scholarly journals Angulin-1 (LSR) Affects Paracellular Water Transport, However Only in Tight Epithelial Cells

2021 ◽  
Vol 22 (15) ◽  
pp. 7827
Author(s):  
Carlos Ayala-Torres ◽  
Susanne M. Krug ◽  
Rita Rosenthal ◽  
Michael Fromm
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Water Transport ◽  
Cell Lines ◽  
Water Permeability ◽  
Water Flux ◽  
Epithelial Cell Line ◽  
Osmotic Gradient ◽  
The Impact ◽  
Ht 29

Water transport in epithelia occurs transcellularly (aquaporins) and paracellularly (claudin-2, claudin-15). Recently, we showed that downregulated tricellulin, a protein of the tricellular tight junction (tTJ, the site where three epithelial cells meet), increased transepithelial water flux. We now check the hypothesis that another tTJ-associated protein, angulin-1 (alias lipolysis-stimulated lipoprotein receptor, LSR) is a direct negative actuator of tTJ water permeability depending on the tightness of the epithelium. For this, a tight and an intermediate-tight epithelial cell line, MDCK C7 and HT-29/B6, were stably transfected with CRISPR/Cas9 and single-guide RNA targeting angulin-1 and morphologically and functionally characterized. Water flux induced by an osmotic gradient using 4-kDa dextran caused water flux to increase in angulin-1 KO clones in MDCK C7 cells, but not in HT-29/B6 cells. In addition, we found that water permeability in HT-29/B6 cells was not modified after either angulin-1 knockout or tricellulin knockdown, which may be related to the presence of other pathways, which reduce the impact of the tTJ pathway. In conclusion, modulation of the tTJ by knockout or knockdown of tTJ proteins affects ion and macromolecule permeability in tight and intermediate-tight epithelial cell lines, while the transepithelial water permeability was affected only in tight cell lines.

scholarly journals Tricellulin Effect on Paracellular Water Transport

2019 ◽  
Vol 20 (22) ◽  
pp. 5700 ◽  
Author(s):  
Carlos Ayala-Torres ◽  
Susanne M. Krug ◽  
Jörg D. Schulzke ◽  
Rita Rosenthal ◽  
Michael Fromm
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Water Transport ◽  
Water Permeability ◽  
Water Flux ◽  
Epithelial Cell Line ◽  
Osmotic Gradient ◽  
Direct Role ◽  
Main Protein ◽  
Molecular Components

In epithelia, large amounts of water pass by transcellular and paracellular pathways, driven by the osmotic gradient built up by the movement of solutes. The transcellular pathway has been molecularly characterized by the discovery of aquaporin membrane channels. Unlike this, the existence of a paracellular pathway for water through the tight junctions (TJ) was discussed controversially for many years until two molecular components of paracellular water transport, claudin-2 and claudin-15, were identified. A main protein of the tricellular TJ (tTJ), tricellulin, was shown to be downregulated in ulcerative colitis leading to increased permeability to macromolecules. Whether or not tricellulin also regulates water transport is unknown yet. To this end, an epithelial cell line featuring properties of a tight epithelium, Madin-Darby canine kidney cells clone 7 (MDCK C7), was stably transfected with small hairpin RNA (shRNA) targeting tricellulin, a protein of the tTJ essential for the barrier against passage of solutes up to 10 kDa. Water flux was induced by osmotic gradients using mannitol or 4 and 40 kDa-dextran. Water flux in tricellulin knockdown (KD) cells was higher compared to that of vector controls, indicating a direct role of tricellulin in regulating water permeability in a tight epithelial cell line. We conclude that tricellulin increases water permeability at reduced expression.

scholarly journals A Comparative Study on the Model of PM2.5 Direct or Indirect Interaction with Bronchial Epithelial Cells.

2021 ◽  
Author(s):  
Yan Wang ◽  
Xin Zuo ◽  
Fuyang Jiang ◽  
Lin Hou ◽  
Qiyue Jiang ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Damage ◽  
Direct Interaction ◽  
Bronchial Epithelial Cell ◽  
Human Bronchial Epithelial Cell ◽  
Exposure Model ◽  
Epithelial Cell Line ◽  
Bronchial Epithelial ◽  
The Impact

Abstract The impact of PM2.5 on epithelial cells is a pivotal process leading to many lung pathological changes and pulmonary diseases. In addition to PM2.5 direct interaction with epithelia, macrophages that engulf PM2.5 may also influence the function of epithelial cells. However, among the toxic researches of PM2.5, there is a lack of evaluation of direct or indirect exposure model on human bronchial epithelial cell against PM2.5. In this present research, PM2.5-exposed human bronchial epithelial cell line (BEAS-2B) serves as the direct interaction model, while the contrast is to indirect stimulation model, which takes advantage of transwell co-culture system to carry out that PM2.5 is promptly contacted with macrophages rather than BEAS-2B. By comparing these two modes of interaction, we determined the viability of BEAS-2B and mRNA and/or protein expression profile of transcription factors Nrf2,NF-kB and according inflammatory indicators, with a view to evaluating the effects of different interaction modes of PM2.5 on epithelial cell damage in vitro. We have found that macrophage involvement may protect epithelia from PM2.5 cytotoxic effect, while strengthen the inflammation response.

