Abstract
Human blood leukocytes and platelets and mouse peritoneal macrophages emit very rapid and very intense Luminol-dependent chemiluminescence (CL) signals when treated with streptococci, staphylococci, or with zymosan, which have been preopsonized with arginine-rich histone, dextran sulfate or polyanetholesulfonate (liquoid). Liquoid alone at 10–30μg/2×105 leukocytes also triggers intense CL responses in the absence of a carrier. Strong CL can also be triggered, and at the same levels, when the various polyelectrolytes are simply mixed with the bacteria or zymosan and added to the leukocyte suspensions. The CL responses induced by the polyelectrolyte-bacteria complexes greatly exceed those triggered in leukocytes by antibody-complement-coated particles. Liquoid also shows a unique property of markedly augmenting CL signals which have already been induced by other ligand-coated bacteria or zymosan particles. Streptococci and staphylococci were found to be much superior to zymosan, Gram-positive bacilli, orE. coli as carriers for the various polyelectrolytes in the CL reaction. Neither protamine sulfate, lysozyme, myeloperoxidase, crystalline ribonuclease (all cationic in nature), chondroitin sulfate, heparin, nor alginate sulfate acted as ligands for triggering CL, when used to opsonize bacteria or zymosan. The induction of CL in blood leukocytes by the various ligand-coated bacteria is markedly inhibited by azide, KCN catalase, aminotriazole, and EDTA, agents known to inhibit the production of oxygen radicals following stimulation of leukocytes by opsonized bacteria. Two children diagnosed for chronic granulomatous diseases (CGD) of childhood and an apparently healthy sister of one of the male patients completely failed to respond with CL either to the polyelectrolyte-bacteria complexes, liquoid or antibody-coated bacteria and zymosan. It is proposed that liquoid be employed for the rapid screening of defects in certain oxygen-dependent metabolic processes in both PMNs and macrophages. It is also suggested that polyelectrolytes like the ones described in this study may markedly enhance the bactericidal properties of leukocytes and macrophages towards both extracellular and intracellular microorganisms and may perhaps also augment the tumoricidal effects of activated macrophages.
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Trush, M. A., M. E. Wilson, andK. Van Dyke. 1978. The generation of chemiluminescence (CL) by phagocytic cells.Methods Enzymol. 57:462–494.
Allen, R. C., R. L. Stjernholm, andR. H. Steele. 1972. Evidence for the generation of an electronic excitation state(s) in human polymorphonuclear leukocytes and its participation in bactericidal activity.Biochem. Biophys. Res. Commun. 47:679–684.
Babior, B. M., R. C. Kipnes, andJ. T. Curnutte. 1973. Biological defence mechanisms: The production by leukocytes of superoxide, a potential bactericidal agent.J. Clin. Invest. 53:741–744.
Klebanoff, S. J.. 1975. Antimicrobial mechanisms in neutrophilic polymorphonuclear leukocytes.Semin. Hematol. 12:117–142.
Allen, R. C., andL. D. Loose. 1976. Phagocytic activation of a Luminol-dependent chemiluminescence in rabbit alveolar and peritoneal macrophages.Biochem. Biophys. Res. Commun. 69:245–252.
Root, R. K., andM. S. Cohen. 1981. The microbicidal mechanisms of human neutrophils and eosinophils.Rev. Infect. Dis. 3:565–598.
Stjernholm, R. L., R. C. Allen, R. H. Steele, W. W. Waring, andJ. A. Harris. 1973. Impaired chemiluminescence during phagocytosis of opsonized bacteria.Infect. Immun. 7:313–314.
Stevens, P., andL. S. Young. 1977. Quantitative granulocyte chemiluminescence in the rapid detection of impaired opsonization ofEscherichia coli. Infect. Immun.16:796–804.
Allen, R. C. 1977. Evaluation of serum opsonic capacity by quantitating the initial chemiluminescence response from phagocytozing polymorphonuclear leukocytes.Infect. Immunol. 15:828–833.
Yanai, M., andP. G. Quie. 1981. Chemiluminescence by polymorphonuclear leukocytes adhering to surfaces.Infect. Immun. 32:1181–1186.
Muller, S., S. Falkenberg, R. A. Fromtling, A. M. Feomtling, andV. Klimetzek. 1981. Signals of chemiluminescence emitted by spleen cells and bone marrow macrophages after stimulation with mitogens and particular substance.In Bioluminescence and Chemiluminescence. M. A. DeLuca, W. O. McElroy, editors. Academic Press, New York. 721–727.
Ginsburg, I., M. N. Sela, A. Morag, Z. Ravid, Z. Duchan, M. Ferne, S. Rabinowitz, P. Page-Thomas, P. Davies, J. Niccols, J. Humes, andR. Bonney. 1981. Role of leukocyte factors and cationic polyelectrplytes in phagocytosis of group A streptococci andCandida albicans by neutrophils, macrophages, fibroblasts and epithelial cells: Modulation by anionic polyelectrolytes in relation to the pathogenesis of chronic inflammation.Inflammation 5:289–312.
Ginsburg, I., M. Lahav, M. Ferne, andS. Muller. 1982. Cationic polyelectrolytes and leukocyte factors function as opsonins, as triggers of chemiluminescence and as activators of autolytic systems in bacteria: Modulation by anionic polyelectrolytes in relation to inflammation.In Macrophages and Natural Killer Cells: Regulation and Function. Plenum, New York. 151–160.
Ginsburg, I., andP. G. Quie. 1980. Modulation of human polymorphonuclear leukocyte chemotaxis by leukocyte extracts, bacterial products, inflammatory exudates and polyelectrolytes.Inflammation 4:301–311.
Lahav, M., andI. Ginsburg. 1977. Effect of leukocyte hydrolases on bacteria. X. The role played by leukocyte factors, cationic polyelectrolytes and by membrane-damaging agents in the lysis ofStaphylococcus aureus. Relation to chronic inflammatory processes.Inflammation 2:165–177.
Ginsburg, I., J. Goultchin, A. Stabholtz, N. Ne'eman, M. Lahav, L. Landstrom, andP. G. Quie. 1980. Streptococcal and staphylococcal arthritis: Can chronic arthritis in the human be caused by highly chemotactic degradation products generated from bacteria by leukocyte enzymes and by the deactivation of leukocytes by inflammatory exudates, polyelectrolytes, leukocyte hydrolases and by cell sensitizing agents derived from bacteria?In Trends in Inflammation Research, Vol. 1. G. P. Velo, editor. Birkhauser Verlag, Basel. 260–270.
Easmon, C. S. F., P. J. Cole, A. J. Williams, andM. Hastings. 1980. The measurement of opsonic and phagocytic function by luminol-dependent chemiluminescence.Immunology 41:67–74.
Roos, D. 1980. The metabolic response to phagocytosis.In The Cell Biology of Inflammation. G. Weissmann, editor. Elsevier/North Holland Biomedical Press, Amsterdam. 377–385.
Rossi, F., D. Romeo, andP. Patriarca. 1972. Mechanism of phagocytosis-associated oxidative metabolism in polymorphonuclear leukocytes and macrophages.J. Reticuloendothel. Soc. 12:127–149.
Johnston, R., Jr., C. A. Godzik, andZ. A. Cohn. 1978. Increased superoxide anion production by immunologically activated and chemically elicited macrophages.J. Exp. Med. 148:115–127.
Dechatelet, L. R. 1978. Initiation of the respiratory burst in human polymorphonuclear neutrophils: A critical review.J. Reticuloendothel. Soc. 24:73–91.
Allgower, M. 1947. Uber di wirkung non heparin, polyanetholsulfosauren natrium (Liquoid Roche) and tribasischen natriumcitrat auf menchlische leukozyten in vitro.Schwiez. Med. Mochenschr. 77:40–43.
Belding, M. E., andS. J. Klebanoff. 1972. Effect of sodium polyanetholesulfonate on antimicrobial system in blood.Appl. Microbiol. 24:691–698.
Klebanoff, S. J., andR. A. Clark. 1977. Iodination by human polymorphonuclear leukocytes: A re-evaluation.J. Lab. Clin. Med. 89:675–686.
DeVries, A., J. Salgo, Y. Mathot, A. Nevo, andE. Katchalski. 1955. The effect of polyamino acids on phagocytosis in vitro.Arch. Int. Pharmacodyn. 104:1–10.
