scholarly journals Fas activates the JNK pathway in human colonic epithelial cells: lack of a direct role in apoptosis

1999 ◽  
Vol 276 (3) ◽  
pp. G599-G605 ◽  
Author(s):  
Maria T. Abreu-Martin ◽  
Andrew A. Palladino ◽  
Mary Faris ◽  
Nelson M. Carramanzana ◽  
André E. Nel ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Intestinal Mucosa ◽  
Binding Activity ◽  
Kinase Assay ◽  
Complex Signal ◽  
Western Blots ◽  
Colonic Epithelial Cells ◽  
Jnk Pathway ◽  
Ifn Γ ◽  
Ht 29

Fas is expressed constitutively by colonic epithelial cells, and its ligand is expressed by intraepithelial and lamina propria lymphocytes. Fas ligation induces apoptosis in colonic epithelial cells and is implicated in the epithelial damage seen in ulcerative colitis. To understand the pleiotropic effects of Fas in the intestinal mucosa, we have examined signaling pathways activated by Fas in HT-29 colonic epithelial cells. HT-29 cells were stimulated with anti-Fas in the presence or absence of interferon-γ (IFN-γ). Activation of mitogen-activated protein kinase pathways was assessed by kinase assay, Western blots, and promoter-reporter assays. Electromobility shift assays were used to assess activator protein-1 (AP-1) binding activity. IFN-γ increases expression of Fas on HT-29 cells. Signaling via Fas receptor, as determined by induction of c-Jun NH2-terminal kinase (JNK) activity and transcriptional activation of AP-1, is enhanced in IFN-γ-primed cells. Dominant-interfering mutants of the JNK pathway do not block Fas-mediated apoptosis. Signaling through Fas results in activation of JNK and AP-1 binding activity that is increased in the presence of IFN-γ. Inhibition of JNK does not block Fas-mediated apoptosis in these cells. Fas-Fas ligand interactions in the intestinal mucosa may lead to complex signal transduction cascades and gene regulation that culminate in apoptosis, cytokine secretion, or other novel functions.

IFN-γ induces transcription but not secretion of IL-18 in human colonic epithelial cells

2001 ◽  
Vol 120 (5) ◽  
pp. A704-A704
Author(s):  
S KIESSLING ◽  
K SCHIOTTMANN ◽  
W FALK ◽  
T ANDUS ◽  
J SCHOELMERICH ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Colonic Epithelial Cells ◽  
Ifn Γ

IL-8 secretion and neutrophil activation by HT-29 colonic epithelial cells

1994 ◽  
Vol 267 (6) ◽  
pp. G991-G997 ◽  
Author(s):  
C. P. Kelly ◽  
S. Keates ◽  
D. Siegenberg ◽  
J. K. Linevsky ◽  
C. Pothoulakis ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Intestinal Inflammation ◽  
Neutrophil Migration ◽  
Intercellular Adhesion Molecule ◽  
Northern Hybridization ◽  
Intercellular Adhesion Molecule 1 ◽  
Colonic Epithelial Cells ◽  
Neutrophil Extravasation ◽  
Cytokine Stimulation ◽  
Ht 29

This study examines the ability of HT-29 human colonic epithelial cells to stimulate neutrophil migration and adhesion. Interleukin-8 (IL-8), a potent neutrophil chemoattractant, was detected in conditioned media from both unstimulated (1.1 ng/ml) and IL-1 beta-stimulated (16.1 ng/ml) HT-29 cultures. Conditioned medium from IL-1 beta-exposed HT-29 cells stimulated neutrophil migration (395% of control, P < 0.01), and this effect was completely inhibited by anti-IL-8 antibody. HT-29 medium also induced shedding of neutrophil L-selectin and increased expression of neutrophil CD11/CD18 adhesion receptors. Coculture of HT-29 cells with human endothelial cell monolayers resulted in increased neutrophil transendothelial migration (169% of control, P < 0.01), which was blocked by both anti-IL-8 and anti-CD18 antibody. Northern hybridization analysis demonstrated increased levels of mRNA for IL-8 and intercellular adhesion molecule-1 (ICAM-1) in cytokine-treated HT-29 cells. Cytokine stimulation of HT-29 monolayers was also associated with increased neutrophil adhesion to these cells. Neutrophil-HT-29 cell adhesion was blocked by monoclonal antibodies to neutrophil CD18 or to ICAM-1 on the HT-29 cells (86% and 56% inhibition, respectively, P < 0.01 for both). These data suggest that IL-8 secretion by activated colonic epithelial cells may contribute to neutrophil extravasation and tissue infiltration in intestinal inflammation.

