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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\u2013100 \u03bcg\/ml; 24 h). Supernatants were analyzed for monocyte chemotactic protein (MCP)\u20131, IL6, IL8, and TNF-\u03b1. Indirect actions of DEP were investigated by incubating HUVECs with conditioned media from DEP-exposed MDMs in the presence and absence of the TNF-\u03b1 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-\u03b1 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-\u03b1 inhibition. MCP-1\u2013 and IL-6\u2013containing HUVEC supernatants caused increased monocyte chemotaxis that was not inhibited by anti\u2013MCP-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.<\/jats:p>","DOI":"10.1165\/rcmb.2010-0011oc","type":"journal-article","created":{"date-parts":[[2010,8,6]],"date-time":"2010-08-06T22:50:44Z","timestamp":1281135044000},"page":"840-851","source":"Crossref","is-referenced-by-count":50,"title":["Diesel Exhaust Particulate\u2013Exposed Macrophages Cause Marked Endothelial Cell Activation"],"prefix":"10.1093","volume":"44","author":[{"given":"Catherine A.","family":"Shaw","sequence":"first","affiliation":[{"name":"Centre for Cardiovascular Science"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Robertson","sequence":"additional","affiliation":[{"name":"Centre for Cardiovascular Science"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark R.","family":"Miller","sequence":"additional","affiliation":[{"name":"Centre for Cardiovascular Science"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodger","family":"Duffin","sequence":"additional","affiliation":[{"name":"Centre for Inflammation Research, University of Edinburgh, Edinburgh, Scotland, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline M.","family":"Tabor","sequence":"additional","affiliation":[{"name":"Centre for Cardiovascular Science"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ken","family":"Donaldson","sequence":"additional","affiliation":[{"name":"Centre for Inflammation Research, University of Edinburgh, Edinburgh, Scotland, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David E.","family":"Newby","sequence":"additional","affiliation":[{"name":"Centre for Cardiovascular Science"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick W. 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