muramidase and Granulomatous-Disease--Chronic

Antimicrobial action of polymorphonuclear leukocytes depends on an array of substances carried in their cell membranes and cytoplasmic granules. These substances mediate killing in several systems. Some depend on molecular oxygen while others are independent of it. Some of the systems that depend on oxygen also require myeloperoxidase. The different systems seem able to act in primary or in reserve capacities backing each other up in times of stress or failure. Thus a deficiency in one system does not necessarily leave the polymorphs completely incapable of antimicrobial action although impairment may be severe as in chronic granulomatous disease. In general a poorly functioning polymorph is better than no polymorph at all.

Topics: Animals; Bacteria; Bacterial Physiological Phenomena; Blood Bactericidal Activity; Blood Proteins; Cations; Cytoplasmic Granules; Escherichia coli; Granulocytes; Granulomatous Disease, Chronic; Humans; Hydrogen; Hydrogen Peroxide; Hydrogen-Ion Concentration; Hydroxides; Hydroxyl Radical; Iodine; Lactates; Lactic Acid; Lactoferrin; Leukocytes; Models, Biological; Muramidase; Neutrophils; Opsonin Proteins; Oxygen Consumption; Peptide Hydrolases; Peroxidase; Phagocytosis; Superoxides

Studies were performed to elucidate further the phenomenon of secretagogue-mediated enhancement in the binding of the chemoattractant f-met-leu-[3H]phe to human neutrophils (PMN). Specific f-met-leu-[3H]phe binding to unstimulated PMN reached maximum levels after 10 to 15 min of incubation at 0 degrees C with a saturating concentration of peptide, and consisted of a readily displaceable and a nondisplaceable component. PMN, preexposed to A23187 (2.5 X 10(-8) M) or PMA (0.5 ng/ml) for 30 min at 37 degrees C to stimulate limited and preferential release of specific (secondary) granules (10 to 20% of total lysozyme, no beta-glucuronidase), demonstrated an approximate doubling in the displaceable component of f-met-leu-["3H]phe binding, accompanied by an increasing nondisplaceable component that could not be explained by bulk pinocytosis of extracellular fluid (assessed by [3H]sucrose uptake). The increase in f-met-leu-[3H]phe binding was not affected by inhibitors of protein synthesis, could not be attributed to the secreted products lysosyme or lactoferrin acting on the cell, and, on the basis of studies with PMN from patients with chronic granulomatous disease, could not be attributed to the effects of reactive oxygen species generated in low concentration during stimulation. Functional studies on PMN indicated that preexposure to secretagogues at concentrations demonstrated to increase receptor availability also enhanced subsequent f-met-leu-phe-mediated superoxide and hydrogen peroxide generation. The present data demonstrate that secretagogues may activate PMN to enhance their subsequent responses in f-met-leu-phe-mediated processes, and, combined with previous reports, support the concept that specific granules provide a source of preformed membrane and receptor material that is translocated to the cell surface during the secretion associated with directed locomotion.

Topics: 1-Butanol; Butanols; Calcimycin; Calcium; Chemotaxis, Leukocyte; Cycloheximide; Cytoplasmic Granules; Granulomatous Disease, Chronic; Humans; Hydrogen Peroxide; Lactoferrin; Methionine; Muramidase; N-Formylmethionine; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Oligopeptides; Pinocytosis; Puromycin; Receptors, Immunologic; Superoxides; Tetradecanoylphorbol Acetate

Incubation of human neutrophils with phospholipase C from Clostridium perfringens caused an increase in the ability of the treated cells to bind the chemotactic peptide, F-Met-Leu-Phe. The increase in binding was related to an increase in specific binding of the ligand. The increase in specific binding was, in turn, related to an increased number of peptide receptors. The dissociation constant (KD) for the tripeptide was not altered, on the average, by enzyme treatment. The increase in peptide receptor number was related temporally, and possibly mechanistically, to enzyme-stimulated secretory function involving the secondary granules. Phospholipase C treatment did not similarly augment binding of the complement-derived attractant, C5a. Receptor numbers for different chemotactic ligands may therefore be controlled by different mechanisms. Supplementary experiments provided evidence that this phenomenon was attributable to phospholipase C activity and not to contaminating protease(s).

Topics: Complement C5; Complement C5a; Granulomatous Disease, Chronic; Humans; Kinetics; Muramidase; Neutrophils; Oxygen Consumption; Peptide Hydrolases; Peptides; Peroxidase; Phospholipases; Type C Phospholipases

Topics: Granuloma; Granulomatosis with Polyangiitis; Granulomatous Disease, Chronic; Humans; Lymphomatoid Granulomatosis; Lymphoproliferative Disorders; Muramidase; Peptidyl-Dipeptidase A; Sarcoidosis

During phagocytosis, neutrophils generate reactive oxygen metabolites and release lysosomal enzymes into the extracellular medium. We have investigated the possibility that these enzyme are inactivated by the oxygen compounds. Phagocytosing neutrophils from 12 patients with chronic granulomatous disease, which do not generate these oxygen metabolites, released two to three times more activity of lysozyme and beta-glucuronidase than did normal neutrophils. This difference proved to be due to a decrease of approximately 20% of the total activity of these enzymes in normal neutrophils, but not in neutrophils of patients with chronic granulomatous disease. This inactivation of enzymes took place during phagocytosis of opsonized zymosan particles as well as during stimulation of normal cells with phorbol myristate acetate. The inactivation was not due to formation of inhibitors. The lysosomal enzymes were not activated when the neutrophils were stimulated under anaerobic conditions. Addition of catalase, superoxide dismutase, or albumin gave no protection against the oxidative damage; reduced glutathione gave partial protection. The oxidative inactivation was more pronounced in the presence of azide. Measurement of the activity and the amount of protein of acid alpha-glucosidase in the cells showed that the specific activity of this enzyme decreased by approximately 50% during 30 min of phagocytosis. This indicates that the inactivation of the lysosomal enzymes takes place in the phagolysosomes, before the enzymes have leaked into the extracellular medium.

Topics: Extracellular Space; Glucuronidase; Glutathione; Granulomatous Disease, Chronic; Humans; L-Lactate Dehydrogenase; Lysosomes; Muramidase; Neutrophils; Oxidation-Reduction; Phagocytosis

Topics: Adenosine Triphosphate; Bacterial Proteins; Blood Bactericidal Activity; Chediak-Higashi Syndrome; Child; Glucose; Granulomatous Disease, Chronic; Halogens; Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; Microscopy, Electron; Muramidase; Neutrophils; Oxygen Consumption; Peroxidases; Phagocytosis; Staphylococcus; Vacuoles