Exposure to Anisakis extracts can induce inflammation on in vitro cultured human colonic cells

2017 ◽  
Vol 116 (9) ◽  
pp. 2471-2477 ◽  
Author(s):  
Antonio Speciale ◽  
Domenico Trombetta ◽  
Antonella Saija ◽  
Antonio Panebianco ◽  
Filippo Giarratana ◽  
...  
Keyword(s):  
Colonic Cells

scholarly journals Heterogeneous expression of Pil3 pilus is critical for Streptococcus gallolyticus translocation across polarized colonic epithelial monolayers

2019 ◽  
Author(s):  
Mariana Martins ◽  
Laurence du Merle ◽  
Patrick Trieu-Cuot ◽  
Shaynoor Dramsi
Keyword(s):  
Intestinal Barrier ◽  
Opportunistic Pathogen ◽  
Oral Route ◽  
Elderly Persons ◽  
Dependent Manner ◽  
Bacterial Surface ◽  
Streptococcus Gallolyticus ◽  
Heterogeneous Expression ◽  
Colonic Cells

ABSTRACTStreptococcus gallolyticus subspecies gallolyticus (Sgg) is an opportunistic pathogen responsible for septicaemia and endocarditis in elderly persons. Sgg is also a commensal of the human gastrointestinal tract. Here we demonstrate that Sgg strain UCN34 translocates across tight intestinal barriers in vitro in a Pil3-dependent manner. Confocal images of UCN34 passage across human colonic cells reveals that Sgg utilizes a paracellular pathway. Pil3 was previously shown to be expressed heterogeneously and WT UCN34 consists of about 90% of Pil3low and 10% of Pil3high cells. We found that both the Δpil3 mutant and the Pil3+ overexpressing variant could not translocate across Caco-2 and T84 barriers. Interestingly, combining live Δpil3 mutant cells with fixed Pil3+ variants in a 10:1 ratio (mimicking UCN34 WT population) allowed efficient translocation of the Δpil3 mutant. These experiments demonstrate that heterogeneous expression of Pil3 plays a key role in optimal translocation of Sgg across the intestinal barrier.ABSTRACT IMPORTANCEStreptococcus gallolyticus subsp. gallolyticus (Sgg) is an opportunistic pathogen responsible for septicemia and infective endocarditis in elderly persons. Sgg is a commensal of the rumen of herbivores and transmission to humans most probably occurs through the oral route. In this work, we have studied how this bacterium crosses the intestinal barrier using well-known in vitro models. Confocal microscopy images revealed that Sgg UCN34 can traverse the epithelial monolayer in between adjacent cells. We next showed that passage of Sgg from the apical to the basolateral compartment is dependent on the heterogenous expression of the Pil3 pilus at the bacterial surface. We hypothesize that Pil3high cocci adhere firmly to epithelial cells to activate transient opening of tight junctions thereby allowing the traversal of Pil3low bacteria.

scholarly journals Colonic Epithelial PHLPP2 Deficiency Promotes Colonic Epithelial Pyroptosis by Activating the NF-κB Signaling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
De-feng Li ◽  
Xin Chang ◽  
Jiu-long Zhao ◽  
Xuan-min Chen ◽  
Zheng-lei Xu ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
Chronic Inflammatory Disease ◽  
Pleckstrin Homology Domain ◽  
Caspase 1 ◽  
Barrier Defect ◽  
Protein Phosphatase 2 ◽  
Inflammatory Caspases ◽  
Colonic Cells

