title: Tumour necrosis factor-alpha induces neutrophil-mediated injury of cultured human endothelial cells.
abstract: We investigated the ability of TNF-alpha to mediate damage of endothelial cells in the presence of neutrophils, by measuring detachment of cultured human umbilical vein endothelial cells (HUVEC). Endothelial cell detachment was increased from 5% to about 75% by the presence of 1-10 ng/ml TNF-alpha during incubation with neutrophils, whereas negligible endothelial cell lysis was observed as measured by 51Cr release. TNF-alpha was compared with the cytokines IL-1 alpha and IFN-gamma and with PMA and LPS. Both TNF-alpha and PMA appeared to be strong triggers for neutrophil-induced endothelial cell detachment, whilst reduced injury was seen after addition of IL-1 alpha and LPS. IFN-gamma did not induce endothelial cell detachment, but potentiated the effect of both TNF-alpha and IL-1 alpha. TNF-alpha-induced endothelial cell detachment was neutrophil dependent, since pre-incubation of neutrophils, but not pre-incubation of endothelial cells with TNF-alpha, caused endothelial cell detachment. Thus, TNF-alpha-induced increase in neutrophil-adhesiveness of HUVEC was found not to be essential for endothelial damage. Pre-incubation of neutrophils in suspension with TNF-alpha induced rapid activation, followed by nearly complete deactivation of neutrophils, as measured by their capacity to induce detachment of endothelial cells after removal of TNF-alpha. These results indicate that local presence of TNF-alpha might be critical in tissue or organ damage during early, neutrophil-mediated inflammatory processes, independent of enhanced adhesiveness of endothelium for neutrophils.
authors.affiliation: Department of Surgery, University of Limburg, Maastricht, The Netherlands.
authors.forename: E J
authors.initials: EJ
authors.lastname: von Asmuth
authors.forename: J F
authors.initials: JF
authors.lastname: Leeuwenberg
authors.forename: C J
authors.initials: CJ
authors.lastname: van der Linden
authors.forename: W A
authors.initials: WA
authors.lastname: Buurman
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000473:pathology*; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D007375:Interleukin-1; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate