Stimulus-dependent transduction mechanisms for nitric oxide release in human polymorphonuclear neutrophil leukocytes.
The production of nitric oxide (NO) may play an important role in functional responses of the human polymorphonuclear neutrophil granulocytes (PMNs). Others have described the presence of both an inducible, Ca2+-independent and a constitutionally expressed, Ca2+-dependent nitric oxide synthase (NOS) in human PMNs. However, the conditions for production and release of NO in human PMNs are still largely unknown. We assessed mechanisms for activation of NO release from human PMNs and particularly the dependence on extracellular and intracellular Ca2+. We addressed this question by applying a variety of agonists with known and differing mechanisms of activation in PMNs and measuring the released NO by two highly sensitive and specific real-time methods for detection of NO, the oxidation of oxyhemoglobin to methemoglobin and an electrochemical method. We found that human PMNs activated with the surface receptor-dependent agonist, N-formyl-methionyl-leucyl-phenylalanine (fMLP); the calcium ionophore, A23187; or the direct stimulator of protein kinase C, phorbol myristate acetate (PMA), produced NO which was inhibited by a specific NOS inhibitor, NG-monomethyl-L-arginine. The NO production induced by fMLP or A23187 was dependent on the presence of extracellular Ca2+, but this was not the case for PMA. The stimulatory effect of fMLP was almost completely inhibited by Bordetella pertussis toxin. These results indicate an NOS activity in purified human PMNs in vitro, and the transduction mechanisms for the agonists used show strong similarity with previously known pathways for other neutrophil functions.
Department of Hematology, Karolinska Institute at Huddinge University Hospital, Sweden.
G
G
Lärfars
H
H
Gyllenhammar
D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D013481:Superoxides; D009569:Nitric Oxide; D000001:Calcimycin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium