Regulation of epithelial syndecan 1 expression by inflammatory cytokines
Download
1 / 27

Regulation of epithelial syndecan-1 expression by inflammatory cytokines - PowerPoint PPT Presentation


  • 499 Views
  • Uploaded on

Regulation of epithelial syndecan-1 expression by inflammatory cytokines. Richard M. Day, Tracey J. Mitchell, Stella C. Knight, Alastair Forbes Cytokine 21 (2003) 224–233. Introduction. Inflammatory bowel disease (IBD).

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'Regulation of epithelial syndecan-1 expression by inflammatory cytokines' - arleen


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
Regulation of epithelial syndecan 1 expression by inflammatory cytokines l.jpg

Regulation of epithelial syndecan-1 expression by inflammatory cytokines

Richard M. Day, Tracey J. Mitchell, Stella C. Knight, Alastair Forbes

Cytokine 21 (2003) 224–233


Introduction l.jpg
Introduction inflammatory cytokines


Inflammatory bowel disease ibd l.jpg
Inflammatory bowel disease (IBD) inflammatory cytokines

Inflammatory bowel disease (IBD) refers to the condition that results when cells involved in inflammation and immune response are called into the lining of the GI tract.

IBD is characterized by chronic intestinal inflammation that results in clinical symptoms such as diarrhea, bleeding, abdominal pain, fever, joint pain, and weight loss.

Dysregulation of epithelial glycosaminoglycans (GAGs) occurs in IBD

Syndecans are a class of heparan sulphate proteoglycans that mediate both cell adhesion and growth factor binding via GAG side chains.


Syndecan 1 l.jpg
Syndecan-1 inflammatory cytokines

Syndecan-1(CD138)

The best characterized of the four syndeacan core proteins binding to variety of components of extracellular matrix, including collagen type1 ,3,5 and fibronectin.

Syndecan-1 may regulate ligand-dependent activation of cell surface growth factor at two dimernsional surface of plasma membrane

Fibroblast growth factor(bFGF), hepatocyte growth factor,platelet-derived growth factor,vascular endothelial growth factor,and etc.


Slide6 l.jpg

Mucosal cytokine may regulate epithelial cell function. inflammatory cytokines

Reduced expression of syndecan-1 observed in IBD results from the increased presence of inflammatory cytokines.


Slide7 l.jpg

  • Cell culture inflammatory cytokines

    • Human colorectal carcinoma epithelial cell lines HT29/219 and T84

  • Cytotoxicity assay

    • The cytokine effects of TNF-a, IL-1b and IL-6.

  • Flow cytometry analysis

    • Epithelial cell surface syndecan-1 expression analysis

  • Co-culture with peripheral blood mononuclear cells

    • Co-culture of PBMC and epithelial (HT29) cell were stimulated with 20 ng/ml PMA and 2mM ionomycin for 24hr.

    • PMA phorbol 12-mryristate 13-acetate


  • Slide8 l.jpg

    Detection of soluble syndecan-1 inflammatory cytokinesBy ELISA

    Immunocytochemistry

    RNA extraction and amplification by reverse transcription-polymerase chain reaction

    Statistical analysis


    Results l.jpg
    Results inflammatory cytokines


    Slide10 l.jpg

    Fig. 1 inflammatory cytokines

    unstimulated

    5 ng/ml TNF-a

    75 ng/ml TNF-a

    IgG1 isotype control


    Slide11 l.jpg

    IL-6 inflammatory cytokines

    TNF-a

    IL-1b

    TNF-a

    B,C,D HT29 cell

    E T84 cell


    Slide12 l.jpg

    Cell variability --- ApoAlert cytotoxicity assay inflammatory cytokines

    Above 86% for all concentrations of cytokine studied.


