scholarly journals MECHANISMS OF LYSOSOMAL ENZYME RELEASE FROM HUMAN LEUKOCYTES

1973 ◽  
Vol 58 (1) ◽  
pp. 27-41 ◽  
Author(s):  
Robert B. Zurier ◽  
Sylvia Hoffstein ◽  
Gerald Weissmann
Keyword(s):  
Plasma Membrane ◽  
Lysosomal Enzyme ◽  
Immune Complexes ◽  
Cyclic Nucleotides ◽  
Lysosomal Enzymes ◽  
Plasma Membranes ◽  
Prostaglandin E ◽  
Enzyme Release ◽  
Human Leukocytes ◽  
Oxidation Of Glucose

In order to study mechanisms underlying selective enzyme release from human leukocytes during phagocytosis, the effects were studied of compounds which affect microtubule integrity or the accumulation of cyclic nucleotides. Human leukocytes selectively extrude lysosomal enzymes (ß-glucuronidase) from viable cells during phagocytosis of zymosan or immune complexes, or upon encounter with immune complexes dispersed along a non-phagocytosable surface such as a millipore filter. In each circumstance, lysosomal enzyme release was reduced by previous treatment of cells with pharmacological doses of drugs which disrupt microtubules (e.g. 10-3–10-5 M colchicine) or with agents which affect accumulation of adenosine 3'5'-monophosphate (cAMP) (e.g. 10-3 M cyclic nucleotides and 2.8 x 10-4–2.8 x 10-6 M prostaglandin E (PGE) and A (PGA) compounds). Preincubation of cells with 5 µg/ml cytochalasin B resulted in complete inhibition of zymosan ingestion, but not of adherence of zymosan particles to plasma membranes or selective enzyme release. In this system, in which enzyme release was independent of particle uptake, preincubation of cells with colchicine, vinblastine, dibutyryl cAMP, or PGE1 also reduced extrusion of lysosomal enzymes. When cell suspensions were incubated with membrane-lytic crystals of monosodium urate (MSU), cytoplasmic as well as lysosomal enzymes were released with subsequent death of the cells. However, enzyme release followed phagocytosis of crystals (as measured by enhanced C-1 oxidation of glucose) and was due to "perforation from within" of the lysosomal membrane, rather than lysis by crystals of the plasma membrane. Enzyme release after MSU ingestion was also reduced when cells were treated with pharmacological doses of the test agents. When cells were killed by Triton X-100, acting on the plasma membrane, C-1 oxidation of glucose was abolished and enzyme release could not be inhibited pharmacologically. These observations suggest that lysosomal enzyme release from human phagocytes can be an active process which accompanies plasma membrane stimulation, is independent of cell death, and may be controlled by cyclic nucleotides and agents which affect microtubules.

MECHANISMS OF LYSOSOMAL ENZYME RELEASE FROM LEUKOCYTES EXPOSED TO IMMUNE COMPLEXES AND OTHER PARTICLES

1971 ◽  
Vol 134 (3) ◽  
pp. 149-165 ◽  
Author(s):  
Gerald Weissmann ◽  
Robert B. Zurier ◽  
Paul J. Spieler ◽  
Ira M. Goldstein
Keyword(s):  
Immune Complexes ◽  
Lysosomal Enzymes ◽  
Tris Buffer ◽  
Selective Absorption ◽  
Enzyme Release ◽  
Acid Hydrolases ◽  
Lysosomal Hydrolases ◽  
Fresh Serum ◽  
Inert Particles ◽  
Oxidation Of Glucose

Human PMN release lysosomal enzymes (ß-glucuronidase, acid phosphatase) when exposed to immune complexes, but do not release cytoplasmic LDH. The cells remain viable, and failure of LDH to appear in supernatants is not due to selective absorption or inactivation. Release of enzymes is not due to platelet contamination and is only partially enhanced by fresh serum. The selective release of lysosomal enzymes after uptake of complexes resembles that induced by inert particles of zymosan, and can be distinguished from the concurrent release of all enzymes after cell death induced by membrane-lytic crystals of MSU. Uptake of complexes, zymosan, or MSU particles is accompanied by concomitant increases in C-1 oxidation of glucose. Although MSU-induced damage can be retarded by the presence of Tris buffer, immune complexes and zymosan selectively release lysosomal hydrolases in the presence or absence of Tris buffer. Agents which elevate the level, within cells, of cAMP (PGE1, theophylline, 2-CA) and cAMP itself inhibit the selective extrusion of acid hydrolases from leukocytes without affecting the viability of cells. Leukocytes may respond to immune particles by regurgitating a portion of their lysosomal hydrolases during phagocytosis.

The Induction of Lysosomal Enzyme Release from Leucocytes of Normal and Emphysematous Subjects and the Effects of Tobacco Smoke upon Phagocytosis

1980 ◽  
Vol 58 (5) ◽  
pp. 403-409 ◽  
Author(s):  
D. C. S. Hutchison ◽  
R. Desai ◽  
D. Bellamy ◽  
H. Baum
Keyword(s):  
Cigarette Smoke ◽  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Soluble Fraction ◽  
Normal Subjects ◽  
Polymorphonuclear Leucocytes ◽  
Enzyme Release ◽  
Elastic Tissue ◽  
Latex Particles ◽  
Respiratory Inhibitor

1. The lysosomal enzymes of circulating polymorphonuclear leucocytes contain a potent elastase; release of this enzyme within the lung is thought to be responsible for the destruction of elastic tissue in pulmonary emphysema. 2. The release of lysosomal enzymes from blood leucocytes of normal and emphysematous subjects during phagocytosis of particulate material was studied In vitro. Acid phosphatase and acid ribonuclease were used as markers of lysosomal enzyme release, no sufficiently sensitive assay for elastase being available. Cigarette smoke was separated into ‘particulate’ and ‘soluble’ fractions. In a preliminary study, the particulate fraction stimulated enzyme release; in the experiments reported here, latex particles were used to produce this effect. 3. Approximately one-third of the total lysosomal enzyme content was released to the exterior of the cell during phagocytosis of latex particles. In this respect there was no difference between normal and emphysematous subjects. 4. The effects of the non-particulate soluble fraction of cigarette smoke on phagocytosis-induced enzyme release were studied. This fraction inhibited enzyme release from polymorphonuclear leucocytes of normal subjects but not from those of emphysematous patients. When the ‘cigarette-smoke solution’ was replaced by the respiratory inhibitor, antimycin A, a similar inhibition of enzyme release occurred. The inhibition of phagocytosis in cells of normal subjects is presumed to be due to a respiratory inhibitor such as carbon monoxide in the soluble fraction of the smoke. We postulate that the polymorphonuclear leucocytes of emphysematous patients are adapted to hypoxic conditions so that inhibition of enzyme release does not occur.

The interaction of human eosinophils and neutrophils with non-phagocytosable surfaces: a model for studying cell-mediated immunity in schistosomiasis

Parasitology ◽  
1980 ◽  
Vol 80 (3) ◽  
pp. 525-537 ◽  
Author(s):  
Audrey M. Glauert ◽  
Rhonda C. Oliver ◽  
Kareen J. I. Thorne
Keyword(s):  
Plasma Membrane ◽  
Lysosomal Enzymes ◽  
Cell Types ◽  
Cell Mediated Immunity ◽  
Enzyme Release ◽  
Basic Information ◽  
Intimate Contact ◽  
Effector Cells ◽  
Wide Range ◽  
Direct Fusion

SummaryA model has been developed to simulate the surface of an antibody-coated schistosomulum. It consists of a layer of agar, containing antigen (tetanus toxoid) and a chemotactic factor (ECF). Some layers were coated with human anti-tetanus immunoglobulin. The mode of adherence of human eosinophils and neutrophils to these agar layers and the subsequent degranulation of the cells exactly paralleled the interaction of these cell types with antibody-coated schistosomula ofSchistosoma mansoni. In particular, eosinophils made much more intimate contact than did neutrophils, and lysosomal enzymes were secreted extracellularly by direct fusion of granules with the plasma membrane of the cell. Biochemical evidence was also obtained for the secretion of enzymes during degranulation and the rate of enzyme release was found to be enhanced in the presence of specific antibody. This model, non-phagocytosable surface has the potential to provide basic information on the mode of action of effector cells in cell-mediated cytotoxic reactions against a wide range of parasites by incorporation of different factors into the agar layers.

Dissociation of phagocytosis, metabolic stimulation and lysosomal enzyme release in human leukocytes

1976 ◽  
Vol 6 (1-3) ◽  
pp. 256-259 ◽  
Author(s):  
Dirk Roos ◽  
Ira M. Goldstein ◽  
Howard B. Kaplan ◽  
Gerald Weissmann
Keyword(s):  
Lysosomal Enzyme ◽  
Enzyme Release ◽  
Human Leukocytes ◽  
Metabolic Stimulation

scholarly journals Role of microtubule assembly in lysosomal enzyme secretion from human polymorphonuclear leukocytes. A reevaluation.

1977 ◽  
Vol 73 (1) ◽  
pp. 242-256 ◽  
Author(s):  
S Hoffstein ◽  
I M Goldstein ◽  
G Weissmann
Keyword(s):  
Plasma Membrane ◽  
Dose Response ◽  
Lysosomal Enzyme ◽  
Polymorphonuclear Leukocytes ◽  
Cytochalasin B ◽  
Microtubule Assembly ◽  
Enzyme Release ◽  
Thin Sections ◽  
Human Polymorphonuclear Leukocytes ◽  
In Cells

The dose-related inhibition by colchicine of both lysosomal enzyme release and microtubule assembly was studied in human polymorphonuclear leukocytes (PMN) exposed to the nonphagocytic stimulus, zymosan-treated serum (ZTS). Cells were pretreated with colchicine (60 min, 37 degrees C) with or without cytochalasin B (5 microng/ml, 10 min) and then stimulated with ZTS (10%). Microtubule numbers in both cytochalasin B-treated and untreated PMN were increased by stimulation and depressed below resting levels in a dose-response fashion by colchicine concentrations above 10(-7) M. These concentrations also inhibited enzyme release in a dose-response fashion although the inhibition of microtubule assembly was proportionately greater than the inhibition of enzyme release. Other aspects of PMN morphology were affected by colchicine. Cytochalasin B-treated PMN were rounded, and in thin sections the retracted plasma membrane appeared as invaginations oriented toward centrally located centrioles. Membrane invaginations were restricted to the cell periphery in cells treated with inhibitory concentrations of colchicine, and the centrioles and Golgi apparatus were displaced from their usual position. After stimulation and subsequent degranulation, the size and number of membrane invaginations greatly increased. They remained peripheral in cells pretreated with greater than 10(-7) M colchicine but were numerous in the pericentriolar region in cells treated with less than 10(-7) M. Similarly, untreated PMN that were permitted to phagocytose immune precipitates had many phagosomes adjacent to the centriole. After colchicine treatment, phagosomes were distributed randomly, without any preferential association with the centrioles. These data suggest that microtubules are involved in maintaining the internal organization of cells and the topologic relationships between organelles and the plasma membrane.

scholarly journals HORMONAL CONTROL OF LYSOSOMAL ENZYME RELEASE FROM HUMAN NEUTROPHILS

1974 ◽  
Vol 139 (6) ◽  
pp. 1395-1414 ◽  
Author(s):  
L. J. Ignarro ◽  
T. F. Lint ◽  
W. J. George
Keyword(s):  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Hormonal Control ◽  
Human Neutrophils ◽  
Cell Contact ◽  
Enzyme Release ◽  
Lactate Dehydrogenase Activity ◽  
Particle Uptake

