Oxygen metabolites induced by phorbol myristate acetate increase lateral diffusion of wheat germ agglutinin-labeled glycoconjugates in human polymorphonuclear leukocytes

Inflammation ◽  
1990 ◽  
Vol 14 (6) ◽  
pp. 631-644 ◽  
Author(s):  
Birgitta Johansson ◽  
Karl -Eric Magnusson
Keyword(s):  
Phorbol Myristate Acetate ◽  
Wheat Germ Agglutinin ◽  
Polymorphonuclear Leukocytes ◽  
Wheat Germ ◽  
Lateral Diffusion ◽  
Myristate Acetate ◽  
Human Polymorphonuclear Leukocytes ◽  
Oxygen Metabolites

Lateral diffusion of plasma membrane receptors labelled with either platelet-derived growth factor (PDGF) or wheat germ agglutinin (WGA) in human polymorphonuclear leukocytes and fibroblasts

1989 ◽  
Vol 9 (1) ◽  
pp. 63-73 ◽  
Author(s):  
Pia Ljungquist ◽  
Åke Wasteson ◽  
Karl-Eric Magnusson
Keyword(s):  
Growth Factor ◽  
Wheat Germ Agglutinin ◽  
Polymorphonuclear Leukocytes ◽  
Wheat Germ ◽  
Human Fibroblasts ◽  
Normal Serum ◽  
Membrane Receptors ◽  
Platelet Derived Growth Factor ◽  
Pdgf Receptor ◽  
Human Polymorphonuclear Leukocytes

The aims of the present investigation were (a) to compare the lateral mobility of membrane receptors of human fibroblasts and polymorphonuclear leukocytes (PMNL) labelled with either platelet-derived growth factor (PDGF), or the lectin wheat germ agglutinin (WGA), and (b) to study effects of serum or PDGF on the mobility of these receptor molecules in human fibroblasts. Human foreskin fibroblasts (AG 1523) were grown on coverslips either under standard (10%) or under serum-free conditions yielding “normal” and “starved” cells, respectively. The receptor mobility was studied in response to exposure to PDGF, or serum, in short time or prolonged incubations. Human polymorphonuclear leukocytes (PMNL) were adhered to microscope slides by clotting drops of blood. They were stained with rhodaminated PDGF or fluoresceinated WGA. The diffusion of labelled receptors was assessed with fluorescence recovery after photobleaching (FRAP). It was found that (a) fibroblasts grown at normal serum concentration had a lower diffusion coefficient (D=3×10−10 cm2 s−1) for the PDGF-receptor and a slightly lower mobile fraction (R=60%) than starved cells (D=5×10−10 cm2s−1 and R=73%), (b) addition of serum to starved cells increased both D and R for the PDGF receptor to 12×10−10 cm2 s−1 and 96%, respectively, (c) a similar pattern was obtained for WGA-labelled glycoconjugates indicating general membrane effects of serum-induced cell stimulation, and (d) in PMNL the PDGF receptor displayed motility characteristics (D=3–4×10−10 cm2 s−1 and R=59%) similar to those in fibroblasts, possibly suggesting equivalent anchorage mechanisms in the membrane.

scholarly journals Effect of phorbol myristate acetate on the oxidative metabolism of human polymorphonuclear leukocytes

Blood ◽  
1976 ◽  
Vol 47 (4) ◽  
pp. 545-554 ◽  
Author(s):  
LR DeChatelet ◽  
PS Shirley ◽  
RB Jr Johnston
Keyword(s):  
Phorbol Myristate Acetate ◽  
Oxidative Metabolism ◽  
Polymorphonuclear Leukocytes ◽  
Time Course ◽  
Human Neutrophils ◽  
Hexose Monophosphate ◽  
Myristate Acetate ◽  
Bacterial Killing ◽  
Hexose Monophosphate Shunt ◽  
Human Polymorphonuclear Leukocytes

The addition of 0.1 mug/ml of phorbol myristate acetate (PMA) to a suspension of resting human neutrophils causes a marked stimulation of all aspects of cellular oxidative metabolism normally associated with phagocytosis. PMA induces a greatly increased rate of glucose oxidation via the hexose monophosphate shunt, increased production of superoxide anion and of hydrogen peroxide, increased cellular chemiluminescence, and increased iodination of protein material. The time course of hexose monophosphate shunt activation and of chemiluminescence are similar to those observed following phagocytosis of opsonized zymosan; the levels of activation achieved in all cases approximate those seen following phagocytosis. These phenomena are not simply reflections of altered cellular permeability, since PMA actually inhibits the uptake of radioactive 2-deoxyglucose and of uniformly labeled amino acids. The addition of PMA similarly inhibits the uptake of 14C-labeled bacteria, suggesting a competition between the effect of the chemical and the process of phagocytosis. These results suggest that PMA activates the cell in the same manner as does phagocytosis. This compound should provide a useful tool for elucidating the metabolic events underlying the phenomena of phagocytosis and bacterial killing by polymorphonuclear leukocytes.

scholarly journals Mechanisms of lysosomal enzyme release from human polymorphonuclear leukocytes. Effects of phorbol myristate acetate.

1975 ◽  
Vol 66 (3) ◽  
pp. 647-652 ◽  
Author(s):  
I M Goldstein ◽  
S T Hoffstein ◽  
G Weissmann
Keyword(s):  
Phorbol Myristate Acetate ◽  
Lysosomal Enzyme ◽  
Polymorphonuclear Leukocytes ◽  
Cytochalasin B ◽  
Enzyme Release ◽  
Myristate Acetate ◽  
Azurophil Granule ◽  
Cytoplasmic Microtubules ◽  
Specific Granule ◽  
Human Polymorphonuclear Leukocytes

PMA enhanced release of the azurophil granule enzyme, beta-glucuronidase, as well as lysozyme, from cytochalasin B-treated PMN's exposed to either zymosan particles or C5a. PMA was active at nanomolar concentrations, was not toxic to the cells, and was most effective when present for brief durations (0-1 min) before exposure of the cells to the stimuli. Beta-glucuronidase was not released in significant amounts from PMN's exposed to PMA alone, in the absence of stimuli such as zymosan or C5a. In contrast, only the specific granule enzyme, lysozyme, was released from unstimulated cells. Electron micrographs of cells exposed to PMA revealed an increase in the number of visible cytoplasmic microtubules as compared to control cells. Enhancement of lysosomal enzyme (beta-glucuronidase) release by PMA appears to be independent of effects on release of specific granule enzymes (lysozyme), but rather is likely due to PMA-induced elevations of cellular cGMP.

Generation of chemiluminescence by human neutrophils exposed to soluble stimuli of oxidative metabolism

1980 ◽  
Vol 30 (2) ◽  
pp. 385-390
Author(s):  
M A Westrick ◽  
P S Shirley ◽  
L R DeChatelet
Keyword(s):  
Bovine Serum Albumin ◽  
Phorbol Myristate Acetate ◽  
Polymorphonuclear Leukocytes ◽  
Reaction Medium ◽  
Human Neutrophils ◽  
Myristate Acetate ◽  
Secondary Interaction ◽  
Egg Albumin ◽  
Enhanced Chemiluminescence ◽  
Human Polymorphonuclear Leukocytes

The detection of chemiluminescence of human polymorphonuclear leukocytes activated by the nonparticulate stimulus phorbol myristate acetate required the presence of suitable substrate, such as protein or luminol, in the reaction medium. This substrate requirement was met by the addition of human serum or various proteins, such as bovine serum albumin, lysozyme, egg albumin, or immunoglobulin, to the reaction vial. Luminol, a chemiluminescent compound, could substitute for protein and markedly enhanced chemiluminescence by phorbol myristate acetate-induced and concanavalin A-induced polymorphonuclear leukocytes. From this work, it appears likely that soluble stimuli activate the polymorphonuclear leukocytes, but this activation, as measured by chemiluminescence, is not detectable in the absence of a secondary interaction with suitable components in the medium.

scholarly journals Effect of phorbol myristate acetate on the oxidative metabolism of human polymorphonuclear leukocytes

Blood ◽  
1976 ◽  
Vol 47 (4) ◽  
pp. 545-554 ◽  
Author(s):  
LR DeChatelet ◽  
PS Shirley ◽  
RB Jr Johnston
Keyword(s):  
Phorbol Myristate Acetate ◽  
Oxidative Metabolism ◽  
Polymorphonuclear Leukocytes ◽  
Time Course ◽  
Human Neutrophils ◽  
Hexose Monophosphate ◽  
Myristate Acetate ◽  
Bacterial Killing ◽  
Hexose Monophosphate Shunt ◽  
Human Polymorphonuclear Leukocytes

Abstract The addition of 0.1 mug/ml of phorbol myristate acetate (PMA) to a suspension of resting human neutrophils causes a marked stimulation of all aspects of cellular oxidative metabolism normally associated with phagocytosis. PMA induces a greatly increased rate of glucose oxidation via the hexose monophosphate shunt, increased production of superoxide anion and of hydrogen peroxide, increased cellular chemiluminescence, and increased iodination of protein material. The time course of hexose monophosphate shunt activation and of chemiluminescence are similar to those observed following phagocytosis of opsonized zymosan; the levels of activation achieved in all cases approximate those seen following phagocytosis. These phenomena are not simply reflections of altered cellular permeability, since PMA actually inhibits the uptake of radioactive 2-deoxyglucose and of uniformly labeled amino acids. The addition of PMA similarly inhibits the uptake of 14C-labeled bacteria, suggesting a competition between the effect of the chemical and the process of phagocytosis. These results suggest that PMA activates the cell in the same manner as does phagocytosis. This compound should provide a useful tool for elucidating the metabolic events underlying the phenomena of phagocytosis and bacterial killing by polymorphonuclear leukocytes.

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