Ginsburg, I., P. Chkistensen, I. Eliasson, andC. Schalen. 1982. Cationic polyelectrolytes, liquoid and leukocyte extracts modulate the binding of IgG to group A streptococcal Fc-receptors.Acta Pathol. Microbiol. Scand., Sec. B. 90:161–168.
Ginsburg, I., andM. Lahav. 1982. The effect of leukocyte hydrolases on bacteria. XVI. The activation by leukocyte factors and cationic substances of autolytic enzymes inStaphylococcus aureus: Modulation by anionic polyelectrolytes in relation to survival of bacteria in inflammatory exudates.Inflammation 6:269–284.
Zeya, H. I., andJ. K. Spitznagel. 1968. Arginine-rich proteins of polymorphonuclear leukocyte lysosome: Antimicrobial specificity and biochemical heterogeneity.J. Exp. Med. 127:927–941.
Ginsburg, I. 1982. The role of leukocyte and serum factors and of cationic polyelectrolytes in the lysis and biodegradation ofStaphylococcus aureus: Relation to the pathogenesis of staphylococci infections.In Staphylococci and Staphylococcal Infections. J. H. Adlam and C. S. F. Easmon, editors. Academic Press, London (in press).
Klebanoff, S. J., andR. A. Clark. 1978. The Neutrophil: Function and Clinical Disorders. North Holland, Amsterdam.
Weissmann, G., J. E. Smolen, andH. M. Korchak. 1980. Release of inflammatory mediators from stimulated neutrophils.N. Engl. J. Med. 303:27–34.
Elsbach, P. 1980. Degradation of microorganisms by phagocytic cells.Rev. Infect. Dis. 2:106–128.
Zeya, H. I., andJ. P. Spitznagel. 1966. Cationic proteins of polymorphonuclear leukocyte lysosomes. II. Composition, properties and mechanisms of antibacterial action,J. Bacteriol. 91:755–762.
Ginsburg, I. 1979. The role of lysosomal factors of leukocytes in the biodegradation and storage of microbial constituents in infectious granulomas.In Lysosomes in Applied Biology and Therapeutics, Vol. 6, J. T. Dingle, P. J. Jacques, and I. H. Shaw, North Holland Publishing Company, Amsterdam. 327–406.
Neeman, N., N. M. Sela, S. Chanes, L. Bierkenfeld, D. Kutani, M. Lahav, andI. Ginsburg. 1979. The effect of leukocyte hydrolases on bacateria XIV. Bacteriolytic effects of human sera, synovial fluids and purulent exudates onStaphylococcus aureus andStreptococcus faecalis: Modulation by Cohn's fraction II and by polyelectrolytes.Inflammation 3:379–394.
Neeman, N., Z. Duchan, M. Lahav, M. N. Sela, andI. Ginsburg. 1976. The effect of leukocyte hydrolases on bacteria. VII. Bactericidal and bacteriolytic reactions mediated by leukocytes and tissue extracts and their modulation by polyelectrolytes.Inflammation 1:261–284.
Olsson, I., andP. Venge. 1972. Cationic proteins of human granulocytes. I. Isolation of the cationic proteins from the granules of leukemic myeloid cells.Scand. J. Haematol. 9: 204–214.
Murray, H. W., andZ. A. Cohn. 1980. Macrophage oxygen-dependent antimicrobial activity. III. Enhanced oxidative metabolism as an expression of macrophage activation.J. Exp. Med. 152:1596–1609.
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This study was supported by a research grant obtained from Dr. Samuel Robbins of Cleveland, Ohio, who also kindly contributed the LKB-Chemiluminescence Instrument; by a grant from the Chief Scientist, Ministry of Health, Government of Israel; and by a grant in honor of Dr. Maurice D. Turbow of Los Angeles by his friends—members of the Alpha Omega Fraternity, Los Angeles Alumni Chapter.
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Ginsburg, I., Borinsky, R., Lahav, M. et al. Bacteria and zymosan opsonized with histone, dextran sulfate, and polyanetholesulfonate trigger intense chemiluminescence in human blood leukocytes and platelets and in mouse macrophages. Inflammation 6, 343–364 (1982). https://doi.org/10.1007/BF00917306
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DOI: https://doi.org/10.1007/BF00917306