An in vitro whole blood model to test the effects of different stimuli conditions on the release of myeloperoxidase and elastase by equine neutrophils.
Horses are particularly sensitive and exposed to excessive inflammatory responses evolving toward an important stimulation of polymorphonuclear neutrophils (PMNs). The aim of this work was to stimulate equine neutrophils in whole blood and to evaluate their response by measuring the release of total and active myeloperoxidase (MPO) and total elastase, considered as markers of neutrophil stimulation and degranulation. Because of the critical importance of the concomitant presence of LPS and TNF-α in equine pathological situations, we combined these two natural mediators to stimulate PMN and compared the response with those obtained after the PMN stimulation with each mediator used alone and well-known artificial stimulation systems such as 12-phorbol 13-myristate acetate (PMA) and the combination of cytochalasin B (CB) and N-formyl-methionyl-leucyl-phenylalanine (fMLP). All the activation systems, PMA, CB/fMLP, TNF-α, LPS and LPS/TNF-α, induced a significant release of total MPO in whole blood but only the combinations CB/fMLP and LPS/TNF-α significantly favored the release of active MPO. Regarding the total elastase, we did not observe a significant release in all the stimulated conditions except with PMA. It appears clearly that the choice of the neutrophil stimulation model is fundamental for the selection of potentially active pharmacological agents, especially on MPO activity.
Center for Oxygen Research and Development, Institute of Chemistry B6a, University of Liège, Sart Tilman, 4000 Liège, Belgium. J.ceusters@ulg.ac.be
Justine D
JD
Ceusters
Didier A
DA
Serteyn
Grégory
G
Minguet
Geoffroy
G
de la Rebière de Pouyade
Jennifer
J
Romainville
Ginette P
GP
Deby-Dupont
Ange A
AA
Mouithys-Mickalad
Thierry J
TJ
Franck
D000818:Animals; D015415:Biomarkers; D015971:Gene Expression Regulation, Enzymologic; D006736:Horses / Q000097:blood*; D008070:Lipopolysaccharides / Q000378:metabolism; D008954:Models, Biological; D009504:Neutrophils / Q000378:metabolism*; D010196:Pancreatic Elastase / Q000235:genetics / Q000378:metabolism*; D009195:Peroxidase / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D015415:Biomarkers; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D009195:Peroxidase; D010196:Pancreatic Elastase