liu.seSearch for publications in DiVA
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Interferon-γ enhancement of E-selectin expression on endothelial cells is inhibited by monensin
Linköping University, Department of Biomedicine and Surgery, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Biomedicine and Surgery, Clinical Chemistry. Linköping University, Faculty of Health Sciences.
1997 (English)In: Scandinavian Journal of Immunology, ISSN 0300-9475, E-ISSN 1365-3083, Vol. 46, no 4, 338-343 p.Article in journal (Refereed) Published
Abstract [en]

The expression of E-selectin reaches a maximum 4–6 h after stimulation of human umbilical vein endothelial cells (HUVEC) in vitro with tumour necrosis factor-α (TNF-α) and then declines to basal level within 24 h. If interferon-γ (IFN-γ) is added to the cell culture medium together with TNF-α the surface expression of E-selectin is augmented and prolonged in a synergistic way. The aim of the present study was to investigate if altered protein glycosylation could explain the IFN-γ induced persistent surface expression of E-selectin. SDS–PAGE analysis of HUVEC glycoproteins, metabolically radiolabelled in the carbohydrate portion, indicated that addition of IFN-γ produced an altered protein glycosylation. Lectin blot analysis using the Sambucus nigra agglutinin lectin also indicated differences in protein glycosylation when HUVEC were incubated with IFN-γ/TNF-α compared to TNF-α alone. The kinetics of surface expression of E-selectin were measured using a cell ELISA assay. When HUVEC were incubated with monensin, a potent inhibitor of late Golgi function, together with both TNF-α and IFN-γ, the additive effect of IFN-γ on E-selectin expression was almost abolished. Since monensin is known to affect glycosylation processing, this experiment suggested that the IFN-γ induced change in protein glycosylation might induce the prolonged surface expression of E-selectin. However, when HUVEC were cultured with IFN-γ/TNF-α in the presence of several different inhibitors of N-glycosylation processing, no significant effect of E-selectin expression was observed. Regulation of adhesion molecule expression after activation of endothelial cells is likely to play a pivotal role for the inflammatory response. Further studies are needed to understand the mechanisms underlying this regulation.

Place, publisher, year, edition, pages
1997. Vol. 46, no 4, 338-343 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-84629DOI: 10.1046/j.1365-3083.1997.d01-135.xOAI: oai:DiVA.org:liu-84629DiVA: diva2:560851
Available from: 2012-10-16 Created: 2012-10-16 Last updated: 2017-12-07Bibliographically approved
In thesis
1. Innate immune response in human endothelial cells: characterization and regulation of E-selectin, ICAM-I and cytokine expression and the role of Staphylococcus aureus
Open this publication in new window or tab >>Innate immune response in human endothelial cells: characterization and regulation of E-selectin, ICAM-I and cytokine expression and the role of Staphylococcus aureus
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Endothelial cells located on the vessel walls play an important role in inflammation. In response to cytokines or other inflammatory mediators such as microorganisms, endothelial cells express cell adhesion molecules such as E-selectin and ICAM-1 that are involved in binding circulating leukocytes to the endothelium. Since E-selectin is highly glycosylated one aim of this thesis was to investigate the role of N-glycosylation in E-selectin expression and function. Modification of glycosylation was obtained by culturing HUVEC in the presence of soluble glycosylation inhibitors. The result showed that E-selectin is highly glycosylated with N-Iinked, complex type oligosaccharides which were found to be important for the level of cell surface expression and turnover time. However, defect glycosylation did not affect its binding properties as revealed in a static cell-binding assay. Combinations of cytokines can produce a different expression of E-selectin. When TNF-α and IFN-γ were used to stimulate HUVEC an enhanced and prolonged expression of E-selectin was obtained compared to when HUVEC were stimulated with TNF-α alone. This effect could be blocked by monensin, which is an inhibitor of trans-Golgi network processing. This again indicated a role for posttranslational modification in regulation of E-selectin expression.

Eighteen clinical isolates of Staphylococcus aureus were analysed for their ability to induce expression of E-selectin and ICAM-1 in HUVEC. Upregulation of adhesion molecules varied between isolates; 17 isolates induced expression of E-selectin and 13 of ICAM-1. Some isolates induced a significant expression of E-selectin without stimulation of ICAM-1, whereas the opposite was not found. Bacterial viability was required for induction of the adhesion molecules. There was no correlation between ability of S. aureus to induce expression of cell adhesion molecules and methicillin susceptibility, pulse field electrophoresis patterns, biochemical characteristics, phage typing or toxin production. Nine cytokines, IL-1ß, TNF-α, IL-6, IL-8, IL-10, IL-12p70, GRO-α, GM-CSF and RANTES, all of importance in the inflammatory process, were analysed in supernatants from HUVEC stimulated with eighteen isolates of S. aureus. All isolates induced IL-6, IL-8, GRO-α, GM-CSF and RANTES. Isolates of S. aureus inducing high expression of one of these cytokines also showed high expression of the other four, indicating a possible common mechanism for regulation of the expression of these cytokines. The ability of individual S. aureus isolates to induce expression of cytokines correlated with their ability to induce expression of ICAM-1 but not E-selectin in HUVEC. No correlation between cytokine profile and S. aureus production of enterotoxin A-D, TSST-1, cytotoxicity, PFGE- and phage pattern or susceptibility to methicillin was observed. Furthermore, culture filtrate from S. aureus induced expression of ICAM-1 and IL-8 in HUVEC. The component(s) from culture filtrates were heat-stable, had a molecular weight of about 100 kDa and the effect was independent of the presence of fetal calf serum in media.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2003. 52 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 804
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-26669 (URN)11235 (Local ID)91-7373-493-4 (ISBN)11235 (Archive number)11235 (OAI)
Public defence
2003-09-26, Elsa Brändströmsalen, Hälsouniversitet, Linköping, 13:00 (Swedish)
Opponent
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2012-10-16Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full text

Authority records BETA

Strindhall, JanLundblad, ArnePåhlsson, Peter

Search in DiVA

By author/editor
Strindhall, JanLundblad, ArnePåhlsson, Peter
By organisation
Clinical ChemistryFaculty of Health Sciences
In the same journal
Scandinavian Journal of Immunology
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 58 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf