Neutrophil-mediated epithelial injury during transmigration: role of elastase

2001 ◽  
Vol 281 (3) ◽  
pp. G705-G717 ◽  
Author(s):  
Hedy H. Ginzberg ◽  
Vera Cherapanov ◽  
Qin Dong ◽  
Andre Cantin ◽  
Christopher A. G. McCulloch ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Barrier Function ◽  
Junctional Complex ◽  
Epithelial Barrier ◽  
Epithelial Cell Line ◽  
Leukocyte Elastase ◽  
Epithelial Barrier Function ◽  
Apical Junctional Complex ◽  
Apical Junctions ◽  
E Cadherin

Neutrophil-mediated injury to gut epithelium may lead to disruption of the epithelial barrier function with consequent organ dysfunction, but the mechanisms of this are incompletely characterized. Because the epithelial apical junctional complex, comprised of tight and adherens junctions, is responsible in part for this barrier function, we investigated the effects of neutrophil transmigration on these structures. Using a colonic epithelial cell line, we observed that neutrophils migrating across cell monolayers formed clusters that were associated with focal epithelial cell loss and the creation of circular defects within the monolayer. The loss of epithelial cells was partly attributable to neutrophil-derived proteases, likely elastase, because it was prevented by elastase inhibitors. Spatially delimited disruption of epithelial junctional complexes with focal loss of E-cadherin, β-catenin, and zonula occludens 1 was observed adjacent to clusters of transmigrating neutrophils. During neutrophil transmigration, fragments of E-cadherin were released into the apical supernatant, and inhibitors of neutrophil elastase prevented this proteolytic degradation. Addition of purified leukocyte elastase also resulted in release of E-cadherin fragments, but only after opening of tight junctions. Taken together, these data demonstrate that neutrophil-derived proteases can mediate spatially delimited disruption of epithelial apical junctions during transmigration. These processes may contribute to epithelial loss and disruption of epithelial barrier function in inflammatory diseases.

Proteolytic antigens interfere with endosome/lysosome fusion in epithelial cells

2013 ◽  
Vol 91 (6) ◽  
pp. 449-454 ◽  
Author(s):  
Yu-Wei Liao ◽  
Xing-Mao Wu ◽  
Jia Jia ◽  
Xiao-Lei Wu ◽  
Hong Tao ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Barrier Function ◽  
Epithelial Barrier ◽  
Epithelial Cell Line ◽  
Airway Epithelial ◽  
Barrier Dysfunction ◽  
Epithelial Barrier Function ◽  
Epithelial Barrier Dysfunction

The airway epithelial barrier function is important in maintaining the homeostasis in the body. A number of airway disorders are associated with the epithelial barrier dysfunction. The present study aims to elucidate a possible mechanism by which the proteolytic allergens compromise the epithelial barrier function. The airway epithelial cell line, RPMI 2650 cells (Rp cells) and kidney epithelial cell line, MDCK cells, were cultured to be monolayers and used as an in vitro epithelial barrier model. House dust mite antigen, Der P1 (Der) was used as an antigen that has the proteolytic property. The epithelial barrier permeability and transepithelial resistance (TER) were used as the indicators of epithelial barrier function. Both epithelial cell lines could endocytose Der in the culture. Some of the Der was transported across the epithelial barrier to the basal chambers of the Transwells without affecting the TER. The endocytic Der could suppress the expression of ubiquitin E3 lases A20 and further interfered with the fusion of endosome/lysosome in the epithelial cells. Mite antigen, Der, can interfere with the fusion of endosome/lysosome in epithelial cells to induce the epithelial barrier dysfunction.

scholarly journals Chemerin Regulates Epithelial Barrier Function of Mammary Glands in Dairy Cows

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3194
Author(s):  
Yutaka Suzuki ◽  
Sachi Chiba ◽  
Koki Nishihara ◽  
Keiichi Nakajima ◽  
Akihiko Hagino ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Barrier Function ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Epithelial Barrier ◽  
Epithelial Tissue ◽  
Epithelial Barrier Function

