Oxygenation activities of chicken polymorphonuclear leukocytes investigated by selective chemiluminigenic probes.
The redox metabolism of myeloperoxidase-deficient rooster (chicken) polymorphonuclear leukocytes (PMNL) was analyzed by differential chemiluminigenic probes. Chicken complement-opsonified zymosan, a phagocytosable particulate stimulus, and phorbol myristate acetate, a chemical stimulus, were used to activate the PMNL respiratory burst. The two probes used were luminol (5-amino-2,3-dihydro-1,4-phthalazinedione), a general probe of oxidase-peroxidase activities, and lucigenin (dimethylbiacridinium binitrate), a selective probe of oxidase activity. Rooster PMNLs yielded dimethylbiacridinum binitrate-dependent chemiluminescence (CL) comparable to those of myeloperoxidase-containing human PMNLs after stimulation with opsonified zymosan and to a lesser extent with phorbol myristate acetate. However, the luminol-dependent CL of opsonified zymosan or phorbol myristate acetate-stimulated rooster PMNLs were approximately two orders of magnitude lower than responses observed with human PMNLs. At physiologic pH, luminol is a highly sensitive, but not specific, probe of myeloperoxidase activity. Rooster erythrocytes yielded no CL with any of the probe-stimulus combinations described. Rooster PMNL viability and oxygen were required for CL. No strong correlation could be drawn between CL responses and eosinophil leukocyte concentration. The major conclusion is that rooster PMNLs, which do not have myeloperoxidase, present a significant and reproducible oxidative burst to chemical and particulate stimuli. Although lacking in peroxidase, rooster PMNLs can still present small luminol-dependent responses.
Department of Clinical Investigation, Brooke Army Medical Center, Fort Sam Houston, Texas 78234-6200, USA.
G A
GA
Merrill
R
R
Bretthauer
J
J
Wright-Hicks
R C
RC
Allen
D000166:Acridines / Q000494:pharmacology; D000818:Animals; D001799:Blood Sedimentation; D002645:Chickens / Q000378:metabolism*; D004912:Erythrocytes / Q000502:physiology; D006801:Humans; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D008165:Luminol / Q000494:pharmacology; D008297:Male; D009504:Neutrophils / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D009195:Peroxidase / Q000172:deficiency; D016897:Respiratory Burst / Q000502:physiology*
D000166:Acridines; D007202:Indicators and Reagents; C033472:10,10'-dimethyl-9,9'-biacridinium; D008165:Luminol; D009195:Peroxidase; D010100:Oxygen