cyclic-gmp and Chediak-Higashi-Syndrome

Like other cells, leukocytes have the cellular skeleton comprising microtubules, actine, myosine, and intermediate filaments. Elements of the cellular skeleton are connected with receptors on the plasma membrane surface and determine the distribution density of these receptors. There is functional relationship between microtubules and actine filaments, in particular, microtubules determine uniform distribution of actine filaments in the cell cytoplasm. The basic principle of functioning of the cellular skeleton consists of the process of polymerization and reversible depolymerization of proteins forming components of the cellular skeleton. These processes are regulated by Ca2+, calmoduline, as well as by the ratio of cyclic adenosine monophosphate and guanidine monophosphate in the cell. The cellular skeleton determines the most important functions of leukocytes: their mobility, binding and absorption of various substances, processes of degranulation, fusion of granules with phagocytic vacuole. Cellular skeleton defects are accompanied by recurrent bacterial infections. Several such defects are known: leukocyte actine dysfunction syndrome, Chediak-Higashi syndrome, syndrome with marked increase in the content of cGMP and microtubules in leukocytes. In these syndromes, the therapeutic effect is achieved with the substances which regulate the level of cyclic nucleotides in leukocytes, among them large doses of ascorbic acid.

Topics: Arachidonic Acids; Cell Membrane; Chediak-Higashi Syndrome; Cyclic AMP; Cyclic GMP; Cytoskeleton; Humans; Leukocytes; Microtubules; Phagocytosis

Topics: Animals; Ascorbic Acid; Chediak-Higashi Syndrome; Concanavalin A; Cyclic GMP; Disease Models, Animal; Humans; Immunity, Cellular; Leukocytes; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Prostaglandins

Intensive laboratory investigation of patients with recurrent infections, and with infections with microbial species not usually considered to be pathogenic, have led to the identification of several defects in granulocyte function. The two functions of granulocytes which have received most attention in the past decade have been locomotion (especially response to chemotactic stimulation) and microbicidal activity. Defective granulocyte chemotaxis has been demonstrated in patients with clinical manifestations suggesting abnormalities related to vasoactive amines, i.e., patients with eczema and extreme IgE hyperimmunoglobulinemia. The depressed granulocyte chemotactic responsiveness found in these patients can be reproduced in vitro when histamine and beta adrenergic agents are incubated with control granulocytes. Since these compounds have been shown to increase levels of intracellular cyclic AMP in other cells, there appears to be an association between cyclic nucleotide metabolism and regulation of granulocyte locomotion. Defective granulocyte microbicidal activity is found in patients with chronic granulomatous disease and it has been shown that there is little increase in oxidative metabolism during phagocytosis by these cells. Methods for quantitating the oxidative metabolism of granulocytes and monocytes include oxygen uptake, reduction of nitroblue tetrazolium, formate oxidation, and chemiluminescence response during phagocytosis. Since products of oxygen metabolism, i.e., hydrogen peroxide, superoxide or singlet oxygen do not accumulate in granulocyte phagocytic vacuoles, intracellular microbes are not killed (except bacterial species that produce hydrogen peroxide). The biochemical basis for defective oxidative metabolism in granulocytes from patients with chronic granulomatous disease appears to be associated with abnormal nucleotide oxidase activity.

Topics: Blood Bactericidal Activity; Carbohydrate Metabolism, Inborn Errors; Cell Movement; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Cyclic AMP; Cyclic GMP; Granulocytes; Granulomatous Disease, Chronic; Humans; Hydrogen Peroxide; Hydrogen-Ion Concentration; Hydrolases; Leukocytes; Luminescent Measurements; Mannose; Metabolism, Inborn Errors; NADH, NADPH Oxidoreductases; NADP; NADPH Oxidases; Nucleotides, Cyclic; Oxygen Consumption; Peroxidase; Phagocyte Bactericidal Dysfunction; Phagocytosis; Protein-Energy Malnutrition

Topics: Ascorbic Acid; Blood Bactericidal Activity; Chediak-Higashi Syndrome; Chemotaxis; Cyclic AMP; Cyclic GMP; Humans; Infant; Leukocyte Count; Leukocytes

All leukocytes are capable of responding chemotactically (oriented locomotion) and chemokinetically (stimulated nondirected or random locomotion) to a variety of chemical agents. A brief review of the in vitro and in vivo methods of studying neutrophil movement and our present knowledge of chemotactic factors is presented as well as a discussion on the mechanisms of stimulated movement. Two clinically important instances of defects in neutrophil movement, ie, the Chédiak-Higashi syndrome and a case of actin dysfunction, are herein described.