scholarly journals CD14- and Toll-Like Receptor-Dependent Activation of Bladder Epithelial Cells by Lipopolysaccharide and Type 1 Piliated Escherichia coli

2003 ◽  
Vol 71 (3) ◽  
pp. 1470-1480 ◽  
Author(s):  
Joel D. Schilling ◽  
Steven M. Martin ◽  
David A. Hunstad ◽  
Kunal P. Patel ◽  
Matthew A. Mulvey ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Lines ◽  
Epithelial Cell Line ◽  
Renal Epithelial Cell ◽  
E Coli ◽  
Epithelial Cell Lines ◽  
Renal Epithelial Cell Line

ABSTRACT The gram-negative bacterium Escherichia coli is the leading cause of urinary tract infection. The interaction between type 1 piliated E. coli and bladder epithelial cells leads to the rapid production of inflammatory mediators, such as interleukin-6 (IL-6) and IL-8. Conflicting reports have been published in the literature regarding the mechanism by which uroepithelial cells are activated by type 1 piliated E. coli. In particular, the role of lipopolysaccharide (LPS) in these responses has been an area of significant debate. Much of the data arguing against LPS-mediated activation of bladder epithelial cells have come from studies using a renal epithelial cell line as an in vitro model of the urinary epithelium. In this report, we analyzed three bladder epithelial cell lines and demonstrated that they all respond to LPS. Furthermore, the LPS responsivity of the cell lines directly correlated with their ability to generate IL-6 after E. coli stimulation. The LPS receptor complex utilized by the bladder epithelial cell lines included CD14 and Toll-like receptors, and signaling involved the activation of NF-κB and p38 mitogen-activated protein kinase. Also, reverse transcription-PCR analysis demonstrated that bladder epithelial cells express CD14 mRNA. Thus, the molecular machinery utilized by bladder epithelial cells for the recognition of E. coli is very similar to that described for traditional innate immune cells, such as macrophages. In contrast, the A498 renal epithelial cell line did not express CD14, was hyporesponsive to LPS stimulation, and demonstrated poor IL-6 responses to E. coli.

scholarly journals The Pseudomonas aeruginosa Exopolysaccharide Psl Facilitates Surface Adherence and NF-κB Activation in A549 Cells

mBio ◽  
2010 ◽  
Vol 1 (3) ◽  
Author(s):  
Matthew S. Byrd ◽  
Bing Pang ◽  
Meenu Mishra ◽  
W. Edward Swords ◽  
Daniel J. Wozniak
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
A549 Cells ◽  
Luciferase Reporter ◽  
Epithelial Cell Line ◽  
Bacterial Cells ◽  
Burn Wound ◽  
The Impact

ABSTRACTIn order for the opportunistic Gram-negative pathogenPseudomonas aeruginosato cause an airway infection, the pathogen interacts with epithelial cells and the overlying mucous layer. We examined the contribution of the biofilm polysaccharide Psl to epithelial cell adherence and the impact of Psl on proinflammatory signaling by flagellin. Psl has been implicated in the initial attachment ofP. aeruginosato biotic and abiotic surfaces, but its direct role in pathogenesis has not been evaluated (L. Ma, K. D. Jackson, R. M. Landry, M. R. Parsek, and D. J. Wozniak, J. Bacteriol. 188:8213–8221, 2006). Using an NF-κB luciferase reporter system in the human epithelial cell line A549, we show that both Psl and flagellin are necessary for full activation of NF-κB and production of the interleukin 8 (IL-8) chemokine. We demonstrate that Psl does not directly stimulate NF-κB activity, but indirectly as a result of increasing contact between bacterial cells and epithelial cells, it facilitates flagellin-mediated proinflammatory signaling. We confirm differential adherence of Psl and/or flagellin mutants by scanning electron microscopy and identify Psl-dependent membrane structures that may participate in adherence. Although we hypothesized that Psl would protectP. aeruginosafrom recognition by the epithelial cell line A549, we instead observed a positive role for Psl in flagellin-mediated NF-κB activation, likely as a result of increasing contact between bacterial cells and epithelial cells.IMPORTANCEPseudomonas aeruginosais the predominant airway pathogen causing morbidity and mortality in individuals affected by the genetic disease cystic fibrosis.P. aeruginosacan also cause severe pneumonia, burn wound infections, and sepsis, making its overall impact on human health significant. The attachment ofP. aeruginosato host tissues, often leading to recalcitrant biofilm infections, and inflammation induced by flagellin are both important mechanisms of virulence. We explored the role of the biofilm polysaccharide Psl in the pathogenesis ofP. aeruginosaand found that Psl is required for surface adherence to A549 epithelial cells, and as an adhesin, it facilitates flagellin-mediated NF-κB activation. This work was done to better understand the initial events of infection and revealed that a biofilm polysaccharide contributes to inflammation in a novel manner.

scholarly journals Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.