    Slide13 l.jpg

    Fig. 2A inflammatory cytokines

    HT29 cell treated with 5 ng/ml TNF-a for the various time


    Slide14 l.jpg

    Fig. 2B inflammatory cytokines


    Slide15 l.jpg

    Fig. 3 inflammatory cytokines

    HT29 cell treated with TNF-a, IL-1b and IL-6 for 24h

    Syndecan-1 expression reduced to 49.7%

    Syndecan-1 expression reduced to 64.3%


    Slide16 l.jpg

    Fig. 4 inflammatory cytokines

    Increase 69.3%

    Cytokine stimulation induced syndecan-1 shedding from the cell surface

    Increase 8.1%

    Increase 17.1%


    Slide17 l.jpg

    Fig. 5 inflammatory cytokines


    Slide18 l.jpg

    Fig. 6 inflammatory cytokines

    Co-culture HT29 cell and PBMC

    PBMC endogenous inflammatory cytokine

    PMA increase syndecan-1 suppression


    Discussion l.jpg
    Discussion inflammatory cytokines


    Slide20 l.jpg

    Wound repair

    other model without inflammation

    (up-regulation)

    The expression of

    syndecan-1

    Increased syndecan-1 expression during wound repair is thought to facilitate growth factor binding and wound healing.

    The reduced expression in IBD maybe related to the presence of inflammatory cytokines in mucosa.


    Slide21 l.jpg

    Does-dependent down-regulate syndecan-1 mRNA mucosa of the patients with IBD

    Stable expression of syndecan-1

    TNF-a, IL-1b and IL6 stimulate

    Reduced Syndecan-1 expression

    HT29 cell

    T84 cell

    IL-6 to cause a reduction of syndecan-1 expression in murine B-lymphoid cells  cell-type specific


    Slide22 l.jpg

    Epithelial cell adhesion molecules mucosa of the patients with IBD

    Syndecan-1, E-cadherin

    Inflammatory cytokine ex. TNF-a IL-1

    dysregulation

    Expression of E-cadherin is dependent on syndecan-1 expression and vice versa.

    Cytokine induce loss of cell-adhesion through reduced syndecan-1 and/or E-caderine destablizes the epithelial barrier.

    Increase intestinal permeability in IBD


    Slide23 l.jpg

    Shedding of syndeacan-1 ectodomain into the culture medium of HT29 cell monolayers was significantly increase following the addition of TNF-a .

    The increase syndecan-1, shedding following stimulation correlated with the loss of membrane syndecan-1 expression

    Some syndecan-1 detected may also have been released as the intact proteoglycan from dead cell


    Slide24 l.jpg

    Release inflammatory cytokine of HT29 cell monolayers was significantly increase following the addition of TNF-a .

    block

    PBMC

    Reduced syndecan-1 expression

    Co-culture

    HT29 cell

    Increased ICAM-1 expression

    HT29 cell

    + TNF-a and IL-1b

    The loss of GAG expression in p’t with IBD is associated with increased density of TNF-a


    Slide25 l.jpg

    This study demonstrates dysregulation of epithelial syndecan-1 expression by inflammatory cytokines and may have implications for mucosal ulcer healing.


    Slide26 l.jpg

    Inflammatory bowel disease refers to the condition that results when cells involved in inflammation and immune response are called into the lining of the GI tract. This infiltration thickens the bowel lining and interferes with absorption and motility (the ability of the bowel to contract and move food). With abnormal ability to contract and abnormal ability to absorb, the bowel’s function is disrupted. Chronic vomiting results if the infiltration is in the stomach or higher areas of the small intestine. A watery diarrhea with weight loss results if the infiltration is in the lower small intestine. A mucous diarrhea with fresh blood (colitis)  results if the infiltration occurs in the large intestine. Of course, the entire tract from top to bottom may be involved. Many people confuse Inflammatory Bowel Disease with “Irritable Bowel Syndrome,” a stress-related diarrhea problem. Treatment for “IBS” is aimed at stress; it is a completely different condition from “IBD.”


    Slide27 l.jpg

    These symptoms can range from mild to severe, and may gradually and subtly develop from an initial minor discomfort, or may present themselves suddenly with acute intensity.


    ad