muramidase and Chediak-Higashi-Syndrome

Topics: Acid Phosphatase; Agranulocytosis; Blood Bactericidal Activity; Chediak-Higashi Syndrome; Chemotaxis; Child, Preschool; Complement System Proteins; Cytoplasmic Granules; Female; Glucosephosphate Dehydrogenase Deficiency; Humans; Immunologic Deficiency Syndromes; Infant, Newborn; Infant, Premature; Leukocytes; Liver; Lysosomes; Macrophages; Male; Monocytes; Mononuclear Phagocyte System; Muramidase; NADH, NADPH Oxidoreductases; Neutrophils; Opsonin Proteins; Peroxidases; Phagocyte Bactericidal Dysfunction; Phagocytosis; Spleen

Topics: Bacterial Infections; Burns; Chediak-Higashi Syndrome; D-Amino-Acid Oxidase; Glutathione; Glutathione Reductase; Hexoses; Humans; Infant, Newborn; Leukocytes; Lipids; Muramidase; NAD; NADP; Neutrophils; Oxidation-Reduction; Peroxidases; Phagocyte Bactericidal Dysfunction; Proteins

The abnormal giant granules of Chediak-Higashi syndrome (CHS) neutrophils in humans are thought to be derived from both azurophil and specific granules, whereas the presence of gelatinase granules and their contribution to giant granule formation has not been investigated previously. We have examined the ultrastructure and mobilization of neutrophil granules from a patient with CHS by immunogold electron microscopy and exocytosis experiments of isolated leukocytes. The giant granules contained the azurophil granule components myeloperoxidase and CD63. We found no evidence of involvement of specific or gelatinase granules in the formation of giant granules because lactoferrin and gelatinase were contained in normal-appearing peroxidase-negative granules. On stimulation of leukocytes with N-formyl-methionyl-leucyl-phenylalanine and the calcium ionophore, ionomycin, there was a diminished exocytosis of myeloperoxidase in CHS compared with a healthy control, indicating a lack of mobilization of the giant granules. On the other hand, there was a normal or augmented release of lactoferrin and gelatinase in CHS neutrophils, with gelatinase granules being the most easily mobilized, as known from normal neutrophils. In conclusion, giant granules from CHS neutrophils originate from azurophil granules but not from the specific and gelatinase granules.

Topics: Antibody Specificity; Azure Stains; Chediak-Higashi Syndrome; Child, Preschool; Cytoplasmic Granules; Exocytosis; Female; Gelatinases; Humans; Immunohistochemistry; Microscopy, Electron; Muramidase; Neutrophils; Peroxidase

Neutrophils from two Chédiak-Higashi syndrome brothers were isolated, suspended in heparinized sucrose, lysed, and filtered. The granule-rich filtrate was centrifuged on a sucrose gradient (rho = 1.287-1.10 g/ml) at a mean force of 95,000g for 4 hours. The gradients contained one band at rho = 1.18 g/ml (band C) which was broader than normal and lacked normal bands A, rho = 1.22 g/ml, and B, rho = 1.20 g/ml. Gradient fractions were assayed for enzyme activities and protein. No marker enzymes identified densities normally occupied by bands A and B, and of the enzymes measured, only lysozyme showed peak activity with band C. Thus, only normal specific granules were present. Two azurophil granules, normally present and separable, were absent. Also identified was eosinophil granule peroxidase at rho = 1.24 g/ml (band E). Alkaline phosphatase, not a granule marker, was twice normal at the normal density, rho = 1.14-1.15 g/ml, consistent with an increase in unidentified membranes. A lysate gradient suggested that the giant azurophilic granules were rho = 1.25-1.27 g/ml. These neutrophils contain blue-grey or slate-grey giant granules, which are not truly azurophilic or basophilic, but should continue to be identified as azurophilic to conform to the convention making "azurophilic" and "peroxidase-positive" synonymous. The eosinophils contain normal eosinophil granules as well as giant inclusion granules. In contrast, neutrophils are deficient in both normal azurophilic granules.

