A two-insult in vitro model of PMN-mediated pulmonary endothelial damage: requirements for adherence and chemokine release

2002 ◽  
Vol 283 (6) ◽  
pp. C1592-C1603 ◽  
Author(s):  
Travis H. Wyman ◽  
A. Jason Bjornsen ◽  
David J. Elzi ◽  
C. Wayne Smith ◽  
Kelly M. England ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
In Vitro Model ◽  
Cell Damage ◽  
Endothelial Damage ◽  
Microvascular Endothelial Cells ◽  
Blood Storage ◽  
Endothelial Cell Damage ◽  
Vitro Model ◽  
Lps Activation

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, GROα, 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.

scholarly journals Donor antibodies to HNA-3a implicated in TRALI reactions prime neutrophils and cause PMN-mediated damage to human pulmonary microvascular endothelial cells in a two-event in vitro model

Blood ◽  
2006 ◽  
Vol 109 (4) ◽  
pp. 1752-1755 ◽  
Author(s):  
Christopher C. Silliman ◽  
Brian R. Curtis ◽  
Patricia M. Kopko ◽  
Samina Y. Khan ◽  
Marguerite R. Kelher ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
In Vitro Model ◽  
Microvascular Endothelial Cells ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Biologic Response ◽  
Event Model ◽  
Microvascular Endothelial ◽  
Vitro Model ◽  
Related Mortality

Abstract Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related mortality. Antibodies to HNA-3a are commonly implicated in TRALI. We hypothesized that HNA-3a antibodies prime neutrophils (PMNs) and cause PMN-mediated cytotoxicity through a two-event pathogenesis. Isolated HNA-3a+ or HNA-3a− PMNs were incubated with plasma containing HNA-3a antibodies implicated in TRALI, and their ability to prime the oxidase was measured. Human pulmonary microvascular endothelial cells (HMVECs) were activated with endotoxin or buffer, HNA-3a+ or HNA-3a− PMNs were added, and the coculture was incubated with plasma ± antibodies to HNA-3a. PMN-mediated damage was measured by counting viable HMVECs/mm2. Plasma containing HNA-3a antibodies primed the fMLP-activated respiratory burst of HNA-3a+, but not HNA-3a−, PMNs and elicited PMN-mediated damage of LPS-activated HMVECs when HNA-3a+, but not HNA-3a−, PMNs were used. Thus, antibodies to HNA-3a primed PMNs and caused PMN-mediated HMVEC cytotoxicity in a two-event model identical to biologic response modifiers implicated in TRALI.

A New in Vitro Model for Agonist-Induced Communication between Microvascular Endothelial Cells

2000 ◽  
Vol 60 (3) ◽  
pp. 222-231 ◽  
Author(s):  
Yves Ouellette ◽  
Darcy Lidington ◽  
Christian G. Naus ◽  
Karel Tyml
Keyword(s):  
Endothelial Cells ◽  
In Vitro Model ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
Vitro Model
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