Abstract
The disruption of the molecular organization of the plasma membrane of leukocytes by phagocytosable particles, or by agents such as surfactants, antibodies, phospholipase C, fatty acids and chemotactic factors, leads to a stimulation of the phagocyte oxidative metabolism. Concanavalin A (Con A) has been used as a tool to study the mechanism of this metabolic regulation.
The binding of Con A to the surface of polymorphonuclear leukocytes (PMNL) or macrophages produces a rapid enhancement of oxygen uptake and glucose oxidation through the hexose monophosphate pathway (HMP). This is explained by an activation of the granular NADPH oxidase, the key enzyme in the metabolic stimulation. The effect of Con A is not due to endocytosed lectin, since Con A covalently coupled to large sepharose beads still acts as stimulant.
The metabolic changes caused by Con A are reversible. If, after the onset of stimulation, sugars with high affinity for Con A are added to the leukocyte suspension, the activity of granular NADPH oxidase and the rate of respiration and glucose oxidation return to their resting values.
The metabolic burst, while partially supressed by treatment of PMNL with iodoacetate, sodium fluoride and cytochalasin B, is slightly increased by colchicine.
Con A induces a selective release of granular enzymes (β-glucuronidase, peroxidase, alkaline phosphatase) from PMNL, whereas no leakage of cytoplasmic enzymes is observed. The enzyme release is inhibited by iodoacetate and by drugs known to increase cell levels of cyclic AMP.
Based on a current view of the mode of interaction between Con A and cell surfaces, a model of the metabolic disruption of leukocytes is presented.
This work was aided by a grant from the Italian National Research Council (No. 73.00441.04).
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References
Aaronson, S. A., and Todaro, G. J. (1968). “Basis for the acquisition of malignant potential by mouse cells cultivated in vitro.” Science 162, 1024.
Azzi, A. (1969). “Redistribution of the electrical charge of the mitochondrial membrane during energy conservation.” Biochem. Biophys. Res. Commun. 37, 254.
Bergmayer, H. U., Bernt, E., and Hesso, B. (1965). “Lactic dehydrogenase.” In Methods of enzymatic analysis ( H. U. Bergmayer, ed.), Academic Press, New York.
Berlin, R. D. (1972). “Effect of Concanavalin A on phagocytosis.” Nature New Biology 235, 44.
Berlin, R. D., Oliver, J. M., Ukena, T. E., and Yin, H. H. (1974). “Control of cell surface topography.” Nature New Biology 247, 45.
Burger, M. M. (1970). “A difference in the architecture of the surface membrane of normal and virally transformed cells.” Proc. Nat. Acad. Sci. U. S. A. 62, 994.
Cooper, R. H., Ashcroft, S. J., and Randle, P. J. (1973). “Concentrations of adenosine 3’:5’ - cyclic monophosphate in mouse pancreatic islets measured by a protein-binding radioassay.” Biochem. J. 134, 599.
Cuatrecasas, P. (1969). “Interaction of insulin with the cell membrane: the primary action of insulin.” Proc. Nat. Acad. Sci. U. S. A. 63, 450.
Curtis, A. S. G. (1967). The cell surface: its molecular role in morphogenesis. Pergamon Press, Oxford.
Fisch, H. U., Pliska, U., and Schwyzer, R. (1972). “A covalent protein-Sepharose complex for the specific adsorption and assay of adenosine 3’:5’ - monophosphate.” Eur. J. Biochem. 30, 1.
Fortes, P. A. G., and Hoffman, J. F. (1971). “Interactions of the fluorescent anion 1-anilino-8-naphthalene sulfonate with membrane charges in human red cell ghosts.” J. Membrane Biol. 5, 154.
Gianetto, R., and de Duve, C. (1955). “Comparative study of the binding of acid phosphatase, ß-glucuronidase and cathespin by rat-liver particles.” Biochem. J. 59, 433.
Gingell, D. (1973). “Membrane permeability change by aggregation of mobile glycoprotein units.” J. Theor. BioZ. 38, 677.
Goldstein, I. J. (1975). Personal communication.
Goldstein, I. J., Hollerman, C. E., and Smith, E. E. (1965). “Protein-carbohydrate interaction. II. Inhibition studies on the interactions of Concanavalin A with polysaccharides.” Biochemistry 4, 876.