Topics: Ascorbic Acid; Calcium; Cell Movement; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Complement C3; Complement C5; Cyclic AMP; Cyclic GMP; Cytological Techniques; Cytoskeleton; Humans; Inflammation; Kallikreins; Lysosomes; Microtubules; Neutrophils; Oligopeptides; Plasminogen Activators; Receptors, Drug; Skin Window Technique; Sodium; Structure-Activity Relationship

The Chediak-Higashi (CH) syndrome of man and several animal species is characterized by the presence of abnormal giant granules in all granule-containing cells and by defects in chemotaxis and lysosomal degranulation during phagocytosis in polymorphonuclear leukocytes (PMNs). Since similar functional abnormalities have been reported in normal PMNs following exposure to colchicine and other agents that disrupt microtubles it was proposed that microtubule function may be impaired in the CH syndrome. The mobility of concanavalin A (con A)-receptor complexes on PMN membranes was used to test microtubule integrity. Normal PMNs showed a uniform distribution of membrane-bound con A. By contrast, con A was aggregated into surface caps on both colchicine-treated normal PMNs and untreated PMNs from mice and a patient with CH syndrome. This result is consistent with impaired microtubule function in the CH cells. The spontaneous capping response of CH PMNs was inhibited by cyclic GMP and by cholinergic agonists that can elevate cyclic GMP levels in neutrophils. This raised the possibility that the microtubule defect in CH cells may be correctable by treatments that increase cyclic GMP generation. Direct evidence for both the absence of microtubule assembly in con A-treated PMNs from the CH patient and for normal microtubule assembly in CH PMNs incubated with cyclic GMP and cholinergic agonists prior to con A treatment was obtained by electron microscopy. In addition, evidence for a direct relationship between the microtubule defect and the development of giant lysosomes in CH cells was obtained. Thus, CH fibroblasts grown in vitro developed abnormal lysosomes in the majority of cells. However, the same cells cultured in the presence of cholinergic agonists developed a majority of lysosomes that were morphologically normal at the level of the light microscope. Similarly, granule morphology appeared normal in peripheral blood leukocytes from mice treated chronically in vivo with cholinergic agonists.

Topics: Animals; Chediak-Higashi Syndrome; Chemotaxis; Colchicine; Concanavalin A; Cyclic AMP; Cyclic GMP; Cytoplasmic Granules; Glycoproteins; Humans; Mice; Microtubules; Parasympatholytics; Phagocytosis; Receptors, Drug

Topics: Adult; Atropine; Bucladesine; Carbachol; Cell Line; Cell Survival; Chediak-Higashi Syndrome; Cyclic GMP; Cytotoxicity, Immunologic; Dibutyryl Cyclic GMP; Humans; Immunity, Cellular; Incubators; Kinetics; Lymphocytes; Male

Topics: Adult; Arthritis, Rheumatoid; Ascorbic Acid; B-Lymphocytes; Chediak-Higashi Syndrome; Concanavalin A; Cyclic GMP; Humans; Hypersensitivity, Delayed; Immunity, Cellular; Immunoglobulins; In Vitro Techniques; Lymphocytes; Middle Aged; Monocytes; Pokeweed Mitogens; T-Lymphocytes, Regulatory

A diminished chemotactic response was observed with the neutrophils of a patient with the Chediak-Higashi syndrome, who was not in the accelerated phase of the disease. An abnormally low release of myeloperoxidase from these cells during phagocytosis was also noted; this resulted in a decreased iodination capacity and probably also caused the defect in the intracellular killing of bacteria by the neutrophils. The level of cyclic AMP in these cells was elevated, but decreased after treatment with ascorbate either in vitro or in vivo. During ascorbate therapy, the bactericidal activity of the neutrophils normalized, whereas the chemotactic response remained low. Nevertheless, the patient had significantly less infections during ascorbate therapy. The bleeding tendency, due to a storage-pool disorder of the Chediak-Higashi platelets, was unaffected by treatment with ascorbate. The patient's lymphocytes did not display any activity in antibody-dependent lymphocytotoxicity. This defect was not affected by treatment with ascorbate either.