Topics: Adult; Alkaline Phosphatase; Cell Fractionation; Cell Separation; Centrifugation, Density Gradient; Chediak-Higashi Syndrome; Cytoplasmic Granules; Eosinophils; Humans; Isoenzymes; Male; Muramidase; Neutrophils; Peroxidase; Peroxidases; Staining and Labeling

The proximal nephron of C57 beige mice with a genetic defect analagous to the Chédiak-Higashi syndrome (CHS) has been compared with that of normal C57 black mice. The concanavalin A-horseradish peroxidase (Con A-HRP) technique stained the brush border of the proximal straight tubule heavily in black mice and weakly in beige mice. In beige mice this method stained the brush border of the proximal convoluted tubules weakly and the brush border of the proximal straight tubules only negligibly. Periodic acid-Schiff staining showed no such difference between beige and black mice but revealed an increase distally in the size of the CHS inclusions in the proximal straight tubule of beige mice. Immunostaining visualized abundant lysozyme in the first portion of the proximal nephron but none in the more distal segments of beige and black mice alike. At the ultrastructural level, the proximal convoluted tubules of black mice contained two morphologic types of heterophagosomes, which apparently differed in accord with the stage of their development. Proximal straight tubules contained morphologically different heterophagic bodies. The mature stages of these heterophagosomes were greatly enlarged in CHS mice. With the periodic acid-thiocarbohydrazide-silver proteinate (PA-T-SP) method for localizing glycoprotein ultrastructurally, the microvillar brush border, apical invaginations of the plasmalemma, Golgi cisternae, and lysosomal inclusions stained selectively in the proximal nephron in both strains. The proximal straight nephron of beige mice after staining with the PA-T-SP method appeared depleted of the strongly reactive apical invaginations in some areas, particularly where large heterophagosomes bordered the apical plasmalemma. The enlarged secondary lysosomes of heterophagic origin in beige mice varied in showing both diffuse and focal PA-T-SP reactivity. Lysosomal acid phosphatase activity appeared decreased, and peroxisomes were normal in size but increased in number in the proximal nephron of beige mice.

Topics: Animals; Chediak-Higashi Syndrome; Disease Models, Animal; Immunoenzyme Techniques; Inclusion Bodies; Kidney Tubules, Proximal; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Microscopy, Electron; Muramidase; Phagocytes

The tissue damage induced by murine cytomegalovirus (MCMV) in mice with the beige mutation (bg/bg) and in their normal littermates (bg/ +) was investigated. The beige mutation in mice is a homolog of the Chédiak-Higashi syndrome in man, and various dysfunctions of phagocytes and decreased activity of natural killer cells have been demonstrated in these animals. Tissue damage, especially in the liver and spleen, was more conspicuous in bg/bg than in bg/ + mice and was associated with frequent intranuclear inclusions and a higher titer of virus. However, the mutation did not appear to alter the organ distribution of tissue damage induced by MCMV. The inflammatory response in the liver, which is presumed to contribute to host resistance, appeared under certain circumstances to be delayed and deficient in bg/bg mice.

Topics: Animals; Aspartate Aminotransferases; Chediak-Higashi Syndrome; Cytomegalovirus Infections; Disease Models, Animal; Disease Susceptibility; Female; Hematologic Tests; Kidney; Liver; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Muramidase; Mutation; Myocardium; Spleen; Thymus Gland

Topics: Animals; Carbohydrates; Cell Membrane; Chediak-Higashi Syndrome; Histocytochemistry; Mice; Microbodies; Muramidase; Organoids; Pulmonary Alveoli

A method of blood granylocyte concentration and isolation of granules from both normal and neutropenic Chediak-Higashi syndrome (CHS) patients is described. the intracellular distribution of activity for several hydrolases in CHS granulocytes differs from normal; significantly more activity is present in the cytoplasmic fraction and correspondingly less is granule-associated. Isolated CHS granules are not more sensitive to the labilizing agents vitamin A, progesterone, or etiocholanolone. Specific activities of myeloperoxidase and ss-glucuronidase in CHS granulocytes are lower than normal while alkaline phosphatase is elevated. Other lysosomal enzyme activities are normal. Lysosomal enzyme distribution and content are similar in CHS and normal mononuclear cells. The possible significance of these findings is discussed.

Topics: Acid Phosphatase; Alkaline Phosphatase; Cathepsins; Chediak-Higashi Syndrome; Cytoplasmic Granules; Etiocholanolone; Female; Glucuronidase; Heparin; Humans; Leukocytes; Lysosomes; Male; Muramidase; Peroxidase; Progesterone; Vitamin A

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

Topics: Acid Phosphatase; Adult; Agranulocytosis; Alkaline Phosphatase; Bacterial Infections; Chediak-Higashi Syndrome; Chemotaxis; Female; Glucuronidase; Humans; Leukocytes; Lysosomes; Male; Muramidase; Peroxidases; Skin Window Technique; Staphylococcal Infections; Streptococcal Infections