Background. Ulcerative colitis (UC) is a chronic inflammatory disease with increasing prevalence worldwide. Barrier defect in intestinal epithelial cells (IECs) is one of the main pathogeneses in UC. Pyroptosis is a programmed lytic cell death and is triggered by inflammatory caspases, while little is known about its role in UC. Methods. Differentially expressed genes (DEGs) were identified by comparing UC patients with healthy controls from the GEO datasets. The candidate genes involved in pyroptosis were obtained, and the underlying molecular mechanism in the progression of UC was explored in vivo and in vitro. Results. Pleckstrin homology domain leucine-rich repeat protein phosphatase 2 (PHLPP2), a protein phosphatase, was downregulated and involved in regulating inflammation-induced IEC pyroptosis by modulating the NF-κB signaling in UC through bioinformatics analysis. Moreover, we demonstrated that PHLPP2 was downregulated in UC patients and UC mice. Besides, we found that PHLPP2 depletion activated the NF-κB signaling and increased the expressions of caspase-1 P20, Gasdermin N, IL-18, and IL-1β contributing to IEC pyroptosis and inflammation in UC mice. Furthermore, we found that PHLPP2-/- mice developed hypersensitivity to dextran sulfate sodium (DSS) treatment toward colitis showing activated NF-κB signaling and dramatically induced expressions of caspase-1 P20, Gasdermin N, IL-18, and IL-1β. Mechanically, this inflammation-induced downregulation of PHLPP2 was alleviated by an NF-κB signaling inhibitor in intestinal organoids of PHLPP2-/- mice and fetal colonic cells. Conclusions. PHLPP2 downexpression activated the NF-κB signaling and promoted the IEC pyroptosis, leading to UC progression. Therefore, PHLPP2 might be an attractive candidate therapeutic target for UC.

scholarly journals Time-dependent activity and expression of glutathione S-transferases in the human colon adenocarcinoma cell line Caco-2

1989 ◽  
Vol 264 (2) ◽  
pp. 613-616 ◽  
Author(s):  
W H N Peters ◽  
H M J Roelofs
Keyword(s):  
Cell Line ◽  
Glutathione Transferase ◽  
Colon Adenocarcinoma ◽  
Human Colon ◽  
Glutathione S Transferase ◽  
Human Colon Carcinoma Cell ◽  
Cells In Culture ◽  
Glutathione S Transferases ◽  
Colonic Cells

The human colon carcinoma cell line Caco-2 was examined for glutathione S-transferase (GST) composition and activity. Freshly seeded cells and cells until 4 days after confluency contain only the placental (Pi) form of glutathione transferase. Cells in culture for longer periods start to express class-Alpha GST isoenzymes. Confluent cells in culture for 20 days or longer contain up to 90% class-Alpha GST. Class-Mu GSTs are not detectable. GST activity gradually increases from 564 +/- 28 to 5381 +/- 165 nmol/min per mg of protein at day 0 and 32 after confluency respectively. With regard to GST composition, Caco-2 cells in culture for longer periods most resemble small-intestinal cells, whereas short-time cultures have characteristics of colonic cells. This cell line is very well suited for the study of both the in vitro properties and the expression of class Alpha and Pi GSTs.

Riboflavin uptake by human-derived colonic epithelial NCM460 cells

2000 ◽  
Vol 278 (2) ◽  
pp. C270-C276 ◽  
Author(s):  
Hamid M. Said ◽  
Alvaro Ortiz ◽  
Mary Pat Moyer ◽  
Norimoto Yanagawa
Keyword(s):  
Epithelial Cells ◽  
Large Intestine ◽  
Water Soluble ◽  
Cellular Regulation ◽  
Colonic Epithelial Cells ◽  
Normal Microflora ◽  
Intracellular Ca ◽  
Energy Dependent ◽  
Colonic Cells

