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Mast cell granules potentiate endotoxin‐induced interleukin‐6 production by endothelial cells

Mast cells are constituent cells of vascular tissue and their numbers are increased in atherosclerotic vessels. To gain insight into the role of mast cells in vascular inflammation, the effect of mast cell granules (MCG) on endothelial cell production of interleukin‐6 (IL‐6) was examined. Human umbilical vein endothelial cells (HUVEC) were cultured with lipopolysaccharide (LPS) in the presence or absence of rat peritoneal MCG and IL‐6 production was assayed by enzymelinked immunosorbent assay. The interaction of MCG with HUVEC in culture was examined by electron microscopy (EM). The EM studies revealed that MCG are internalized by HUVEC and appear intact even after 24 h in culture. Unactivated HUVEC produced little or no IL‐6 either in the presence or absence of MCG. Treatment of HUVEC with LPS stimulated IL‐6 production in a dose‐ and time‐dependent fashion. Addition of MCG to LPS‐activated HUVEC resulted in the potentiation of IL‐6 production at all LPS doses. MCG‐induced enhancement of IL‐6 production was evident even at a mast cell‐to‐endothelial cell ratio of 1:32. The enhancement of IL‐6 production by MCG was also seen when tumor necrosis factor α was used as an activator. Although potentiation was evident when MCG were added 6 h before or after LPS stimulation, the maximum effect was noted when MCG and LPS were added simultaneously. MCG‐mediated enhancement of IL‐6 production was abrogated by pretreating MCG with protease inhibitors. Although MCG proteases potentiate IL‐6 production by HUVEC, they do not degrade secreted IL‐6. These results demonstrate that MCG interact with endothelial cells and modulate the production of an important inflammatory cytokine. J. Leukoc. Biol. 62: 210–216; 1997.