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Modulation of phagocytosis and intracellular bactericidal activity of polymorphonuclear and mononuclear cells by cationic proteins from human granulocytes: Alternative pathway of phagocytic enhancement

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Abstract

Cationic lysosomal proteins from human polymorphonuclears (PMN) were isolated by column chromatography and divided into five fractions. On acrylamide gel electrophoresis, fraction I had four bands slower than lysozyme (LZM) mobility; fraction II had five or six bands slower than LZM; fraction III had at least seven bands slower and two bands faster than LZM; fraction IV contained LZM, two bands faster and a few faint bands slower than LZM; fraction V was composed of almost pure LZM. Partial characterization of the fractions showed presence of neutral protease in fractions I-IV, chymotrypsin in fraction III, lysozyme in fractions IV and V, and phospholipase A2 mainly in fractions II and III. Modulatory activity of fractions I-V were tested at concentrations up to 50 μg/ml. Enhancement of phagocytosis ofStaphylococcus aureus was observed by fractions I, IV, and V, whereas phagocytic index was enhanced by all but the fraction II. Intracellular bactericidal activity (ICBA) was markedly enhanced by fractions I, II, and V. Addition of DNA or cytochalasine B inhibited or abolished phagocytosis-enhancing activity of cationic fractions. Their influence on ICBA was much less pronounced. Fraction III enhanced phagocytic index and phagocytosis ofE. colt, whereas fractions I and II enhanced intracellular bactericidal activity against this bacteria. Enhancement of phagocytic activity of monocytes has also been observed. The data suggest that some cationic lysosomal fractions from human PMNs enhance phagocytosis and phagocytic index by human PMNs and monocytes and intracellular bactericidal activity of human PMNs. This alternative pathway of phagocytic enhancement is unrelated to the previously described enhancers of phagocytosis and may play a role in defense mechanisms against infection.

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References

  1. Wardlaw, A. C. 1962. The complement-dependent bacteriolytic activity of normal human serum. I. The effect of pH and ionic strength and the role of lysozyme.J. Exp. Med. 115:1231–1249.

    Google Scholar 

  2. Zeya, H. I., andJ. K. Spitznagel. 1966. Antimicrobial specificity of leukocyte lysosomal cationic proteins.Science 154:1049–1051

    Google Scholar 

  3. Welsh, I. R. H., andJ. K. Spitznagel, 1971. Distribution of iysosomal enzymes, cationic proteins and bactericidal substances in subcellular fractions of human polymorphonuclear leukocytes.Infect. Immun. 4:97–102.

    Google Scholar 

  4. Zeya, H. I., andJ. K. Spitznagel. 1966. Cationic proteins of polymorphuclear leukocyte lysosomes, I. Resolution of antibacterial and enzymatic activities.J. Bacteriol. 91:750–754.

    Google Scholar 

  5. Stossel, T. P. 1975. Phagocytosis: Recognition and ingestion.Semin. Hematol. 12:83–116.

    Google Scholar 

  6. Gordon, S., J. Todd, andZ. A. Cohn. 1974. In vitro synthesis and secretion of lysozyme by mononuclear phagocytes.J. Exp. Med. 139:1228–1248.

    Google Scholar 

  7. Ranadive, N. S., andC. G. Cochrane. 1968. Isolationand characterization of permeability factors from rabbit neutrophils.J. Exp. Med. 128:605–622.

    Google Scholar 

  8. Janoff, A., andB. W. Zweifach. 1964. Adhesion and emigration of leukocytes produced by cationic proteins of lysosomes.Science 144:1456–1458.

    Google Scholar 

  9. Janoff, A, andB. W. Zweifach. 1964. Production of inflammatory changes in the microcirculation by cationic proteins extracted from lysosomes.J. Exp. Med. 120:747–764.

    Google Scholar 

  10. Golub, E. S., andJ. K. Spitznagel. 1965. The role of lysosomes in hypersensitivity reactions: Tissue damage by polymorphonuclear neutrophil lysosomes.J. Immunol. 95:1060–1066.

