title: Diesel exhaust particulate--exposed macrophages cause marked endothelial cell activation.
abstract: Exposure to air pollution containing diesel exhaust particulate (DEP) is linked to adverse cardiovascular events. This study tested the hypothesis that DEP not only causes direct endothelial cell injury, but also induces indirect endothelial cell activation via the release of soluble proinflammatory cytokines from macrophages. Human umbilical vein endothelial cells (HUVECs) and monocyte-derived macrophages (MDMs) were incubated with DEP (1-100 μg/ml; 24 h). Supernatants were analyzed for monocyte chemotactic protein (MCP)-1, IL6, IL8, and TNF-α. Indirect actions of DEP were investigated by incubating HUVECs with conditioned media from DEP-exposed MDMs in the presence and absence of the TNF-α inhibitor, etanercept. A modified Boyden chamber assay was used to determine whether HUVECs treated in this manner induced monocyte chemotaxis. Direct incubation with DEP induced a modest increase in MCP-1 concentration, but had no effect on IL-6 or IL-8 release from HUVECs. In contrast, direct treatment of MDMs with DEP had no effect on MCP-1, but elevated IL-8 and TNF-α concentrations. Incubation with conditioned media from DEP-exposed MDMs caused a dramatic amplification in MCP-1 and IL-6, but not IL-8, release from HUVECs. The potentiation of HUVEC activation was suppressed by TNF-α inhibition. MCP-1- and IL-6-containing HUVEC supernatants caused increased monocyte chemotaxis that was not inhibited by anti-MCP-1 antibodies. We conclude that DEP has only modest direct endothelial effects. In contrast, proinflammatory cytokines released from particle-laden MDMs appear to exacerbate endothelial activation after DEP exposure.
authors.affiliation: Centre for Cardiovascular Science, University of Edinburgh, Edinburgh, Scotland, UK. catherine.shaw@ed.ac.uk
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Shaw
authors.forename: Sarah
authors.initials: S
authors.lastname: Robertson
authors.forename: Mark R
authors.initials: MR
authors.lastname: Miller
authors.forename: Rodger
authors.initials: R
authors.lastname: Duffin
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Tabor
authors.forename: Ken
authors.initials: K
authors.lastname: Donaldson
authors.forename: David E
authors.initials: DE
authors.lastname: Newby
authors.forename: Patrick W F
authors.initials: PW
authors.lastname: Hadoke
mesh_terms: D002470:Cell Survival; D002633:Chemotaxis; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects; D008954:Models, Biological; D009000:Monocytes / Q000166:cytology; D010316:Particle Size; D010587:Phagocytosis; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D001335:Vehicle Emissions
chemical_list: D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D001335:Vehicle Emissions