSTRACT: Here we analysed different mechanisms of apical and basolateral deoxynivalenol (DON) toxicity reflected in the gene expression. We used the jejunum derived, polarized intestinal porcine epithelial cell line IPECM-bM-^@M-^QJ2 as an in vitro cell culture model. Luminal and blood stream DON intoxication was mimicked by a DON application from the apical or basolateral compartment of ThinCertM-BM-. membrane inserts for 72M-BM- h. We compared the genome wide gene expression of untreated and DON-treated IPEC-J2 cells by the GeneChipM-BM-. Porcine Genome Array of Affymetrix.

ORGANISM(S): Sus scrofa

SUBMITTER: Klaus Wimmers

PROVIDER: E-GEOD-33246 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene Regulation of Intestinal Porcine Epithelial Cells IPEC‑J2 is Dependent on the Site of Deoxynivalenol Toxicological Action

Project description:Here we analysed different mechanisms of apical and basolateral deoxynivalenol (DON) toxicity reflected in the gene expression. We used the jejunum derived, polarized intestinal porcine epithelial cell line IPEC‑J2 as an in vitro cell culture model. Luminal and blood stream DON intoxication was mimicked by a DON application from the apical or basolateral compartment of ThinCert® membrane inserts for 72 h.

2012-12-31 | GSE33246 | GEO

Neisseria meningitidis elicits a pro-inflammatory response involving IM-NM-:BM-NM-6 in a human blood-cerebrospinal fluid barrier model.

Project description:The human-specific, Gram-negative bacterium Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis world-wide. It has been described that Nm can enter the central nervous system via the blood-cerebrospinal fluid barrier (BCSFB), which is constituted by the epithelial cells of the choroid plexus. Using a recently established in vitro model of the human BCSFB based on human malignant choroid plexus papilloma (HIBCPP) cells we investigated the cellular response of HIBCPP cells challenged with the meningitis-causing Nm strain MC58. In comparison we analysed the answer to the closely related unencapsulated carrier isolate Nm M-NM-114. Transcriptome analysis revealed a stronger transcriptional response after infection with strain MC58, in particular with its capsule deficient mutant MC58siaD-, which correlated with bacterial invasion levels. Expression evaluation and Gene Set Enrichment Analysis pointed to a NF-M-NM-:B-mediated pro-inflammatory immune response involving up-regulation of the transcription factor IM-NM-:BM-NM-6. Consistent with this, infected cells secreted significant levels of pro-inflammatory chemokines and cytokines, among others, IL8, CXCL1-3 and the IM-NM-:BM-NM-6 target gene product IL6. Expression profile of pattern recognition receptors in HIBCPP cells and the response to specific agonists indicates that TLR2 rather than TLR4 is involved in the cellular reaction following Nm infection. Human malignant choroid plexus papilloma (HIBCPP) cells were infected from the basolateral side with the meningitis-causing Neisseria meningitidis disease isolate MC58, its non-capsulated mutant MC58siaD- and the Neisseria meningitidis carrier isolate M-NM-114 for 4 h.The transcriptional response of HIBCPP cells to the different Neisseria meningitidis strains was evaluated by microarray analysis. Untreated HIBCPP cells served as control. Three replicates of each condition were analysed.

2014-09-16 | E-GEOD-42870 | biostudies-arrayexpress

Impact of biomechanical stimulation in the toxicity of deoxynivalenol in A431 cells: impairment of “dynamic” cell functions

Project description:Morphology together with the capability to respond to surrounding stimuli are the key elements governing the spatial interaction of a living cell with the environment. In this respect, biomechanical stimulation can trigger significant physiological cascades that can potentially modulate or interact with toxicity. Deoxynivalenol (DON, vomitoxin) is one of the most prevalent mycotoxins produced by Fusarium spp. and it was used to explore the delicate interaction between biomechanical stimulation and cytotoxic processes in A431 cells. In fact, in addition of being a well-known food contaminant, DON is becoming a relevant toxin also for other organ systems. The combination between biomechanical stimulation and the mycotoxin revealed how DON can impair crucial functions like cellular morphology and lysosome trafficking at concentrations far below those known to be cytotoxic in routine toxicity studies. Moreover, sub-toxic concentrations of DON (0.1-1 μM) impaired the capability of A431 cells to respond to a biomechanical stimulation that normally sustains trophic effects in this cell line. Ultimately, the effects of DON (0.1-10 μM) were partially modulated by the application of uniaxial stretching (0.5Hz, 24h, 15% deformation) suggesting a deep cross-talk between the cytotoxic potential of the mycotoxin and the biomechanical stimulation.