scholarly journals Clostridium butyricum Induces the Production and Glycosylation of Mucins in HT-29 Cells

2021 ◽  
Vol 11 ◽  
Author(s):  
Qi Lili ◽  
Lu Xiaohui ◽  
Mao Haiguang ◽  
Wang Jinbo
Keyword(s):  
Rna Interference ◽  
Epithelial Cells ◽  
Commensal Bacteria ◽  
Human Intestine ◽  
Commensal Bacterium ◽  
Colonic Epithelial Cells ◽  
Mechanism Of Interaction ◽  
Positive Functions ◽  
First Time ◽  
Ht 29

C. butyricum is a common gut commensal bacterium, which has many positive functions in human intestine. In this study, we investigated the effects of monosaccharide and its derivatives on the adhesion of C. butyricum to the mucus of HT-29 cells. RNA interference was performed to assess the roles of MUC2 and glycan in the adhesion of C. butyricum to HT-29 cells. The effects of C. butyricum on the glycosylation of mucins were assayed with fluorescence microscope. The expression levels of mucins and glycotransferases were also determined. The results showed that C. butyricum could adhere to the mucins secreted by HT-29 cells. Several kinds of monosaccharides inhibited the adhesion of C. butyricum to HT-29 cells, which suggested that the mucus glycan was the attaching sites of this bacterium. Knockdown of MUC2, FUT2 or GALNT7 significantly decreased the numbers of the bacteria adhering to HT-29 cells. When colonizing on the surface of HT-29 cells, C. butyricum could increase the production of mucins, promote the expression of glycotransferase, and induce the glycosylation of mucins. These results demonstrated that the glycan of mucus played important roles in the adhesion of C. butyricum to HT-29 cells. This study indicates for the first time that C. butyricum possesses the ability to modulate the glycosylation profile of mucus secreted by HT-29 cells. These findings contribute to understanding the mechanism of interaction between colonic epithelial cells and commensal bacteria.

Cycle of VIP in the human transformed colonic epithelial cells (HT-29) in culture

1985 ◽  
Vol 10 ◽  
pp. S33 ◽  
Author(s):  
J.C. Marie ◽  
D. Hui Bon Hoa ◽  
G. Hejblum ◽  
G. Rosselin
Keyword(s):  
Epithelial Cells ◽  
Colonic Epithelial Cells ◽  
Ht 29

Neurotensin stimulates IL-8 expression in human colonic epithelial cells through Rho GTPase-mediated NF-κB pathways

2003 ◽  
Vol 284 (6) ◽  
pp. C1397-C1404 ◽  
Author(s):  
Dezheng Zhao ◽  
Sabina Kuhnt-Moore ◽  
Huiyan Zeng ◽  
Jack S. Wu ◽  
Mary P. Moyer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Production ◽  
Rho Gtpase ◽  
Intestinal Inflammation ◽  
Binding Activity ◽  
Dominant Negative ◽  
Colonic Epithelial Cells ◽  
Mapk Phosphorylation ◽  
Dna Binding Activity ◽  
Rho Family

Neurotensin (NT), a neuropeptide highly expressed in the gastrointestinal tract, participates in the pathophysiology of intestinal inflammation. We recently showed that NT stimulates interleukin-8 (IL-8) expression in NCM460 nontransformed human colonic epithelial cells via both mitogen-activating protein kinase (MAPK)- and NF-κB-dependent pathways. However, the molecular mechanism by which NT induces expression of proinflammatory cytokines such as IL-8 has not been investigated. In this study we show that inhibition of endogenous Rho family proteins (RhoA, Rac1, and Cdc42) by their respective dominant negative mutants inhibits NT-induced IL-8 protein production and promoter activity. Western blot experiments demonstrated that NT strongly activated RhoA, Rac1, and Cdc42. Overexpression of the dominant negative mutants of RhoA, Rac1, and Cdc42 significantly inhibited NT-induced NF-κB-dependent reporter gene expression and NF-κB DNA binding activity. NT also stimulated p38 MAPK phosphorylation, and overexpression of dominant negative mutants of RhoA, Rac1, and Cdc42 did not significantly alter p38 and ERK1/2 phosphorylation in response to NT. Together, our findings indicate that NT-stimulated IL-8 expression is mediated via a Rho-dependent NF-κB-mediated pathway.

