title: Human monocytes activate porcine endothelial cells, resulting in increased E-selectin, interleukin-8, monocyte chemotactic protein-1, and plasminogen activator inhibitor-type-1 expression.
abstract: Monocytes (Mo) are thought to be important effector cells in early xenograft rejection. Effects of Mo-endothelial cell (EC) interactions on EC activation in vitro were studied by coculturing human Mo or human monocytoid cell lines, U937 and THP-1, with porcine EC. Without preactivation, U937 cells and Mo induced mRNA for the EC-specific adhesion receptor, E-selectin, expressed only on activated cells, after 2 hr. Surface protein was maximal when equal numbers of EC and Mo were cocultured. Increased mRNA expression of the chemokines, interleukin-8 and monocyte chemotactic protein-1, and the antifibrinolytic protein plasminogen activator inhibitor type-1, confirmed EC activation. Like E-selectin, plasminogen activator inhibitor type-1 mRNA was rapidly induced and returned to baseline after 24 hr, whereas chemokine gene expression was slower and more prolonged. Interleukin-1 receptor antagonist failed to modulate induction of E-selectin. Soluble tumor necrosis factor (TNF) alpha receptor inhibited E-selectin induced by TNF alpha, but not by U937 cells, and mRNA and protein on EC in Mo-EC mixtures cocultured at 1:1 ratios were not significantly reduced. The TNF alpha inhibitor did reduce E-selectin expression (30-40%), as well as induced chemokine gene expression (80%), at higher Mo-EC ratios. Despite this, minimal TNF alpha was detectable in supernatants. These results, along with the transwell experiments that confirmed a requirement for Mo-EC contact, suggest that membrane-bound TNF alpha may be involved. Thus, Mo-EC interactions in the porcine to human combination activated several EC functions, suggesting that initial Mo contact with the vessel wall of a xenogeneic graft may promote leukocyte recruitment, inflammation, and maintenance of thrombus, resulting in eventual organ destruction.
authors.affiliation: Department of Surgery, Deaconess Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: M T
authors.initials: MT
authors.lastname: Millan
authors.forename: C
authors.initials: C
authors.lastname: Geczy
authors.forename: K M
authors.initials: KM
authors.lastname: Stuhlmeier
authors.forename: D J
authors.initials: DJ
authors.lastname: Goodman
authors.forename: C
authors.initials: C
authors.lastname: Ferran
authors.forename: F H
authors.initials: FH
authors.lastname: Bach
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000276:immunology; D002450:Cell Communication / Q000276:immunology; D002460:Cell Line; D018932:Chemokine CCL2 / Q000096:biosynthesis*; D018920:Coculture Techniques; D019040:E-Selectin / Q000096:biosynthesis*; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D007375:Interleukin-1 / Q000502:physiology; D016209:Interleukin-8 / Q000096:biosynthesis*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008262:Macrophage Activation; D009000:Monocytes / Q000276:immunology*; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis*; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology
chemical_list: D018932:Chemokine CCL2; D019040:E-Selectin; D007375:Interleukin-1; D016209:Interleukin-8; D017395:Plasminogen Activator Inhibitor 1; D014409:Tumor Necrosis Factor-alpha