2018-07-31 | PXD008996 | Pride

Deoxynivalenol and A431 epidermoid squamous carcinoma cells: proteomic profiling reveals the pathways connecting ribosomal inhibition with the loss of function of the cell membrane

Project description:The possibility to combine proteomics workflows to classical experimental approaches is continuously opening new insights in all branches of biological sciences. Deoxynivalenol (vomitoxin, DON) is a secondary metabolite produced by Fusarium spp. fungi and it is one of the most recurrent mycotoxins worldwide. DON is known to inhibit protein synthesis and as such interact with the cell models in multiple and complex ways whose comprehension enormously benefit of the system toxicology approach. For the purpose of this study epidermoid squamous cell carcinoma cells A431 were incubated with DON for 24h and toxin dependent variation of the proteome profile was obtained with a LC-MS/MS approach using a reversed phase nanoLC-System (Ultimate 3000RSLC Thermo Fisher Scientific, Austria) hyphenated to a High Resolution Orbitrap Mass Spectrometer (Thermo Fisher Scientific™ Q Exactive™ Classic). DON significantly down-regulated ribosomal proteins, as well as mitochondrial respiratory chain elements and transport proteins (TOMM22; TOMM40; TOMM70A). In line with the loss of mitochondrial function, altered metabolic capability was observed, with particular impairment of lipid synthesis cascade. Effect of the mycotoxin on cell membrane was verified by confocal microscopy (morphology) and by membrane fluidity measures (biophysical properties).

2021-04-05 | PXD011474 | Pride

Influence of Deoxynivalenol-contaminated diet on the liver gene expression of chicken.

Project description:The mycotoxin deoxynivalenol (DON) is a secondary metabolite from Fusarium species and is frequently present on wheat and other cereals. The main effects of DON are a reduction of the feed intake and reduced weight gain of broilers. At the molecular level DON binds to the 60S ribosomal subunit and inhibits subsequently protein synthesis at the translational level. It has been suggested that cells and tissues with high protein turnover rate, like the liver and small intestine, are most affected by DON. However, little is known about other effects of DON e.g. at the transcriptional level. Therefore we decided to perform a microarray analysis, which allows us the investigation of thousands of transcripts in one experiment. The one-day old broiler chicks were separated into four groups. The diets consisted of a control diet and of three diets with moderate concentrations of 1.0, 2.5 and 5.0 mg DON/kg feed, which was attained by exchanging uncontaminated wheat with naturally DON-contaminated wheat up to the intended concentration. The chicken were held at standard conditions for 23 days and received their diet ad libitum. After slaughter the gene expression was determined in the liver of three samples per group.

2011-11-05 | E-GEOD-25185 | biostudies-arrayexpress

Comparative effect of DON and plant or microbial biotransformation of DON on gene expression of piglets jejunal explants

Project description:Deoxynivalenol (DON) is a frequent mycotoxin in grains, produced by Fusarium fungi, which demonstre multiple side effects such as modulation of immune responses, reduced feed intake and weight gain or impairment of the intestinal barrier function. Among animal species, pigs are the best model for humans and are very sensitive to DON. In wheat, DON can be conjugated to glucose to form DON-3-Î²-D-glucoside (D3G). Some bacteria isolated from digestive tracts or soil, are also able to de-epoxydize or epimerize DON to metabolites such as deepoxy-deoxynivalenol (DOM-1) or 3-epi-deoxynivalenol (epi-DON). The toxicity of these DON metabolites is poorly documented. By the way of ingestion, the intestine is the first organ exposed to these molecules and so constitute a relevant model. The aim of this study was to compare the intestinal toxicity of three DON metabolites (D3G, DOM-1 and epi-DON) with the one of DON. Intestinal explants from 6 pigs were treated with 10mM DON, D3G, DOM-1 or epi-DON for 4 hours and transcriptomic analysis was performed using an âAgilent Porcinet 60Kâ. Jejunal explants from 4 piglets aged of 5 weeks were sampled and exposed in vitro to differents molecules (DON, D3G, DOM-1 & 3-epi-DON) at 10ÂµM during 4h. Then RNA was extracted.