1988 ◽  
Vol 106 (3) ◽  
pp. 761-771 ◽  
Author(s):  
P Boukamp ◽  
R T Petrussevska ◽  
D Breitkreutz ◽  
J Hornung ◽  
A Markham ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Human Skin ◽  
Cell Lines ◽  
Hacat Cell ◽  
Epithelial Cell Line ◽  
Spontaneous Transformation ◽  
Chromosomal Alterations ◽  
Hacat Cell Line

In contrast to mouse epidermal cells, human skin keratinocytes are rather resistant to transformation in vitro. Immortalization has been achieved by SV40 but has resulted in cell lines with altered differentiation. We have established a spontaneously transformed human epithelial cell line from adult skin, which maintains full epidermal differentiation capacity. This HaCaT cell line is obviously immortal (greater than 140 passages), has a transformed phenotype in vitro (clonogenic on plastic and in agar) but remains nontumorigenic. Despite the altered and unlimited growth potential, HaCaT cells, similar to normal keratinocytes, reform an orderly structured and differentiated epidermal tissue when transplanted onto nude mice. Differentiation-specific keratins (Nos. 1 and 10) and other markers (involucrin and filaggrin) are expressed and regularly located. Thus, HaCaT is the first permanent epithelial cell line from adult human skin that exhibits normal differentiation and provides a promising tool for studying regulation of keratinization in human cells. On karyotyping this line is aneuploid (initially hypodiploid) with unique stable marker chromosomes indicating monoclonal origin. The identity of the HaCaT line with the tissue of origin was proven by DNA fingerprinting using hypervariable minisatellite probes. This is the first demonstration that the DNA fingerprint pattern is unaffected by long-term cultivation, transformation, and multiple chromosomal alterations, thereby offering a unique possibility for unequivocal identification of human cell lines. The characteristics of the HaCaT cell line clearly document that spontaneous transformation of human adult keratinocytes can occur in vitro and is associated with sequential chromosomal alterations, though not obligatorily linked to major defects in differentiation.

Studying Permeability in a Commonly Used Epithelial Cell Line: T84 Intestinal Epithelial Cells

2011 ◽  
pp. 115-137 ◽  
Author(s):  
Rino P. Donato ◽  
Adaweyah El-Merhibi ◽  
Batjargal Gundsambuu ◽  
Kai Yan Mak ◽  
Emma R. Formosa ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Intestinal Epithelial Cells ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial

The lack of attachment of transformed embryonic lung epithelial cells to collagen I is corrected by fibronectin and FXIII

1986 ◽  
Vol 86 (1) ◽  
pp. 95-107
Author(s):  
M. Paye ◽  
C.M. Lapiere
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Lung Epithelial Cells ◽  
Collagen I ◽  
Epithelial Cell Line ◽  
Embryonic Lung ◽  
Lung Epithelial ◽  
Minimal Concentration ◽  
Pulmonary Epithelial Cell

PER cells, a transformed pulmonary epithelial cell line that adhered to a large extent to a fibronectin substratum, were found to be attachment-deficient to collagen I. Although fibronectin can bind to collagen I monomers and polymers, the addition of exogenous fibronectin in the attachment medium induced the adhesion of these cells to collagen I polymers but not to monomers. By adding the transglutaminase of blood coagulation, FXIII, in the presence of fibronectin, the attachment of PER cells to collagen I monomers could be recovered while the minimal concentration of fibronectin needed to promote their adhesion to polymers was lowered. These studies indicate that FXIII enhances the fibronectin-mediated attachment of PER cells to collagen I.

Basolateral secretion of kappa light chain in the polarised epithelial cell line, Caco-2

1989 ◽  
Vol 94 (2) ◽  
pp. 327-332
Author(s):  
E.J. Hughson ◽  
D.F. Cutler ◽  
C.R. Hopkins
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Light Chain ◽  
Intracellular Trafficking ◽  
Epithelial Cell Line ◽  
Kappa Light Chain ◽  
Immunoglobulin Kappa ◽  
Selective Delivery ◽  
Secretory Products

The immunoglobulin kappa light chain is constitutively secreted in non-polarised cells. It is therefore unlikely to display any of the signals thought to be required for the selective delivery of proteins to the apical or basolateral borders of polarised epithelial cells. We have transfected the gene for the kappa light chain into a polarised epithelial cell line (Caco-2) and shown that it is secreted predominantly from the basolateral surface. Metabolically labelled endogenous secretory products show the same polarity and we conclude, therefore, that in Caco-2 cells there is a major intracellular trafficking route to the basolateral border that requires no sorting signal.

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