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1 Bonfils Blood Center and Departments of 2 Pediatrics and 3 Surgery, University of Colorado School of Medicine, Denver, Colorado 80230; and 4 Department of Pediatrics, Section of Leukocyte Biology, Baylor College of Medicine, Houston, Texas 77030
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
, 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.
neutrophils; endotoxin; lysophosphatidylcholines
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