Normal microflora of the large intestine synthesize a number of water-soluble vitamins including riboflavin (RF). Recent studies have shown that colonic epithelial cells posses an efficient carrier-mediated mechanism for absorbing some of these micronutrients. The aim of the present study was to determine whether colonic cells also posses a carrier-mediated mechanism for RF uptake and, if so, to characterize this mechanism and study its cellular regulation. Confluent monolayers of the human-derived nontransformed colonic epithelial cells NCM460 and [3H]RF were used in the study. Uptake of RF was found to be 1) appreciable and temperature and energy dependent; 2) Na+ independent; 3) saturable as a function of concentration with an apparent K mof 0.14 μM and V max of 3.29 pmol ⋅ mg protein−1 ⋅ 3 min−1; 4) inhibited by the structural analogs lumiflavin and lumichrome ( K i of 1.8 and 14.1 μM, respectively) but not by the unrelated biotin; 5) inhibited in a competitive manner by the membrane transport inhibitor amiloride ( K i = 0.86 mM) but not by furosemide, DIDS, or probenecid; 6) adaptively regulated by extracellular RF levels with a significant and specific upregulation and downregulation in RF uptake in RF-deficient and oversupplemented conditions, respectively; and 7) modulated by an intracellular Ca2+/calmodulin-mediated pathway. These studies demonstrate for the first time the existence of a specialized carrier-mediated mechanism for RF uptake in an in vitro cellular model system of human colonocytes. This mechanism appears to be regulated by extracellular substrate level and by an intracellular Ca2+/calmodulin-mediated pathway. It is suggested that the identified transport system may be involved in the absorption of bacterially synthesized RF in the large intestine and that this source of RF may contribute toward RF homeostasis, especially that of colonocytes.

scholarly journals In Vitro Evaluation of the Effects of Commercial Prebiotic GOS and FOS Products on Human Colonic Caco–2 Cells

Nutrients ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1281
Author(s):  
Geraldine M. Flaujac Lafontaine ◽  
Neville M. Fish ◽  
Ian F. Connerton
Keyword(s):  
Animal Feed ◽  
Feed Additives ◽  
Differentially Expressed ◽  
Control Group ◽  
Intestinal Epithelial ◽  
Beneficial Effects ◽  
Prebiotic Oligosaccharides ◽  
Energy Dependent ◽  
Colonic Cells

Prebiotic oligosaccharides are widely used as human and animal feed additives for their beneficial effects on the gut microbiota. However, there are limited data to assess the direct effect of such functional foods on the transcriptome of intestinal epithelial cells. The purpose of this study is to describe the differential transcriptomes and cellular pathways of colonic cells directly exposed to galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS). We have examined the differential gene expression of polarized Caco–2 cells treated with GOS or FOS products and their respective mock-treated cells using mRNA sequencing (RNA-seq). A total of 89 significant differentially expressed genes were identified between GOS and mock-treated groups. For FOS treatment, a reduced number of 12 significant genes were observed to be differentially expressed relative to the control group. KEGG and gene ontology functional analysis revealed that genes up-regulated in the presence of GOS were involved in digestion and absorption processes, fatty acids and steroids metabolism, potential antimicrobial proteins, energy-dependent and -independent transmembrane trafficking of solutes and amino acids. Using our data, we have established complementary non-prebiotic modes of action for these frequently used dietary fibers.

scholarly journals In Vitro Evaluation of the Effects of Prebiotics GOS and FOS on Human Colonic Caco–2 Cells

2020 ◽  
Author(s):  
Geraldine Lafontaine ◽  
Neville Fish ◽  
Ian Connerton
Keyword(s):  
Animal Feed ◽  
Feed Additives ◽  
Differentially Expressed ◽  
Control Group ◽  
Intestinal Epithelial ◽  
Beneficial Effects ◽  
Prebiotic Oligosaccharides ◽  
Energy Dependent ◽  
Colonic Cells

Prebiotic oligosaccharides are widely used as human and animal feed additives for their beneficial effects on the gut microbiota. However, there are limited data to assess the direct effect of such functional foods on the transcriptome of intestinal epithelial cells. The purpose of this study is to describe the differential transcriptomes and cellular pathways of colonic cells directly exposed to galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS). We have examined the differential gene expression of polarized Caco-2 cells treated with GOS or FOS and their respective mock-treated cells using mRNA sequencing (RNA-seq). A total of 89 significant differentially expressed genes were identified between GOS and mock-treated groups. For FOS treatment, a reduced number of 12 significant genes were observed to be differentially expressed relative to the control group. KEGG and Gene Ontology functional analysis revealed that genes up-regulated in the presence of GOS were involved in digestion and absorption processes, fatty acids and steroids metabolism, potential antimicrobial proteins, energy-dependent and -independent transmembrane trafficking of solutes and amino acids. Using our data, we have established complementary non-prebiotic modes of action for these frequently used dietary fibers.