    Google Scholar 

  11. Janoff, A. S. Schaefer, andJ. Scherer. 1965. Mediators of inflammation in leukocyte lysosomes. II. Mechanism of action of lysosomal cationic proteins upon vascular permeability in the rat.J. Exp. Med. 122:841–851.

    Google Scholar 

  12. Olsson, I., andP. Venge, 1974. Cationic proteins of human granulocytes. II. Separation of the cationic proteins of the granules of leukaemic myeloid cells.Blood 44:235–246.

    Google Scholar 

  13. Olsson, I., andP. Venge. 1972. Cationic proteins of human granulocytes. I. Isolation of the cationic proteins from the granules of leukaemic myeloid cells.Scand. J. Haematl. 9:204–214.

    Google Scholar 

  14. Zeya, H. I., andJ. K. Spitznagel. 1968. Arginine-rich proteins of polymorphonuclear leukocyte lysosomes. Antimicrobial specificity and biochemical heterogeneity.J. Exp. Med. 127:927–941.

    Google Scholar 

  15. Spitznagel, J. K., F. G. Dalldorf, M. S. Leffell, J. D. Folds, I. R. H. Welsh, M. H. Cooney, andL. E. Martin. 1974. Character of azurophil and specific granules purified from human polymorphonuclear leukocytes.Lab. Invest. 30:774–785.

    Google Scholar 

  16. Zeya, H. I., andJ. K. Spitznagel. 1966. Cationic proteins of polymorphonuclear leukocyte lysosomes. II. Composition, properties and mechanism of antibacterial action.J. Bacteriol. 91:755–762.

    Google Scholar 

  17. Zeya, H. I., andJ. K. Spitznagel. 1963. Antibacterial and enzymic basic proteins from leukocyte lysosomes: Separation and identification.Science 142:1085–1087.

    Google Scholar 

  18. Pruzanski, W., andS. Saito. 1978. The influence of natural and synthetic cationic substances on phagocytic activity of human polymorphonuclear cells. An alternative pathway of phagocytic enhancement.Exp. Cell Res. 117:1–13.

    Google Scholar 

  19. Ranadive, N. S., A. N. Sajnani, K. Alimurka, andH. Z. Movat. 1973. Release of basic proteins and lysosomal enzymes from neutrophil leukocytes of the rabbit.Int. Arch. Allergy 45:880–898.

    Google Scholar 

  20. Gordon, J., andF. C. Thompson. 1937. Artificial opsonization of bacteria.Br. J. Exp. Pathol 18:390–395.

    Google Scholar 

  21. Westwood, F. R., andE. Longstaff. 1976. Stimulation of cellular ingestion by basic proteins in vitro.Br. J. Cancer 33:392–399.

    Google Scholar 

  22. Falke, D., E. M. Lemmel, L. S. Richardson, andH. U. Wolf. 1975. Enhancement of phagocytosis by compound 48/80 and its influence upon Ca+ dependent ATP-ase.Arzneimittelforschung 25:1258–1261.

    Google Scholar 

  23. Devries, A., J. Salgo, Y. Matoth, A. Nevo, andE. Katchalski. 1955. Effect of basic polyamino acids on phagocytosis in vitro.Arch. Int. Pharmacody. 104:1–10.

    Google Scholar 

  24. Hallgren, R., andP. Venge. 1976. Cationic proteins of human granulocytes: Enhancement of phagocytosis of staphylococcus protein A-IgG complexes.Inflammation 1:237–246.

    Google Scholar 

  25. Gordon, L. I., S. D. Douglas, N. E. Kav, E. F. Osserman, H. S. Jacob. 1979. Modulation of neutrophil function by lysozyme. Potential negative feedback system of inflammation.J. Clin. Invest. 64:226–232.

    Google Scholar 

  26. Boyum, A. 1968. Isolation of mononuclear cells and granulocytes from human blood.Scand. J. Clin. Lab. Invest. Suppl. 97.21:77.

    Google Scholar 

  27. Kaplow, L. S., andC. Ladd. 1965. Simplified myeloperoxidase stain using benzidine dihydrochloride.Blood 26:215–219.

    Google Scholar 

  28. Yam, L. T., C. Y. Li, andW. H. Crosby. 1971. Cytochemical identification of monocytes and granulocytes.Am. J. Clin. Pathol. 55:283–290.