2016-07-20 | E-GEOD-66918 | biostudies-arrayexpress

Multiple myeloma-derived extracellular vesicles modulate the bone marrow immune microenvironment

Project description:One of the hallmarks of multiple myeloma (MM) is a permissive BM microenvironment. Increasing evidence suggest that cell-to-cell communication between myeloma and immune cells via tumor cell-derived extracellular vesicles (EV) plays a key role in the pathogenesis of MM. Hence, we aimed to explore BM immune alterations induced by MM-derived EV. For this, we inoculated immunocompetent BALB/cByJ mice with a myeloma cell line - MOPC315.BM -, inducing a MM phenotype. Upon tumor establishment, characterization of the BM microenvironment revealed the expression of both activation and suppressive markers by lymphocytes, such as Granzyme b and PD-1, respectively. In addition, conditioning of the animals with MOPC315.BM-derived EV, before transplantation of the MOPC315.BM tumor cells, did not anticipate the disease phenotype. However, it induced features of suppression in the BM milieu, such as an increase in PD-1 expression by CD4+ T cells. Overall, our findings reveal the involvement of MOPC315.BM-derived EV protein content as promoters of immune niche remodeling, strengthening the importance of assessing the mechanisms by which MM may impact the immune microenvironment.

2022-07-05 | PXD032934 | Pride

Proteomic changes in HepaRG cells exposed to mycotoxins, part 2

Project description:Proteomic changes associated with the individual and combined exposures of deoxynivalenol (DON) and zearalenone (ZEA) were investigated in human hepatocytes (HepaRG cell line) after 24 hour expousre and at low cytotoxicity levels using liquid chromatography coupled to tandem mass spectrometry.

2017-12-13 | PXD006267 | Pride

Proteomic changes in HepaRG cells exposed to mycotoxins

Project description:Proteomic changes associated with the individual and combined exposures of deoxynivalenol (DON) and zearalenone (ZEA) were investigated in human hepatocytes (HepaRG cell line) after only 1h exposure and at low cytotoxicity levels using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS)

2017-12-13 | PXD005840 | Pride

Polar infection of Echovirus 30 causes differential barrier affection and gene regulation at the blood-CSF barrier

Project description:Among the non-polio enterovirus (NPEV) Echovirus-30 (E-30) is responsible for extensive global outbreaks of meningitis in children. To gain access to the central nervous system (CNS), E-30 first have to cross one of the two main barriers, the epithelial blood-cerebrospinal fluid (CSF) barrier (BCSFB) or the endothelial blood-brain barrier (BBB). Previously it has been shown that several meningitis causing bacteria preferentially infect human choroid plexus papilloma cells (HIBCPP) in a polar fashion from the basolateral cell side. Here, we investigated the polar infection with E-30 of HIBCPP cells. Both, apical and basolateral infections caused a significant decrease of the transepithelial resistance (TEER) of HIBCPP cells in a dose-dependent manner. However, to reach the same TEER decrease the multiplicity of infection (MOI) of the apical infection had to be 20 times higher than the basolateral infection. In line with this finding the number of infected cells at respective time-points after basolateral infection was significantly higher compared to apical infection. Cytotoxic effects of E-30 on HIBCPP cells during basolateral infection were observed following prolonged infection, and appeared more drastically compared to the apical infection. Evaluation of massive analysis of cDNA ends (MACE) RNA data comparing basolateral versus apical infection, revealed a distinct pattern of up- and down-regulated genes depending on the side of infection. Especially, the down-regulation of ITGα5 upon basolateral infection can be correlated with a more drastic impact of E-30 on the HIBCPP cells when compared to apical infection, leading to structural changes. Also, type 3 interferon genes might be a factor involved in the polar effect of E-30 on HIBCPP. Altogether, the data highlights the polar effect of E-30 in HIBCPP cells as an important factor for an efficient infection.

2020-10-02 | GSE146890 | GEO

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