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. 1974 Jun 1;139(6):1395–1414. doi: 10.1084/jem.139.6.1395

HORMONAL CONTROL OF LYSOSOMAL ENZYME RELEASE FROM HUMAN NEUTROPHILS

EFFECTS OF AUTONOMIC AGENTS ON ENZYME RELEASE, PHAGOCYTOSIS, AND CYCLIC NUCLEOTIDE LEVELS

L J Ignarro 1, T F Lint 1, W J George 1
PMCID: PMC2139682  PMID: 4364334

Abstract

The purpose of this investigation was to examine the effects of autonomic neurohormones, cyclic nucleotides, and related agents on the immunologic discharge of lysosomal enzymes from, and phagocytosis by, purified human neutrophils. In order to discern the possible intracellular mechanisms by which certain neurohormones influence neutrophil function, the concentrations of cyclic AMP and cyclic GMP in neutrophils were assessed during cell contact with phagocytizable particles and autonomic agents. The model system employed for study was the interaction of purified human neutrophils with rheumatoid arthritic (RA) serum-treated zymosan particles at 37°C in a neutral, balanced salt solution containing glucose. Neutrophils ingested the particles and discharged β-glucuronidase but not lactate dehydrogenase activity during 30 min of incubation. Treatment of zymosan particles with RA serum was more effective than treatment with normal serum with regard to the extent of both particle uptake and lysosomal enzyme release. During contact of neutrophils with RA serum-treated zymosan particles epinephrine, isoproterenol, and cyclic AMP inhibited both particle ingestion and β-glucuronidase discharge. These actions of epinephrine were associated with a concomitant elevation of cyclic AMP levels. In contrast to the actions of catecholamines and cyclic AMP, acetylcholine and cyclic GMP accelerated lysosomal enzyme release without affecting particle uptake. The actions of acetylcholine were associated with a concomitant elevation of cyclic GMP levels. Increases in neutrophil levels of cyclic GMP but not of cyclic AMP were associated also with the discharge of β-glucuronidase provoked by particles in the absence of added cholinergic agents. The data suggest that the immunologic release of lysosomal enzymes from human neutrophils can be regulated by autonomic neurohormones, perhaps via the selective formation of appropriate nucleotides.

