scholarly journals Staurosporine inhibits the respiratory burst and induces exocytosis in human neutrophils

1989 ◽  
Vol 264 (3) ◽  
pp. 879-884 ◽  
Author(s):  
B Dewald ◽  
M Thelen ◽  
M P Wymann ◽  
M Baggiolini
Keyword(s):  
Vitamin B12 ◽  
Protein Phosphorylation ◽  
Phorbol Myristate Acetate ◽  
Respiratory Burst ◽  
Cytochalasin B ◽  
Binding Protein ◽  
Human Neutrophils ◽  
Myristate Acetate ◽  
Chemotactic Peptides ◽  
Protein Kinase C Inhibitor

The protein kinase C inhibitor staurosporine influenced in different ways the functions of human neutrophils. Staurosporine prevented the enhanced protein phosphorylation in phorbol ester- and N-formylmethyionyl-leucylphenylalanine (fMLP)-stimulated cells, and was a powerful inhibitor of the respiratory burst induced by phorbol myristate acetate [IC50 (concentration causing 50% inhibition) 17 nM] and the chemotactic peptides fMLP and C5a (IC50 24 nM). It did not alter, however, the superoxide production by cell-free preparations of NADPH oxidase. Staurosporine had no effect on agonist-dependent changes in cytosolic free Ca2+ and exocytosis of specific and azurophil granules, and showed only a slight inhibition of the release of vitamin B12-binding protein induced by phorbol myristate acetate (decreased by 40% at 200 nM). On the other hand, staurosporine also exhibited neutrophil-activating properties: it induced the release of gelatinase (from secretory vesicles) and vitamin-B12-binding protein (from specific granules). These effects were protracted, concentration-dependent, insensitive to Ca2+ depletion, and strongly enhanced by cytochalasin B. Staurosporine, however, did not induce the release of beta-glucuronidase or elastase (from azurophil granules). Except for the sensitivity to cytochalasin B, these properties suggest a similarity between the exocytosis-inducing actions of staurosporine and PMA. The results obtained with staurosporine provide further evidence that different signal-transduction processes are involved in neutrophil activation, and suggest that protein phosphorylation is required for the induction of the respiratory burst, but not for exocytosis.

Suppressive action of cobalt on exocytosis and respiratory burst in neutrophils

1989 ◽  
Vol 257 (5) ◽  
pp. C859-C864 ◽  
Author(s):  
J. G. Elferink ◽  
M. Deierkauf
Keyword(s):  
Plasma Membrane ◽  
Phorbol Myristate Acetate ◽  
Respiratory Burst ◽  
Cytochalasin B ◽  
Superoxide Production ◽  
Co2 Activation ◽  
Chemotactic Peptide ◽  
Myristate Acetate ◽  
Intracellular Target ◽  
Suppressive Action

Activation of exocytosis and respiratory burst in rabbit neutrophils by the chemotactic peptide fMet-Leu-Phe is inhibited by Co2+. Inhibition is antagonized by extracellular Ca2+ and is dependent on the time of preincubation of cells with Co2+ before addition of activator. Co2+ inhibits the enhancement of 45Ca association that occurs during activation with fMet-Leu-Phe. Interference with Ca2+ translocation across the plasma membrane by Co2+ is probably not the cause of inhibition of neutrophil activation, because activation in the absence of extracellular Ca2+ is inhibited by Co2+. Activation of neutrophils by phorbol myristate acetate is inhibited at higher Co2+ concentrations than activation by fMet-Leu-Phe. Inhibition of the superoxide production by Co2+ occurs both in the presence or in the absence of cytochalasin B. Fluorescence of neutrophils loaded with quin2 is diminished by Co2+, indicating that Co2+ had entered into the cytoplasm. The results are compatible with the view that Co2+ inhibits exocytosis and respiratory burst in neutrophils by an interaction with a Ca2+-dependent intracellular target.

scholarly journals Chemotactic factor enhancement of superoxide release from fluoride and phorbol myristate acetate stimulated neutrophils

Blood ◽  
1981 ◽  
Vol 58 (1) ◽  
pp. 129-134 ◽  
Author(s):  
D English ◽  
JS Roloff ◽  
JN Lukens
Keyword(s):  
Phorbol Myristate Acetate ◽  
Normal Serum ◽  
Human Neutrophils ◽  
Chemotactic Peptide ◽  
Myristate Acetate ◽  
Chemotactic Peptides ◽  
Extracellular Glucose ◽  
Increased Activity ◽  
Generating System ◽  
Superoxide Release

Abstract Human neutrophils exposed to chemotactic concentrations of zymosan- activated serum (ZAS) and a formylated chemotactic peptide (FMLP, 10(- 7)--10(-9) M) were markedly enhanced in their ability to generate superoxide (O2-) upon stimulation with either sodium fluoride or phorbol myristate acetate (PMA). For both fluoride and PMA, enhancement was characterized by a decrease in the lag from stimulation to initiation of superoxide release and by an increase in the rate of superoxide generation--representing faster activation and increased activity of O2- generating enzyme, respectively. Chemotactic concentrations of casein, normal serum, and casein-treated serum enhanced the activity, but not the rate of activation, of the fluoride- stimulated superoxide generating system. This effect on activity was not so impressive as that obtained with FMLP or ZAS. The mechanisms by which FMLP enhanced responsiveness to fluoride and PMA were found to be different. Optimal enhancement for fluoride-stimulated responses required extracellular Ca++. Extracellular glucose, but not extracellular Ca++, was required for enhancement of FMLP of PMA- stimulated responses. A similar glucose requirement could not be demonstrated for chemotactic peptide enhancement of the superoxide- generating system stimulated by fluoride. Fluoride and PMA apparently activate the neutrophil O2- generating enzyme by pathways that are not identical. However, responsiveness of the enzyme to both agents is susceptible to modulation by cellular responses to chemotactic peptides.