Cycle of VIP in the human transformed colonic epithelial cells (HT-29) in culture

Peptides ◽  
1986 ◽  
Vol 7 ◽  
pp. 129-135 ◽  
Author(s):  
J.C. Marie ◽  
D. Hui Bon Hoa ◽  
G. Hejblum ◽  
G. Rosselin
Keyword(s):  
Epithelial Cells ◽  
Colonic Epithelial Cells ◽  
Ht 29

Human intestinal intraepithelial lymphocytes and epithelial cells coinduce interleukin-8 production through the CD2-CD58 interaction

2009 ◽  
Vol 296 (3) ◽  
pp. G671-G677 ◽  
Author(s):  
Ellen C. Ebert ◽  
Asit Panja ◽  
Rajalakshmi Praveen
Keyword(s):  
Epithelial Cells ◽  
Cell Types ◽  
Interferon Γ ◽  
Intraepithelial Lymphocytes ◽  
Cell Contact ◽  
Tnf Α ◽  
Active Transcription ◽  
Ifn Γ ◽  
Factor Α ◽  
Ht 29

Human intestinal CD3+TCRαβ+CD8+ intraepithelial lymphocytes (IELs) are intimately associated with epithelial cells (ECs) through binding of CD103 to E-cadherin. How these two cell types functionally interact is largely unknown. IEL-EC cross talk was determined using HT-29 cells as the model EC and IL-8 as the readout. IL-8 was derived from both cell types and synergistically increased when the cells were combined. This synergistic effect required active transcription by both IELs and HT-29 cells. Cell contact was required as shown by the loss of the synergistic increase in IL-8 when the two cell types were separated by Transwells. Specifically, IL-8 release required the binding of CD2 on the IELs to CD58 on the HT-29 cells. The association of the CD3/TCR complex with major histocompatibility antigen class I antigens was not involved. Antibody neutralization of tumor necrosis factor-α (TNF-α), but not interferon-γ (IFN-γ), resulted in increased IL-8 production by the coculture. Although both TNF-α and IFN-γ increased IL-8 synthesis and CD58 expression by the HT-29 cells, only IFN-γ reduced IL-8 production by IELs. IL-8 production by either cell type involved phosphorylation of p38 and JNK. In summary, the synergistic synthesis of IL-8 occurs when IELs are stimulated through the CD2 pathway by CD58 on HT-29 cells, resulting in TNF-α release that, in turn, augments IL-8 synthesis and CD58 expression by the HT-29 cells.

Differential effects of deoxycholic acid and taurodeoxycholic acid on NF-κB signal transduction and IL-8 gene expression in colonic epithelial cells

2004 ◽  
Vol 286 (6) ◽  
pp. G1000-G1008 ◽  
Author(s):  
M. Mühlbauer ◽  
B. Allard ◽  
A. K. Bosserhoff ◽  
S. Kiessling ◽  
H. Herfarth ◽  
...  
Keyword(s):  
Gene Expression ◽  
Signal Transduction ◽  
Epithelial Cells ◽  
Molecular Mechanisms ◽  
Deoxycholic Acid ◽  
Nuclear Translocation ◽  
Binding Activity ◽  
Dominant Negative ◽  
Dependent Manner ◽  
Colonic Epithelial Cells

Several effects of bile acids (BAs) on colonic epithelial cells (CECs) have been described, including induction of proliferation and apoptosis. Some of these effects are mediated through activation of the NF-κB transcriptional system. In this study, we investigated the molecular mechanisms underlying the BA-induced gene expression in CECs. The human CEC line HT-29 and primary human CECs were treated with dilutions of salts of deoxycholic acid (DCA) and taurodeoxycholic acid (TDCA). NF-κB binding activity was analyzed with EMSA, RelA translocation with immunofluorescence, and IκBα- and RelA-phosphorylation with Western blot analysis. IL-8 mRNA and protein expression were assessed by quantitative PCR and ELISA. Functional impact of NF-κB activation was determined by blocking the proteasome activity with MG132 or by preventing IKK activity with a dominant-negative IKKβ delivered by adenoviral dominant-negative (dn) IKKβ (Ad5dnIKKβ). DCA and TDCA induced IL-8 expression in a dose- and time-dependent manner. It is interesting that DCA but not TDCA induced IκBα-phophorylation, RelA translocation, and NF-κB binding activity. Accordingly, the proteasome inhibitor MG132 blocked DCA- but not TDCA-induced IL-8 gene expression. In contrast, TDCA-induced IL-8 gene expression correlated with enhanced RelA phosphorylation, which was blocked by Ad5dnIKKβ. Our data suggest that DCA-induced signal transduction mainly utilized the IκB degradation and RelA nuclear translocation pathway, whereas TDCA primarily induced IL-8 gene expression through RelA phosphorylation. These differences may have implications for the understanding of the pathophysiology of inflammation and carcinogenesis in the gut.

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