    Google Scholar 

  29. Williams, C. A. 1968. Determination of nitrogen by Nessler reaction.In Methods in Immunology and Lmmunochemstry. C. A. Williams and M. W. Chase (ed). Vol. 2, Academic Press, New York. 266–270.

    Google Scholar 

  30. Fishman, W. M. 1963. Determination of β-glucuronidase.In Methods of Enzymatic Analysis. H. U. Bergmeyer, editor. Academic Press, New York. 869.

    Google Scholar 

  31. Starkey, P. M., andA. J. Barrett. 1976. Neutral proteinases of human spleen. Purification and criteria for homogeneity of elastases and cathepsin G.Biochem. J. 155:255–263.

    Google Scholar 

  32. Osserman, E. F., andD. P. Lawlor. 1966. Serum and urinary lysozyme (muramidase) in monocytic and myelomonocytic leukemia.J. Exp. Med. 124:921–951.

    Google Scholar 

  33. Pruzanski, W., andM. E. Platts. 1970. Serum and urinary proteins, lysozyme (muramidase), and renal dysfunction in mono- and myelomonocytic leukemia.J. Clin. Invest. 49:1694–1708.

    Google Scholar 

  34. Worthington Manual. 1967. Peroxidase 1.11.1.17. Worthington Chemical Corp. Freehold, New Jersey,

  35. Christman, J. K., S. C. Silverstein, andG. Acs. 1977. Fibrin plate method.In Proteinases in Mammalian Cells and Tissues. A. J. Barrett, editior. North Holland, Amsterdam. 136.

    Google Scholar 

  36. Ranadive, N. S., andC. G. Cochrane, 1971. Mechanism of histamine release from rat mast cells by cationic proteins from neutrophil lysosomes.J. Immunol. 106:506–516.

    Google Scholar 

  37. Vadas, P., andJ. B. Hay. 1980. The release of phospholipase A2 from aggregated platelets and stimulated macrophages of sheep.Life Sci. 26:1721–1729.

    Google Scholar 

  38. Peterson, P. K., J. Verhoef, L. D. Sabath, andP. G. Quie. 1976. Extracellular and bacterial factors influencing staphylococcal phagocytosis and killing by human polymorphonuclear leukocytes.Infect. Immun. 14:496–501.

    Google Scholar 

  39. Zimmerman, H. J., andJ. B. Henry. 1969.In Clinical Diagnosis by Laboratory Methods. 14th ed. I. Davidsohn and J. B. Henry, editors. W. B. Saunders, Philadelphia. 725.

    Google Scholar 

  40. Tan, J. S., C. Watanakunakorn, andJ. P. Phair. 1971. A modified assay of netrophil function: Use of lysostaphin to differentiate defective phagocytosis from impaired intracellular killing.J. Lab. Clin. Med. 78:316–322.

    Google Scholar 

  41. Goldner, M., H. Farkas-Himsley, A. Kormendy, andM. Skinner. 1983. Bacterial phagocytosis monitored by fluorescence and extracellular quenching: Ingestion and intracellular killing.Lab. Med. 14:291–294.

    Google Scholar 

  42. Pruzanski, W., andK. Delmage. 1977, Cytotoxic and eytolytic activity of homogeneous cold agglutinins on peripheral blood monocytes.Clin. Immunol. Immunopaihol. 7:130–138.

    Google Scholar 

  43. Stossel, T. P. 1974. Phagocytosis.N. Engl. J. Med. 290:717–723.

    Google Scholar 

  44. Windhorst, D. B. 1970. Functional defects of neutrophiles.Adv. Inter. Med. 16:329–349.

    Google Scholar 

  45. Van Oss, C. J., C. F. Gillman, andR. J. Good. 1972. The influence of the shape of phagocytes on their adhesiveness.Immunol. Commun. 1:627–636.

    Google Scholar 

  46. Karnovsky, M. L. 1962. Metabolic basis of phagocytic activity.Physiol. Rev. 42:143–168.

    Google Scholar 

  47. Sbarra, A. J., A. A. Jacobs, R. R. Strauss, B. B. Paul, andG. W. Mitchell, Jr. 1971. The biochemical and antimicrobial activities of phagocytizing cells.Am. J. Clin. Nutr. 24:272–281.