Epithelial barrier function in the mammary gland acts as a forefront of the defense mechanism against mastitis, which is widespread and a major disorder in dairy production. Chemerin is a chemoattractant protein with potent antimicrobial ability, but its role in the mammary gland remains unelucidated. The aim of this study was to determine the function of chemerin in mammary epithelial tissue of dairy cows in lactation or dry-off periods. Mammary epithelial cells produced chemerin protein, and secreted chemerin was detected in milk samples. Chemerin treatment promoted the proliferation of cultured bovine mammary epithelial cells and protected the integrity of the epithelial cell layer from hydrogen peroxide (H2O2)-induced damage. Meanwhile, chemerin levels were higher in mammary tissue with mastitis. Tumor necrosis factor alpha (TNF-α) strongly upregulated the expression of the chemerin-coding gene (RARRES2) in mammary epithelial cells. Therefore, chemerin was suggested to support mammary epithelial cell growth and epithelial barrier function and to be regulated by inflammatory stimuli. Our results may indicate chemerin as a novel therapeutic target for diseases in the bovine mammary gland.

scholarly journals Hyaluronan and Layilin Mediate Loss of Airway Epithelial Barrier Function Induced by Cigarette Smoke by Decreasing E-cadherin

2012 ◽  
Vol 287 (50) ◽  
pp. 42288-42298 ◽  
Author(s):  
Rosanna Malbran Forteza ◽  
S. Marina Casalino-Matsuda ◽  
Nieves S. Falcon ◽  
Monica Valencia Gattas ◽  
Maria E. Monzon
Keyword(s):  
Cigarette Smoke ◽  
Barrier Function ◽  
Epithelial Barrier ◽  
Airway Epithelial ◽  
Epithelial Barrier Function ◽  
E Cadherin

P150 ENHANCING EPITHELIAL BARRIER FUNCTION WITH NOVEL E-CADHERIN ACTIVATING MONOCLONAL ANTIBODIES REDUCES INFLAMMATORY BOWEL DISEASE PROGRESSION IN MOUSE MODELS

2020 ◽  
Vol 26 (Supplement_1) ◽  
pp. S29-S30
Author(s):  
Chirosree Bandyopadhyay ◽  
Leslayann Schecterson ◽  
Barry Gumbiner
Keyword(s):  
Inflammatory Bowel Disease ◽  
Monoclonal Antibodies ◽  
Treatment Group ◽  
Mouse Models ◽  
Barrier Function ◽  
Epithelial Barrier ◽  
Transfer Model ◽  
Epithelial Barrier Function ◽  
Inflammatory Bowel ◽  
E Cadherin

Abstract Deficits in gastrointestinal (GI) epithelial barrier function play important roles in the pathogenesis of Inflammatory Bowel Disease (IBD). The CDH1 gene encoding E-cadherin, a key component of the epithelial junctional complex, is associated with Ulcerative Colitis (UC), and perhaps Crohn’s disease (CD). E-cadherin is the principle adhesive component of the adherens junction, and it regulates paracellular permeability by facilitating the formation of tight junctions and organizing the entire epithelial junction complex. We have identified monoclonal antibodies (mAbs) that bind to E-cadherin and activate adhesion in a variety of epithelial cells. In this study, we aim to test the hypothesis that strengthening E-cadherin adhesion with activating mAbs will enhance barrier function and decrease progression of IBD while maintaining mucosal health and homeostasis. Mouse mAbs to E-cadherin have been tested in vivo using the IL10-knock out mouse and adoptive T cell transfer model of colitis with similar histological evaluation. Transfer of CD4+CD45Rb high T cells from donor to immunocompromised mice produced typical histologic lesions for the adoptive transfer model including inflammation of the mucosa/submucosa, crypt damage, erosions, edema, and epithelial hyperplasia. E-Cadherin activating mAb (r56.4) treatment reduced total colitis score, mucosal hyperplasia, inflammation, gland loss scores, and neutrophilic infiltration in CD45Rb high T cell recipient mice compared to control E-cad mAb (r19.1–10) treatment. In IL10 KO BL6 mouse model of colitis, average lesion severity scores were lower in the r56.4 treatment group in comparison to the r19.1–10 treatment group for all the histological hallmarks of colitis. Further studies are in progress to investigate the therapeutic potential of E-Cadherin mAbs in the rescue of inflammation in pre-clinical mouse models of colitis.