Gomperts, B., Lanteline, F., and Stock, R. (1970). “Ion association reactions with biological membranes studied with the fluorescent dye 1-anilino-8-naphthalene sulfonate.” J. Membrane Biol. 3, 241.
Greaves, M. F., and Bauminger, S. (1972). “Activation of T and B cells by insoluble phytomitogens.” Nature New Biol. 235, 67.
Hecht, Y. P., Dellacha, J. M., Santome, J. A., Paladini, A. C., Hurwitz, E., and Sela, M. (1972). Personal Communication.
Henson, P. M. (1972). “Pathologic mechanisms in neutrophilmediated injury.” Am. J. PathoZ. 68, 593.
Holmes, B., Sater, J., Rodey, G., Park, B., and Good, R. (1969). “Changes in intracellular distribution of lysosomal enzymes during phagocytosis by human polymorphonuclear leukocytes.” Clin. Invest. 48, 39a, 125 abs.
Kornberg, A., and Horecker, B. L. (1955). “Glucose-6-phosphate dehydrogenase.” In Methods in enzymology (S. P. Colowick and N. O. Kaplan, eds.) vol. I.
Ling, N. R. (1968). Lymphocyte stimulation. North Holland Publishing Co., Amsterdam.
Michell, R. H., Karnovsky, M. J., and Karnowsky, M. L. (1970). “The distribution of some granule-associated enzymes in guinea-pig polymorphonuclear leukocytes.” Biochem. J. 116, 207.
Nazakawa, T., Asani, K., Shoger, R., Fujiwara, A., and Yasumasu, I. (1970). “Ca++ uptake, H+ ejection and respiration in sea-urchin eggs on fertilization.” Exp. CeZZ Res. 63, 143.
Nicolson, G. L. (1973). “Anionic sites of human erythrocyte membranes. I. Effects of trypsin, phospholipase C, and pH on the topography of bound positively charged colloidal particles.” J. Cell Biol. 57, 373.
Oliver, J. M., Ukena, T. E., and Berlin, R. D. (1974). “Effects of phagocytosis and colchicine on the distribution of lectinbinding sites on cell surfaces.” Proc. Nat. Acad. Sci. U. S. A. 71, 394.
Pastan, I., Roth, J., and Macchia, V. (1966). “Binding of hormone to tissue: the first step in polypeptide hormone action.” Proc. Nat. Acad. Sci. U. S. A. 56, 1802.
Patriarca, P., Cramer, R., Moncalvo, S., Rossi, F., and Romeo, D. (1971). “Enzymatic basis of metabolic stimulation in leukocytes during phagocytosis: the role of activated NADPH oxidase.” Arch. Biochem. Biophys. 145, 255.
Patriarca, P., Cramer, R., Dri, P., Fant, L., Basford, R. E., and Rossi, F. (1973). “NADPH oxidizing activity in rabbit polymorphonuclear leukocytes: localization in azurophilic granules.” Biochem. Biophys. Res. Commun. 53, 830.
Porath, J., Axen, R., and Ernback, S. (1967). “Chemical coupling of proteins to Agarose.” Nature New Biology 215, 1491.
Rinderknecht, H. (1962). “Ultra-rapid fluorescent labelling of proteins.” Nature New Biology 193, 167.
Romeo, D. (1974). “Modulation of phagocyte metabolism by perturbation of their surface with Concanavalin A.” In Comparative biochemistry and physiology of transport (K. Bloch, L. Bolis, and S. E. Luria, eds.), in press.
Romeo, D., Cramer, R., and Rossi, F. (1970). “Use of 1-anilino8-naphthalene sulfonate to study structural transitions in cell membrane of PMN leukocytes.” Biochem. Biophys. Res. Commun. 41, 582.
Romeo, D., Zabucchi, G., and Rossi, F. (1973a). “Reversible metabolic stimulation of polymorphonuclear leukocytes and macrophages by Concanavalin A.” Nature New Biology 243, 111.
Romeo, D., Zabucchi, G., Marzi, T., and Rossi, F. (1973a). “Kinetic and enzymatic features of metabolic stimulation of alveolar and peritoneal macrophages challenged with bacteria.” Exp. Cell Res. 78, 423.