    Google Scholar 

  48. Cohn, Z. A., andJ. G. Hirsch. 1960. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes.J. Exp. Med. 112:983–1004.

    Google Scholar 

  49. Weissmann, G., andP. Dukor. 1970. The role of lysosomes in immune responses.Adv. Immunol. 12:283–331.

    Google Scholar 

  50. Weissmann, G. R. B. Zurier, P. J. Spieler, and1. M. Goldstein. 1971. Mechanism of lysosomal enzyme release from leukocytes exposed to immune complexes and other particles,J. Exp. Med. 134:149s-165s.

    Google Scholar 

  51. Clark, R. A., I. Olsson, andS. J. Klebanoff. 1976, Cytotoxicity for tumor cells of cationic proteins from human neutrophil granules.J. Cell. Biol. 70:719–723.

    Google Scholar 

  52. Baker, P. J., T. F. Lint, J. Siegel, M. W. Kies, andH. Gewurz. 1976. Potentiation of C5,6-initiated lysis by leukocyte cationic proteins, myelin basic proteins and lysine-rich histones.Immunology 30:467–473.

    Google Scholar 

  53. Saba, H. I., H. R. Roberts, andJ. C. Herion. 1967. The anticoagulant activity of lysosomal cationic proteins from polymorphonuclear leukocytes.J. Clin. Invest. 46:580–589.

    Google Scholar 

  54. Klebanoff, S. J. 1970. Myeloperoxidase: Contribution to the microbicidal activity of intact leukocytes.Science 169:1095–1097.

    Google Scholar 

  55. Modrzakowski, M. C., M. H. Cooney, L. E. Martin, andJ. K. Spitznagel. 1979. Bactericidal activity of fractionated granule contents from human polymorphonuclear leukocytes.Infect. Immun. 23:587–591.

    Google Scholar 

  56. Odeberg, H., andI. Olsson. 1975. Antibacterial activity of cationic proteins from human granulocytes.J. Clin. Invest. 56:1118–1124.

    Google Scholar 

  57. Venge, P., andI. Olsson. 1975. Cationic proteins of human granulocytes. VI. Effects on the complement system and mediation of chemotactic activity.J. Immunol. 115:1505–1508.

    Google Scholar 

  58. Steigbigel. R. T., L. H. Lambert, Jr., andJ. S. Remington. 1974. Phagocytic and bactericidal properties of normal human monocytes.J. Clin. Invest. 53:131–142.

    Google Scholar 

  59. Lemarbre, P., J. Rinehart, N. Kay, E. Osserman, andH. S. Jacob. 1979. Lysozyme (LYZ) enhances monocyte-mediated tumoricidal activity; A potential beneficial amplifier in tumor immunity.Blood (Suppl 1)54:103a.

    Google Scholar 

  60. Tischendorf, F. W. G. Ledderose, W. Wilmanns, M. M. Tischendorf, andG. W. Tischendorf. 1973. Cationic leukocyte antigen in urine of patients with chronic myelocytic leukaemia.Nature 245:379–380.

    Google Scholar 

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Authors and Affiliations

  1. Immunology Diagnostic & Research Centre Departments of Medicine and Pathology, The Wellesley Hospital, University of Toronto, Toronto, Ontario, Canada

    W. Pruzanski, N. S. Ranadive & S. Saito

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  1. W. Pruzanski
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  2. N. S. Ranadive
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  3. S. Saito
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Supported by grants-in-aid from the Medical Reseach Council of Canada and the Wellesley Hospital Research Institute.

This study was presented in part at the 9th International RES Congress, Davos, Switzerland, 1982.

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Pruzanski, W., Ranadive, N.S. & Saito, S. Modulation of phagocytosis and intracellular bactericidal activity of polymorphonuclear and mononuclear cells by cationic proteins from human granulocytes: Alternative pathway of phagocytic enhancement. Inflammation 8, 445–457 (1984). https://doi.org/10.1007/BF00918219

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