14 COLONIC EPITHELIAL CELL-SPECIFIC AFTIPHILIN KNOCKDOWN REGULATES EPITHELIAL BARRIER FUNCTION THROUGH MYOSIN LIGHT CHAIN-ASSOCIATED ACTIN ORGANIZATION IN VITRO AND INTESTINAL LENGTH IN VIVO

2020 ◽  
Vol 26 (Supplement_1) ◽  
pp. S28-S28
Author(s):  
Ivy Ka Man Law ◽  
Carl Rankin ◽  
Charalabos Pothoulakis
Keyword(s):  
Epithelial Cell ◽  
Barrier Function ◽  
Intestinal Epithelial Cell ◽  
Epithelial Barrier ◽  
Intestinal Epithelial ◽  
Knock Out ◽  
Epithelial Barrier Function ◽  
Actin Organization

Abstract Background and Aims Colonic epithelial integrity is often compromised during colonic inflammation and Inflammatory Bowel Disease. Aftiphilin (AFTPH) is a downstream target of microRNA-133a and its expression is reduced in colonic tissues of wild type mice from experimental colitis models and colonic biopsies from patients with ulcerative colitis. We have previously shown that AFTPH is involved in regulating intestinal epithelial barrier function and actin organization in human colonic epithelial cells in vitro (DDW 2016). On the other hand, our results suggested that global aftiphilin knock-out is embryonic lethal in mouse models (DDW 2019). Here, we further examined the role of AFTPH in regulating actin organization in vitro and characterize the colonic epithelial cell-specific aftiphilin knock-out mice. Methods Human colonic epithelial NCM460 cells were transfected with si-RNA against AFTPH to achieve transient AFTPH gene-silencing. Stable AFTPH knock-down clones were generated by transducing Caco2-BBE cells with recombinant lentivirus carrying sh-AFTPH or control sh-RNA. To create intestinal epithelial cell-specific aftiphilin knock-out mice, Aftph flox/flox mice were cross-bred with B6.Cg-Tg(Vil1-cre)997Gum/J mice, which express Villin-driven Cre recombinase (Vil-Cre), to generate intestinal epithelial cell-specific aftiphilin knock-out mice (Aftph Vil-/Vil-). Protein expression of F- and G-actin and p70S6K were detected using Western blot. Tissues from various organs were collected with Aftph Vil-/Vil- and its wildtype counterparts at 12 weeks. Results Results from western blot analysis showed that F-/G-actin ratio in AFTPH gene-silenced NCM460 cells were 0.6±0.17 fold, when compared to the treatment control. In addition, AFTPH gene-silencing in human colonic epithelial cells activated p70S6K, a kinase that is involved in actin organization, when compared to treatment control (1.2±0.15 vs. 2.0±0.15, p=0.0354). Furthermore, transepithelial electric resistance (TER) of Caco2-BBE cells deficient in AFTPH is significantly lower than that of control cells (0.5±0.07 fold). Lastly, in vivo intestinal epithelial cell-specific Aftph knock-out increased the length of small intestine, when compared to that of wild type mice (30.7±0.33 vs. 34.8±0.97, p=0.02), while the tissue weight of spleen to body weight was reduced (0.30±0.011 vs. 0.26±0.006, p=0.0169). Summary and Conclusions Our results indicate that AFTPH directly regulates epithelial barrier function and actin organization through mediating F-/G-actin ratio in human colonic epithelial cells, possibly through p70S6K. Importantly, intestinal epithelial cell-specific knock-out in vivo increased intestinal length and reduced size of the spleen. Our results suggested that AFTPH is crucial in regulating colonic epithelial barrier function in vitro and intestinal homeostasis.

scholarly journals Transforming Growth Factor β1 Ameliorates Intestinal Epithelial Barrier Disruption by Cryptosporidium parvum In Vitro in the Absence of Mucosal T Lymphocytes

2000 ◽  
Vol 68 (10) ◽  
pp. 5635-5644 ◽  
Author(s):  
James K. Roche ◽  
Clovis A. P. Martins ◽  
Rosana Cosme ◽  
Ronald Fayer ◽  
Richard L. Guerrant
Keyword(s):  
Epithelial Cell ◽  
Barrier Function ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Epithelial Barrier ◽  
Intestinal Epithelial ◽  
Epithelial Barrier Function ◽  
Epithelial Monolayers ◽  
Tgf Β1