Topics: Adenosine Diphosphate; Adolescent; Ascorbic Acid; Bleeding Time; Blood Platelets; Cell Movement; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Cyclic AMP; Cyclic GMP; Humans; Hypersensitivity, Delayed; Lymphocytes; Male; Neutrophils; Nucleotides, Cyclic; Peroxidase

Topics: Animals; Arachidonic Acids; Binding Sites; Calcium; Cattle; Chediak-Higashi Syndrome; Chemotactic Factors; Chemotaxis; Chemotaxis, Leukocyte; Complement C5; Cyclic AMP; Cyclic GMP; Cytochalasin B; Humans; Hypergammaglobulinemia; Kartagener Syndrome; Mice; Oligopeptides; Potassium; Rabbits; Syndrome

Topics: Ascorbic Acid; Chediak-Higashi Syndrome; Cyclic AMP; Cyclic GMP; Humans; Lymphocytes; Monocytes; Neutrophils

Topics: Adult; Animals; Ascorbic Acid; Blood Bactericidal Activity; Candidiasis; Cell Movement; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Cyclic AMP; Cyclic GMP; Humans; Leukocytes; Lymphocyte Activation; Male; Mice; Mice, Inbred C57BL; Neutrophils

Topics: Chediak-Higashi Syndrome; Child, Preschool; Cyclic GMP; Female; Humans; Lymphocytes; Lysosomes; Microscopy, Electron; Microtubules; Neutrophils

The beige mouse, C57BL/6 (bg/bg), is an animal model for the Chediak-Higashi syndrome in man, a disease characterized morphologically by giant lysosomes in most cell types. Half-lives for the turnover of [(14)C]bicarbonate-labeled total soluble liver protein were determined in normal and beige mice. No significant differences were observed between the normal and mutant strain for both rapidly and slowly turning-over classes of proteins. Glucagon treatment during the time-course of protein degradation had similar effects on both normal and mutant strains and led to the conclusion that the rate of turnover of endogenous intracellular protein in the beige mouse liver does not differ from normal. The rates of uptake and degradation of an exogenous protein were determined in normal and beige mice by intravenously injecting (125)I-bovine serum albumin and following, in peripheral blood, the loss with time of phosphotungstic acid-insoluble bovine serum albumin and the parallel appearance of phosphotungstic acid-soluble (degraded) material. No significant differences were observed between beige and normal mice in the uptake by liver lysosomes of (125)I-bovine serum albumin (t((1/2)) = 3.9 and 2.8 h, respectively). However, it was found that lysosomes from livers of beige mice released phosphotungstic acid-soluble radioactivity at a rate significantly slower than normal (t((1/2)) = 6.8 and 3.1 h, respectively). This defect in beige mice could be corrected by chronic administration of carbamyl choline (t((1/2)) = 3.5 h), a cholinergic agonist which raises intracellular cyclic GMP levels. However, no significant differences between normal and beige mice were observed either in the ability of soluble extracts of liver and kidney to bind [(3)H]cyclic GMP in vitro or in the basal levels of cyclic AMP in both tissues. The relevance of these observations to the presumed biochemical defect underlying the Chediak-Higashi syndrome is discussed.

Topics: Animals; Chediak-Higashi Syndrome; Cyclic AMP; Cyclic GMP; Half-Life; Liver; Lysosomes; Male; Mice; Mice, Inbred C57BL; Protein Binding; Proteins; Serum Albumin, Bovine

The adherence of human polymorphonuclear leukocytes (PMN) to nylon fibers is inhibited in a dose-dependent fashion by exposure in vitro of these cells to either colchicine or VM-26, both of which agents prevent microtubule assembly. Mean adherence of human PMN was 48% +/- 2%, following treatment with 10(-5) M colchicine or 10(-4) M VM-26 it was reduced to 31% +/- 2% and 7%, respectively. Peritoneal PMN obtained from mice and mink with Chediak-Higashi syndrome (CHS) thought to have a microtubule-membrane disorder affecting the PMN had a mean adherence of 29% +/- 3% and 40% +/- 8% compared to control values of 46% +/- 5% and 73% +/- 8%, respectively, from the mice and mink. Both ascorbic acid and bethanechol, shown previously to enhance microtubule assembly in humans with CHS, normalized granulocyte adherence in PMN obtained from mice with CHS. Cyclic nucleotide levels were not altered by treatment of human PMN with colchicine, nor did they differ between normal and CHS animals. Thus it appears that the state of microtubule assembly may directly affect the properties of the PMN plasma membrane without requiring alterations of cyclic nucleotides as an intermediary.

