Article (Scientific journals)
Activation of equine neutrophils by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine induces a different response in reactive oxygen species production and release of active myeloperoxidase.
Franck, Thierry; Kohnen, Stéphane; de la Rebière de Pouyade, Geoffroy et al.
2009In Veterinary Immunology and Immunopathology
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Keywords :
Neutrophils; Equine; Stimulation; Oxidative activity; Myeloperoxidase
Abstract :
[en] Neutrophil (PMN) contribution to the acute inflammatory processes may lead to an excessive generation of reactive oxygen metabolites species (ROS) and secretion of granule enzymes. We compared the effects of either phorbol myristate acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP) in combination with a pre-treatment by cytochalasin B (CB) on the production of ROS and the release of total and active myeloperoxidase (MPO) by isolated equine PMNs. The ROS production was assessed by lucigenin dependent chemiluminescence (CL) and ethylene release by alpha-keto-gamma-methylthiobutyric acid (KMB) oxidation. In the supernatant of activated PMNs, total equine MPO was measured by ELISA and active MPO by the SIEFED (Specific Immunologic Extraction Followed by Enzymatic Detection) technique that allows for the study of the interaction of a compound directly with the enzyme. The stimulation of PMNs with CB-fMLP only modestly increased the release of MPO, but more than 70% of released MPO was active. PMA stimulation markedly increased the production of ROS and release of MPO, but more than 95% of released MPO was inactive. When PMNs were pre-incubated with superoxide dismutase (SOD) prior to PMA activation, the lucigenin enhanced CL, which is linked to the superoxide anion (O(2)(-)) production, was much more decreased than KMB oxidation, linked to the hydroxyl-like radical production. The addition of SOD prior to the activation of PMNs by PMA also limited the loss of the activity of released MPO. These results confirm the key role of O(2)(-) generation in the ROS cascade in PMN and reveal its critical role on MPO inactivation.
Research center :
CORD - Centre de l'Oxygène, Recherche et Développement - ULiège
Disciplines :
Veterinary medicine & animal health
Biochemistry, biophysics & molecular biology
Author, co-author :
Franck, Thierry  ;  Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Kohnen, Stéphane
de la Rebière de Pouyade, Geoffroy  ;  Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Deby, Ginette ;  Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Deby, Christiane ;  Centre Hospitalier Universitaire de Liège - CHU > Service administration des patients
Niesten, Ariane ;  Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Serteyn, Didier  ;  Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
Activation of equine neutrophils by phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine induces a different response in reactive oxygen species production and release of active myeloperoxidase.
Publication date :
2009
Journal title :
Veterinary Immunology and Immunopathology
ISSN :
0165-2427
eISSN :
1873-2534
Publisher :
Elsevier Science, Amsterdam, Netherlands
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 28 May 2009

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