ABSTRACT Exposure to oocysts of the protozoan Cryptosporidium parvum causes intestinal epithelial cell dysfunction in vivo and in vitro, but effective means by which mucosal injury might be prevented remain unclear. We examined the ability of transforming growth factor β1 (TGF-β1)—a cytokine synthesized and released by cells in the intestine—to preserve the barrier function of human colonic epithelia when challenged with C. parvum oocysts and then studied the mechanisms involved. Epithelial barrier function was monitored electrophysiologically, receptors for TGF-β1 were localized by confocal microscopy, and TGF-β1-induced protein kinase C activation was detected intracellularly by translocation of its α isozyme. TGF-β1 alone enhanced intestinal epithelial barrier function, while exposure to C. parvum oocysts (≥105/monolayer) markedly reduced barrier function to ≤40% of that of the control. When epithelial monolayers were pretreated with TGF-β1 at 5.0 ng/ml, the barrier-disrupting effect ofC. parvum oocysts was almost completely abrogated for 96 h. Further investigation showed that (i) the RI and RII receptors for TGF-β1 were present on 55 and 65% of human epithelial cell line cells, respectively, over a 1-log-unit range of receptor protein expression, as shown by flow cytometry and confirmed by confocal microscopy; (ii) only basolateral and not apical TGF-β1 exposure of the polarized epithelial monolayer resulted in a protective effect; and (iii) TGF-β1 had no direct effect on the organism in reducing its tissue-disruptive effects. In exploring mechanisms to account for the barrier-preserving effects of TGF-β1 on epithelium, we found that the protein kinase C pathway was activated, as shown by translocation of its 80-kDa α isozyme within 30 s of epithelial exposure to TGF-β1; the permeability of epithelial monolayers to passage of macromolecules was reduced by 42% with TGF-β1, even in the face of active protozoal infection; and epithelial cell necrosis monitored by lactate dehydrogenase release was decreased by 50% 70 h after oocyst exposure. Changes in epithelial function, initiated through an established set of surface receptors, likely accounts for the remarkable barrier-sparing effect of nanogram-per-milliliter concentrations of TGF-β1 when human colonic epithelium is exposed to an important human pathogen, C. parvum.

scholarly journals H19Long Noncoding RNA Regulates Intestinal Epithelial Barrier Function via MicroRNA 675 by Interacting with RNA-Binding Protein HuR

2016 ◽  
Vol 36 (9) ◽  
pp. 1332-1341 ◽  
Author(s):  
Tongtong Zou ◽  
Suraj K. Jaladanki ◽  
Lan Liu ◽  
Lan Xiao ◽  
Hee Kyoung Chung ◽  
...  
Keyword(s):  
Barrier Function ◽  
Noncoding Rna ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Epithelial Barrier ◽  
Intestinal Epithelial ◽  
Intestinal Epithelial Barrier ◽  
Epithelial Barrier Function ◽  
E Cadherin

The disruption of the intestinal epithelial barrier function occurs commonly in various pathologies, but the exact mechanisms responsible are unclear. TheH19long noncoding RNA (lncRNA) regulates the expression of different genes and has been implicated in human genetic disorders and cancer. Here, we report thatH19plays an important role in controlling the intestinal epithelial barrier function by serving as a precursor for microRNA 675 (miR-675).H19overexpression increased the cellular abundance of miR-675, which in turn destabilized and repressed the translation of mRNAs encoding tight junction protein ZO-1 and adherens junction E-cadherin, resulting in the dysfunction of the epithelial barrier. Increasing the level of the RNA-binding protein HuR in cells overexpressingH19prevented the stimulation of miR-675 processing fromH19, promoted ZO-1 and E-cadherin expression, and restored the epithelial barrier function to a nearly normal level. In contrast, the targeted deletion of HuR in intestinal epithelial cells enhanced miR-675 production in the mucosa and delayed the recovery of the gut barrier function after exposure to mesenteric ischemia/reperfusion. These results indicate thatH19interacts with HuR and regulates the intestinal epithelial barrier function via theH19-encoded miR-675 by altering ZO-1 and E-cadherin expression posttranscriptionally.

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