{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,28]],"date-time":"2025-09-28T08:28:47Z","timestamp":1759048127288,"version":"3.38.0"},"reference-count":67,"publisher":"American Veterinary Medical Association (AVMA)","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["ajvr"],"published-print":{"date-parts":[[2002,3,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n\t\t\t\t<jats:p>\n\t\t\t\t\t<jats:bold>Objective<\/jats:bold>\u2014To develop an in vitro model of the bovine\nalveolar-capillary interface and to evaluate the roles of\ninterleukin-8 (IL-8) and platelet-activating factor (PAF)\nin neutrophil-mediated endothelial injury induced by\ninfection with <jats:italic>Mannheimia haemolytica<\/jats:italic>.<\/jats:p>\n\t\t\t\t<jats:p>\n\t\t\t\t\t<jats:bold>Sample Population<\/jats:bold>\u2014Cultured bovine pulmonary\nmicrovascular endothelial cells, freshly isolated\nbovine neutrophils, and monocyte-derived bovine\nmacrophages.<\/jats:p>\n\t\t\t\t<jats:p>\n\t\t\t\t\t<jats:bold>Procedure<\/jats:bold>\u2014A coculture system was developed in\nwhich endothelial cells were grown to confluence in\ntissue culture inserts, neutrophils were added to the\ninserts, and macrophages were added to tissue culture\nwells. <jats:italic>Mannheimia haemolytica<\/jats:italic>-derived\nlipopolysaccharide (LPS) or supernatant was added to\nactivate macrophages, and inhibitors of PAF or IL-8\nwere added to the insert. Endothelial cell cytotoxicity\nand permeability (ie, albumin leakage) and neutrophil\nactivation (ie, adhesion, degranulation [lactoferrin\nexpression], and superoxide production) were\nassessed.<\/jats:p>\n\t\t\t\t<jats:p>\n\t\t\t\t\t<jats:bold>Results<\/jats:bold>\u2014The addition of <jats:italic>M haemolytica-<\/jats:italic>derived LPS\nto bovine macrophages in the coculture system\nresulted in significant increases in endothelial cell\ncytotoxicity and permeability and neutrophil degranulation\nand adhesion. Inhibition of IL-8 reduced\nendothelial cell permeability and neutrophil degranulation\ninduced by exposure to <jats:italic>M haemolytica<\/jats:italic>-derived\nsupernatant, whereas inhibition of PAF decreased\nsuperoxide release by neutrophils.<\/jats:p>\n\t\t\t\t<jats:p>\n\t\t\t\t\t<jats:bold>Conclusions and Clinical Relevance<\/jats:bold>\u2014In vitro activation\nof bovine macrophages by <jats:italic>M haemolytica<\/jats:italic>derived\nLPS resulted in neutrophil activation and neutrophil-\nmediated endothelial damage. Neutrophilmediated\nendothelial injury and neutrophil degranulation\nwere, at least in part, mediated by IL-8, whereas\nPAF promoted superoxide release by neutrophils in\nthis in vitro system designed to mimic the in vivo\nevents that occur during the early stages of bovine\npneumonic pasteurellosis. 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