title: A two-insult in vitro model of PMN-mediated pulmonary endothelial damage: requirements for adherence and chemokine release.
abstract: Lysophosphatidylcholines (lyso-PCs), generated during blood storage, are etiologic in a two-insult, sepsis-based model of transfusion-related acute lung injury (TRALI). Individually, endotoxin (LPS) and lyso-PCs prime but do not activate neutrophils (PMNs). We hypothesized that priming of PMNs alters their reactivity such that a second priming agent causes PMN activation and endothelial cell damage. PMNs were primed or not with LPS and then treated with lyso-PCs, and oxidase activation and elastase release were measured. For coculture experiments, activation of human pulmonary microvascular endothelial cells (HMVECs) was assessed by ICAM-1 expression and chemokine release. HMVECs were stimulated or not with LPS, PMNs were added, cells were incubated with lyso-PCs, and the number of viable HMVECs was counted. Lyso-PCs activated LPS-primed PMNs. HMVEC activation resulted in increased ICAM-1 and release of ENA-78, GRO alpha, and IL-8. PMN-mediated HMVEC damage was dependent on LPS activation of HMVECs, chemokine release, PMN adhesion, and lyso-PC activation of the oxidase. In conclusion, sequential exposure of PMNs to priming agents activates the microbicidal arsenal, and PMN-mediated HMVEC damage was the result of two insults: HMVEC activation and PMN oxidase assembly.
authors.affiliation: Bonfils Blood Center and Department of Pediatrics, University of Colorado School of Medicine, Denver, Colorado 80230, USA.
authors.forename: Travis H
authors.initials: TH
authors.lastname: Wyman
authors.forename: A Jason
authors.initials: AJ
authors.lastname: Bjornsen
authors.forename: David J
authors.initials: DJ
authors.lastname: Elzi
authors.forename: C Wayne
authors.initials: CW
authors.lastname: Smith
authors.forename: Kelly M
authors.initials: KM
authors.lastname: England
authors.forename: Marguerite
authors.initials: M
authors.lastname: Kelher
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Silliman
mesh_terms: D002448:Cell Adhesion; D002452:Cell Count; D002478:Cells, Cultured; D054360:Chemokine CXCL1; D054365:Chemokine CXCL5; D018925:Chemokines / Q000378:metabolism; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000378:metabolism; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000473:pathology* / Q000503:physiopathology*; D004789:Enzyme Activation; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D016209:Interleukin-8 / Q000031:analogs & derivatives* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008833:Microcirculation; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010088:Oxidoreductases / Q000378:metabolism; D010196:Pancreatic Elastase / Q000378:metabolism; D011652:Pulmonary Circulation
chemical_list: C475829:CXCL1 protein, human; C486263:CXCL5 protein, human; D054360:Chemokine CXCL1; D054365:Chemokine CXCL5; D018925:Chemokines; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D008244:Lysophosphatidylcholines; D018799:Intercellular Adhesion Molecule-1; D010088:Oxidoreductases; D010196:Pancreatic Elastase