Neutrophil-mediated damage to human vascular endothelium. Role of cytokine activation.
Cytokine activation of cultured human vascular endothelial cells renders them hyperadhesive for blood leukocytes. Co-incubation of freshly isolated, unstimulated human blood neutrophils with confluent cytokine-activated human endothelial monolayers for 90 minutes results in extensive endothelial detachment and destruction of monolayer integrity. In contrast, unactivated endothelial monolayers remain intact. Using this in vitro model, we have explored the neutrophil-effector mechanisms involved in this injury. Coincubation in the presence of a serine protease inhibitor (phenylmethylsulfonyl fluoride) or specific elastase inhibitors (Ala-Ala-Pro-Val-chloromethyl ketone or alpha-1-protease inhibitor) markedly diminished injury. In contrast, scavengers or inhibitors of oxygen-derived free radicals (superoxide dismutase, catalase, mannitol, or sodium azide) were not protective. Purified human neutrophil elastase mimicked the effect of the neutrophils suggesting a key role for elastase in the neutrophil-mediated injury in this model. Interfering with direct neutrophil-endothelial cell contact by interposing a microporous barrier insert prevented endothelial cell detachment. Furthermore, this neutrophil-mediated detachment could be inhibited with interleukin-8, an action correlated with a decrease in neutrophil adhesion to activated endothelial monolayers. By defining the role of endothelial activation in neutrophil-mediated injury, this in vitro model may provide useful insights into potential therapeutic interventions designed to prevent disruption of the endothelial barrier function.
Westlin
W F
WF
Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts.
Gimbrone
M A
MA
D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002450:Cell Communication / Q000187:drug effects*; D002478:Cells, Cultured; D010450:Endopeptidases / Q000502:physiology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000473:pathology* / Q000502:physiology; D006801:Humans; D007375:Interleukin-1 / Q000502:physiology*; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D014471:Umbilical Veins
D007375:Interleukin-1; D010450:Endopeptidases