Topics: Animals; Bethanechol Compounds; Cell Adhesion; Chediak-Higashi Syndrome; Colchicine; Cyclic AMP; Cyclic GMP; Dose-Response Relationship, Drug; Female; Granulocytes; Humans; Leukocytes; Male; Mice; Mice, Inbred C57BL; Microtubules; Mink; Neutrophils

The addition of cholinergic agents and cyclic 3'5'-guanosine monophosphate (cGMP) to polymorphonuclear leukocytes in vitro from a patient with Chediak-Higashi syndrome corrected the impaired release of the lysosomal enzyme, beta-glucuronidase, to normal. Coinciding with the improvement in degranulation, the bactericidal capacity was enhanced to normal. Similar concentrations of cholinergic agents potentiated chemotaxis to control values. On the other hand, the phagocytic rate of lipopolysaccharide-coated paraffin-oil droplets was not altered by the cholinergic agents. The improvement in Chediak-Higashi syndrome polymorphonuclear leukocyte function by the addition of cholinergic agents and dibutyryl cGMP suggested disturbed intracellular cyclic nucleotide levels.

Topics: Blood Bactericidal Activity; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Cyclic GMP; Female; Glucuronidase; Humans; Infant; Leukocytes; Lysosomes

The anti-helminthic drug levamisole hydrochloride has been reported to stimulate immune responses in humans and experimental animals. We have investigated levamisole effects on human leukocyte locomotion in vitro in studies of neutrophils and mononuclear cells from normal adults, from patients with Chediak-Higashi disease and from patients with the syndrome of hyperimmunoglobulin E, recurrent pyogenic infections, and defective leukocyte chemotaxis. Directed migration (chemotaxis) of neutrophils and mononuclear cells from normal adults and from the hyperimmunoglobulin E syndrome patients, but not from Chediak-Higashi patients, were stimulated by levamisole at concentrations of 0.01-1.0 micronM, with stimulation observed most consistently at 0.1 micronM. These concentrations of drug also increased cyclic GMP levels in mononuclear cells and enhanced hexose monophosphate shunt activity in neutrophils, but did not alter chemotactic factor-induced changes in the surface charge of neutrophils. Other concentrations of levamisole did not affect leukocyte locomotion except for a high concentration (5.0 mM) which stimulated both random and directed leukocyte migration. When patients with the hyperimmunoglobulin E syndrome took levamisole by mouth, the abnormal chemotactic responses of their neutrophils were significantly improved towards normal. These studies are the first to show pharmacologic improvement of in vitro leukocyte locomotion in patients in whom recurrent infections have been attributed to a defect of this leukocyte function.

Topics: Adolescent; Adult; Chediak-Higashi Syndrome; Chemotaxis, Leukocyte; Child; Cyclic AMP; Cyclic GMP; Female; Glucose; Humans; Hypergammaglobulinemia; Immunoglobulin E; Leukocytes; Levamisole; Male

Because ascorbate potentiates chemotaxis of normal leukocytes, we examined the effect of ascorbate on polymorphonuclear leukocytes from a patient with the Chediak-Higashi syndrome. Chemotactic migration was 104+/-16 leukocytes per 10 fields (mean+/-S.D.) initially and 258+/-44 (P less than 0.001) after ascorbate, as compared to 182+/-10 in controls. There was no bactericidal activity by 40 minutes in the patient's untreated leukocytes. After ascorbate bactericidal activity of patient and untreated control cells was the same. The addition of ascorbate reduced cAMP levels in the patient's cells from a mean of 34.5 pmoles per 10(7) polymorphonuclear leukocytes to 5.9, as compared to a control value of 3.1+/-1.4. The association of elevated cAMP and impaired function in the polymorphonuclear leukocytes of patients with the Chediak-Higashi syndrome may be related to abnormal microtubular assembly.

Topics: Ascorbic Acid; Blood Bactericidal Activity; Chediak-Higashi Syndrome; Cyclic AMP; Cyclic GMP; Female; Glucuronidase; Humans; In Vitro Techniques; Infant; Leukocytes; Microtubules; Monocytes; Stimulation, Chemical

Topics: Animals; Binding Sites, Antibody; Cell Membrane; Chediak-Higashi Syndrome; Colchicine; Concanavalin A; Cyclic GMP; Disease Models, Animal; Leukocytes; Mice