The purpose of this investigation was to examine the effects of autonomic neurohormones, cyclic nucleotides, and related agents on the immunologic discharge of lysosomal enzymes from, and phagocytosis by, purified human neutrophils. In order to discern the possible intracellular mechanisms by which certain neurohormones influence neutrophil function, the concentrations of cyclic AMP and cyclic GMP in neutrophils were assessed during cell contact with phagocytizable particles and autonomic agents. The model system employed for study was the interaction of purified human neutrophils with rheumatoid arthritic (RA) serum-treated zymosan particles at 37°C in a neutral, balanced salt solution containing glucose. Neutrophils ingested the particles and discharged ß-glucuronidase but not lactate dehydrogenase activity during 30 min of incubation. Treatment of zymosan particles with RA serum was more effective than treatment with normal serum with regard to the extent of both particle uptake and lysosomal enzyme release. During contact of neutrophils with RA serum-treated zymosan particles epinephrine, isoproterenol, and cyclic AMP inhibited both particle ingestion and ß-glucuronidase discharge. These actions of epinephrine were associated with a concomitant elevation of cyclic AMP levels. In contrast to the actions of catecholamines and cyclic AMP, acetylcholine and cyclic GMP accelerated lysosomal enzyme release without affecting particle uptake. The actions of acetylcholine were associated with a concomitant elevation of cyclic GMP levels. Increases in neutrophil levels of cyclic GMP but not of cyclic AMP were associated also with the discharge of ß-glucuronidase provoked by particles in the absence of added cholinergic agents. The data suggest that the immunologic release of lysosomal enzymes from human neutrophils can be regulated by autonomic neurohormones, perhaps via the selective formation of appropriate nucleotides.

scholarly journals Regulation of macrophage lysosomal enzyme secretion: role of arachidonate metabolites, divalent cations and cyclic AMP

1980 ◽  
Vol 44 (1) ◽  
pp. 299-315
Author(s):  
R.M. McMillan ◽  
D.E. Macintyre ◽  
J.E. Beesley ◽  
J.L. Gordon
Keyword(s):  
Cyclic Amp ◽  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Divalent Cations ◽  
Cell Types ◽  
Enzyme Secretion ◽  
Ionophore A23187 ◽  
Enzyme Release ◽  
Arachidonate Metabolites

We have investigated the role in macrophage lysosomal enzyme release of arachidonate metabolites, extracellular divalent cations and cyclic AMP (cAMP) which modulate secretion in other cell types. Lysosomal enzyme secretion induced by zymosan was accompanied by release of malondialdehyde (MDA), which is derived from arachidonic acid via prostaglandin synthase. Blockade of MDA formation, by aspirin or indomethacin, was associated with only a small inhibitory effect on lysosomal enzyme release by zymosan: arachidonate metabolites thus play only a minor role in mediating macrophage lysosomal enzyme release. Zymosan-induced secretion of lysosomal enzymes from macrophages did not require extracellular magnesium or calcium although release was enhanced by magnesium and inhibited by calcium. These effects may be related to an influence of the ions on phagocytosis. Elevation of intracellular divalent cation concentrations, by ionophore A23187, induced release of lysosomal enzymes but this was a result of cell lysis. Adenylate cyclase stimulants and dibutyryl cAMP produced slight inhibition of zymosan-induced lysosomal enzyme release. Aminophylline and papaverine caused more marked inhibition but their effects may be due to actions independent of phosphodiesterase inhibition. Our data indicate that arachidonate metabolites and cAMP do not play a major role in regulating zymosan-induced enzyme release from macrophages. Extracellular calcium and magnesium may modulate secretion but the role of intracellular divalent cations remains to be established. We conclude that macrophage lysosomal enzyme secretion is controlled by regulatory mechanisms different from those which control similar degranulation processes in other cell types.

YIN/YANG MODULATION OF LYSOSOMAL ENZYME RELEASE FROM POLYMORPHONUCLEAR LEUKOCYTES BY CYCLIC NUCLEOTIDES

1975 ◽  
Vol 256 (1 Mechanisms of) ◽  
pp. 222-232 ◽  
Author(s):  
Gerald Weissmann ◽  
Ira Goldstein ◽  
Sylvia Hoffstein ◽  
Geneviéve Chauvet ◽  
Roger Robineaux
Keyword(s):  
Lysosomal Enzyme ◽  
Cyclic Nucleotides ◽  
Polymorphonuclear Leukocytes ◽  
Enzyme Release ◽  
Yin Yang
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