Effect of phorbol myristate acetate and the chemotactic peptide fNLPNTL on shape and movement of human neutrophils

1987 ◽  
Vol 88 (3) ◽  
pp. 399-406 ◽  
Author(s):  
F.J. Roos ◽  
A. Zimmermann ◽  
H.U. Keller
Keyword(s):  
Phorbol Myristate Acetate ◽  
Human Neutrophils ◽  
Chemotactic Peptide ◽  
Myristate Acetate ◽  
Locomotor Apparatus ◽  
Kinase C ◽  
Motile Cells ◽  
Chemotactic Peptides ◽  
Intracellular Organelles ◽  
Stimulation Of

The results show that the distinct types of shape produced by phorbol myristate acetate (PMA) and by chemotactic peptides (fNLPNTL) are associated with distinct types of neutrophil movement. Whereas the chemotactic peptide can induce front-tail polarity characterized by an expanding front, a contracted tail and preferential unidirectional movements of intracellular organelles, PMA can only elicit non-polar movements characterized by random formation and retraction of projections all over the surface, intracellular movements of organelles being ill-defined and changing in direction. Combined stimulation of human neutrophils with PMA and fNLPNTL results in a suppression of peptide-induced polarity and the formation of non-polar motile cells resembling those stimulated with PMA alone. The results suggest that the diacylglycerol-protein kinase C pathway may be instrumental in transducing or modulating signals to the locomotor apparatus of the cell. PMA-treated cells are, however, still capable of developing directional responses when appropriately stimulated. The findings lead to the hypothesis that distinct types of neutrophil movements are preferentially associated with distinct functions.

scholarly journals Chemotactic factor enhancement of superoxide release from fluoride and phorbol myristate acetate stimulated neutrophils

Blood ◽  
1981 ◽  
Vol 58 (1) ◽  
pp. 129-134 ◽  
Author(s):  
D English ◽  
JS Roloff ◽  
JN Lukens
Keyword(s):  
Phorbol Myristate Acetate ◽  
Normal Serum ◽  
Human Neutrophils ◽  
Chemotactic Peptide ◽  
Myristate Acetate ◽  
Chemotactic Peptides ◽  
Extracellular Glucose ◽  
Increased Activity ◽  
Generating System ◽  
Superoxide Release

Human neutrophils exposed to chemotactic concentrations of zymosan- activated serum (ZAS) and a formylated chemotactic peptide (FMLP, 10(- 7)--10(-9) M) were markedly enhanced in their ability to generate superoxide (O2-) upon stimulation with either sodium fluoride or phorbol myristate acetate (PMA). For both fluoride and PMA, enhancement was characterized by a decrease in the lag from stimulation to initiation of superoxide release and by an increase in the rate of superoxide generation--representing faster activation and increased activity of O2- generating enzyme, respectively. Chemotactic concentrations of casein, normal serum, and casein-treated serum enhanced the activity, but not the rate of activation, of the fluoride- stimulated superoxide generating system. This effect on activity was not so impressive as that obtained with FMLP or ZAS. The mechanisms by which FMLP enhanced responsiveness to fluoride and PMA were found to be different. Optimal enhancement for fluoride-stimulated responses required extracellular Ca++. Extracellular glucose, but not extracellular Ca++, was required for enhancement of FMLP of PMA- stimulated responses. A similar glucose requirement could not be demonstrated for chemotactic peptide enhancement of the superoxide- generating system stimulated by fluoride. Fluoride and PMA apparently activate the neutrophil O2- generating enzyme by pathways that are not identical. However, responsiveness of the enzyme to both agents is susceptible to modulation by cellular responses to chemotactic peptides.

scholarly journals Phosphorylation of the subunits of cytochrome b-245 upon triggering of the respiratory burst of human neutrophils and macrophages

1988 ◽  
Vol 252 (3) ◽  
pp. 901-904 ◽  
Author(s):  
R C Garcia ◽  
A W Segal
Keyword(s):  
Cytochrome B ◽  
Phorbol Myristate Acetate ◽  
Respiratory Burst ◽  
Time Course ◽  
Autosomal Recessive ◽  
Beta Subunit ◽  
Human Neutrophils ◽  
Granulomatous Disease ◽  
Myristate Acetate ◽  
Oxidase System

Cytochrome b-245, the only clearly identified component of the microbicidal oxidase system of phagocytes, is a heterodimer consisting of a 23 kDa (alpha) and a 76-92 kDa (beta) subunit. This study was conducted to examine whether, in common with a number of proteins, the subunits of the cytochrome were phosphorylated upon activation of the oxidase. Both subunits were phosphorylated after activation of neutrophils or macrophages with phorbol myristate acetate or a phagocytic stimulus, although the time course of this process did not parallel that of the oxidase. Phosphorylation of these proteins was normal in cells from two patients with autosomal recessive chronic granulomatous disease, in whom phosphorylation of a 47 kDa protein is defective.

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