Romeo, D., Cramer, R., Marzi, T., Soranzo, M. R., Zabucchi, G., and Rossi, F. (1973). “Peroxidase activity of alveolar and peritoneal macrophages.” J. ReticuZoend. Soc. 13, 399.
Romeo, D., Zabucchi, G., Jug, M., and Rossi, F. (1974a). “Alteration of macrophage surface in the course of immunological activation: decay of metabolic response to Concanavalin A.” Cell. Immunol.,in press.
Romeo, D., Jug, J., Zabucchi, G., and Rossi, F. (1974b). “Perturbation of leukocytes metabolism by nonphagocytosable Concanavalin A-coupled beads.” FEBS Letters,in press.
Rossi, F., and Zatti, M. (1964). “Changes in the metabolic pattern of polymorphonuclear leukocytes during phagocytosis.” Brit. J. Exp. PathoZ. 45, 548.
Rossi, F., and Zatti, M. (1968). “Mechanism of the respiratory stimulation in saponine treated leukocytes. The KCN insensitive oxidation of NADPH.” Biochim. Biophys. Acta 153, 296.
Rossi, F., Romeo, D., and Patriarca, P. (1972). “Mechanism of phagocytosis associated oxidative metabolism in polymorphonuclear leukocytes and macrophages.” RES: J. Reticuloend. Soc. 12, 127.
Rossi, F., Patriarca, P., and Romeo, D. (1974). “Regulation of oxidative metabolism and functions of phagocytes.” In Future trends in inflammation (D. A. Willoughby, ed.)
Sbarra, A. J., and Karnovsky, M. L. (1959). “The biochemical basis of phagocytosis.” J. BioZ. Chem. 237, 1355.
Selvaraj, R. J., and Sbarra, A. J. (1966). “Relationship of glycolytic and oxidative metabolism to particle entry and destruction in phagocytosing cells.” Nature New Biology 211, 1272.
Selvaraj, R. J., McRipley, R. J., and Sbarra, A. J. (1967). “The metabolic activities of leukocytes from limphoproliferative and myeloproliferative disorders during phagocytosis.” Cancer Res. 27, 2287.
Simberkoff, M. S., and Elsback, P. (1971). “The interaction in vitro between polymorphonuclear leukocytes and mycoplasma.” J. Exp. Med. 134, 1417.
Singer, S. J., and Nicolson, G. L. (1972). “The fluid mosaic model of the structure of cell membranes.” Science 175, 720.
Smith, C. W. and Hollers, J. C. (1970). “The pattern of binding of fluorescein-labeled Concanavalin A to motil lymphocyte.” J. ReticuZoend. Soc. 8, 458.
Smith, S. B., and Revel, J. (1972). “Mapping of Concanavalin A binding sites on the surface of several cell types.” Develop. BioZ. 27, 434.
Taylor, R. B., Duffus, P. H., Raff, M. C., and de Petris, S. (1971). “Redistribution and pinocytosis of lymphocyte surface immunoglobulins molecules induced by anti-immunoglobulin antibody.” Nature New Biology 233, 225.
Wright, D. G., and Malawista, S. E. (1972). “The mobilization and extracellular release of granular enzymes from human leukocytes during phagocytosis.” J. CeZZ BioZ. 53, 788.
Zigmond, S. H., and Hirsch, J. G. (1972). “Cytochalasin B: inhibition of D-2-deoxyglucose transport into leukocytes and fibroblasts.” Science 176, 1432.
Zurier, R. B., Hoffstein, S., and Weissmann, G. (1973). “Mechanisms of lysosomal enzyme release from human leukocytes. I. Effect of cyclic nucleotides and colchicine.” J. Cell Biol. 58, 27.
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Romeo, D., Zabucchi, G., Jug, M., Miani, N., Soranzo, M.R. (1975). Concanavalin A as A Probe for Studying the Mechanism of Metabolic Stimulation of Leukocytes. In: Chowdhury, T.K., Weiss, A.K. (eds) Concanavalin A. Advances in Experimental Medicine and Biology, vol 55. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-0949-9_15
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