Downregulation of a human colonic sialyltransferase by a secondary bile acid and a phorbol ester

1998 ◽  
Vol 274 (3) ◽  
pp. G599-G606 ◽  
Author(s):  
Ming Li ◽  
Ravi Vemulapalli ◽  
Asad Ullah ◽  
Leighton Izu ◽  
Michael E. Duffey ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Phorbol Ester ◽  
Cancer Cell Line ◽  
Colon Cancer Cell Line ◽  
Mrna Levels ◽  
Membrane Glycoproteins ◽  
Secondary Bile Acid ◽  
Colonic Cells

Fecal constituents such as bile acids and increased sialylation of membrane glycoproteins by α-2,6-sialyltransferase (HST6N-1) may contribute to colorectal tumorigenesis. We hypothesized that bile acids and phorbol ester [12- O-tetradecanoylphorbol-13-acetate (TPA)] would upregulate HST6N-1 in colonic cells. However, deoxycholate (DOC) (300 μmol/l), a secondary bile acid, and TPA (20 ng/ml) decreased expression of an ∼100-kDa glycoprotein bearing α-2,6-linked sialic acid in a colon cancer cell line (T84) in vitro. HST6N-1 mRNA levels were reduced ∼80% by treatment (≤24 h) with DOC or TPA but not by cholate, a primary bile acid. Treatment (24 h) with DOC or TPA decreased activity of this enzyme to 30% and 13% of control, respectively. These effects of DOC and TPA were transcriptional and were mediated by Ca2+ and protein kinase C, respectively. Thus DOC and TPA both downregulated, and did not upregulate, α-2,6-sialyltransferase expression in vitro, but by different transduction pathways. As colorectal tumors grow, their progressive removal from the fecal milieu that normally downregulates this enzyme may favor invasion and metastasis.

scholarly journals Indole-3-Carbinol Inhibits Citrobacter rodentium Infection through Multiple Pathways Including Reduction of Bacterial Adhesion and Enhancement of Cytotoxic T Cell Activity

Nutrients ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 917
Author(s):  
Yanbei Wu ◽  
Qiang He ◽  
Liangli Yu ◽  
Quynhchi Pham ◽  
Lumei Cheung ◽  
...  
Keyword(s):  
Colon Cancer ◽  
T Cell ◽  
Intestinal Inflammation ◽  
Control Diet ◽  
Cell Activity ◽  
Fecal Excretion ◽  
Cytotoxic T Cell ◽  
Citrobacter Rodentium ◽  
Colonic Cells

Intestinal inflammation is associated with an increased risk of developing colorectal cancer and may result from dysregulated responses to commensal bacteria or exposure to bacterial pathogens. Dietary modulation of intestinal inflammation may protect against development of colon cancer. However, the precise diet-derived components and underlying mechanisms remain elusive. Citrobacter rodentium (Cr) induces acute intestinal inflammation and has been used to study the role of inflammation in the susceptibility to colon cancer. Here we examine the effects of indole-3-carbinol (I3C), a dietary compound with anticarcinogenic properties, on intestinal immune and inflammatory responses to Cr infection and adhesion to colonic cells in vitro. C57BL/6J mice were fed a diet with/without 1 μmol/g I3C and infected with Cr. Compared to infected mice fed with a control diet, consumption of a 1 μmol I3C/g diet significantly reduced fecal excretion of Cr, Cr colonization of the colon, and reduced colon crypt hyperplasia. Furthermore, expression of Cr-induced inflammatory markers such as IL-17A, IL-6, and IL1β were attenuated in infected mice fed with the I3C diet, compared to mice fed a control diet. The expression of cytotoxic T cell markers CD8 and FasL mRNA were increased in I3C-fed infected mice. In-vitro, I3C inhibited Cr growth and adhesion to Caco-2 cells. I3C alleviates Cr-induced murine colitis through multiple mechanisms including inhibition of Cr growth and adhesion to colonic cells in vitro and enhancement of cytotoxic T cell activity.

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