Cytokine-treated human neutrophils contain inducible nitric oxide synthase that produces nitration of ingested bacteria.
Although the production of NO within rodent phagocytes is well-characterized, its production and function within human phagocytes are less clear. We show here that neutrophils within human buffy coat preparations stimulated with a mixture of interleukin 1, tumor necrosis factor alpha, and interferon gamma contain inducible NO synthase mRNA and protein, one of the enzymes responsible for NO production. The protein colocalizes with myeloperoxidase within neutrophil primary granules. Using an inhibitor of NO synthase, L-N-monomethyl arginine, we show that activity of this enzyme is required for the formation of nitrotyrosine around phagocytosed bacteria, most likely through the intermediate production of peroxynitrite, a reaction product of NO and superoxide anions.
Evans
T J
TJ
Department of Infections Diseases and Bacteriology, Royal Postgraduate Medical School, London, United Kingdom. tevans@rpms.ac.uk
Buttery
L D
LD
Carpenter
A
A
Springall
D R
DR
Polak
J M
JM
Cohen
J
J
D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001419:Bacteria / Q000276:immunology*; D001770:Blood Bactericidal Activity; D015153:Blotting, Western; D004790:Enzyme Induction; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D017403:In Situ Hybridization; D007371:Interferon-gamma / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology; D015846:Monokines / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D010587:Phagocytosis; D012333:RNA, Messenger / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D019323:omega-N-Methylarginine
D004791:Enzyme Inhibitors; D007375:Interleukin-1; D015846:Monokines; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D013481:Superoxides; D019323:omega-N-Methylarginine; C002744:3-nitrotyrosine; D014443:Tyrosine; D007371:Interferon-gamma; D001120:Arginine; D009195:Peroxidase; D019001:Nitric Oxide Synthase