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Selected References

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  1. Ali S. Y. The degradation of cartilage matrix by an intracellular protease. Biochem J. 1964 Dec;93(3):611–618. doi: 10.1042/bj0930611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bourne H. R., Lehrer R. I., Cline M. J., Melmon K. L. Cyclic 3',5'-adenosine monophosphate in the human lukocyte: synthesis, degradation, andeffects n neutrophil candidacidal activity. J Clin Invest. 1971 Apr;50(4):920–929. doi: 10.1172/JCI106564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown J. H., Mackey H. K., Riggilo D. A., Schwartz N. L. Studies on the acute inflammatory response. II. Influence of antihistaminics and catecholamines on formaldehyde-induced edema. J Pharmacol Exp Ther. 1968 Mar;160(1):243–248. [PubMed] [Google Scholar]
  4. Cochrane C. G., Aikin B. S. Polymorphonuclear leukocytes in immunologic reactions. The destruction of vascular basement membrane in vivo and in vitro. J Exp Med. 1966 Oct 1;124(4):733–752. doi: 10.1084/jem.124.4.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cox J. P., Karnovsky M. L. The depression of phagocytosis by exogenous cyclic nucleotides, prostaglandins, and theophylline. J Cell Biol. 1973 Nov;59(2 Pt 1):480–490. doi: 10.1083/jcb.59.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crowder J. G., Martin R. R., White A. Release of histamine and lysosomal enzymes by human leukocytes during phagocytosis of staphylococci. J Lab Clin Med. 1969 Sep;74(3):436–444. [PubMed] [Google Scholar]
  7. GIANETTO R., DE DUVE C. Tissue fractionation studies. 4. Comparative study of the binding of acid phosphatase, beta-glucuronidase and cathepsin by rat-liver particles. Biochem J. 1955 Mar;59(3):433–438. doi: 10.1042/bj0590433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. George W. J., Polson J. B., O'Toole A. G., Goldberg N. D. Elevation of guanosine 3',5'-cyclic phosphate in rat heart after perfusion with acetylcholine. Proc Natl Acad Sci U S A. 1970 Jun;66(2):398–403. doi: 10.1073/pnas.66.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. George W. J., Wilkerson R. D., Kadowitz P. J. Influence of acetylcholine on contractile force and cyclic nucleotide levels in the isolated perfused rat heart. J Pharmacol Exp Ther. 1973 Jan;184(1):228–235. [PubMed] [Google Scholar]
  10. Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldstein I. M., Brai M., Osler A. G., Weissmann G. Lysosomal enzyme release from human leukocytes: mediation by the alternate pathway of complement activation. J Immunol. 1973 Jul;111(1):33–37. [PubMed] [Google Scholar]
  12. Goldstein I., Hoffstein S., Gallin J., Weissmann G. Mechanisms of lysosomal enzyme release from human leukocytes: microtubule assembly and membrane fusion induced by a component of complement. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2916–2920. doi: 10.1073/pnas.70.10.2916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Götze O., Müller-Eberhard H. J. The c3-activator system: an alternate pathway of complement activation. J Exp Med. 1971 Sep 1;134(3):90–108. [PMC free article] [PubMed] [Google Scholar]
  14. Hawkins D., Cochrane C. G. Glomerular basement membrane damage in immunological glomerulonephritis. Immunology. 1968 May;14(5):665–681. [PMC free article] [PubMed] [Google Scholar]
  15. Hawkins D. Neutrophilic leukocytes in immunologic reactions: evidence for the selective release of lysosomal constituents. J Immunol. 1972 Feb;108(2):310–317. [PubMed] [Google Scholar]
  16. Henson E. C., Brunson J. G. Prevention of experimental allergic encephalomyelitis by combination of epinephrine and a phenothiazine derivative, propiomazine. Proc Soc Exp Biol Med. 1969 Jul;131(3):752–755. doi: 10.3181/00379727-131-33969. [DOI] [PubMed] [Google Scholar]
  17. Henson E. C., Brunson J. G. Studies of adjuvant-induced arthritis in the albino rat (CFN strain). Ann Rheum Dis. 1970 Mar;29(2):185–189. doi: 10.1136/ard.29.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Henson P. M. The immunologic release of constituents from neutrophil leukocytes. I. The role of antibody and complement on nonphagocytosable surfaces or phagocytosable particles. J Immunol. 1971 Dec;107(6):1535–1546. [PubMed] [Google Scholar]
  19. Hurd E. R., LoSpalluto J., Ziff M. Formation of leukocyte inclusions in normal polymorphonuclear cells incubated with synovial fluid. Arthritis Rheum. 1970 Nov-Dec;13(6):724–733. doi: 10.1002/art.1780130602. [DOI] [PubMed] [Google Scholar]
  20. Ignarro L. J., Colombo C. Enzyme release from polymorphonuclear leukocyte lysosomes: regulation by autonomic drugs and cyclic nucleotides. Science. 1973 Jun 15;180(4091):1181–1183. doi: 10.1126/science.180.4091.1181. [DOI] [PubMed] [Google Scholar]
  21. Ignarro L. J., Krassikoff N., Slywka J. Inhibition of lysosomal enzyme release by adenosine 5' triphosphate and N 6 ,O 2 -dibutyryl cyclic 3',5'-adenosine monophosphate. Life Sci I. 1972 Apr 1;11(7):317–322. doi: 10.1016/0024-3205(72)90057-4. [DOI] [PubMed] [Google Scholar]
  22. Ignarro L. J., Krassikoff N., Slywka J. Release of enzymes from a rat liver lysosome fraction: inhibition by catecholamines and cyclic3', 5'-adenosine monophosphate, stimulation by cholinergic agents and cyclic 3', 5'-guanosine monophosphate. J Pharmacol Exp Ther. 1973 Jul;186(1):86–99. [PubMed] [Google Scholar]
  23. Ignarro L. J. Neutral protease release from human leukocytes regulated by neurohormones and cyclic nucleotides. Nat New Biol. 1973 Oct 3;245(144):151–154. doi: 10.1038/newbio245151a0. [DOI] [PubMed] [Google Scholar]
  24. Ignarro L. J. Nonphagocytic release of neutral protease and beta-glucuronidase from human neutrophils. Regulation by autonomic neurohormones and cyclic nucleotides. Arthritis Rheum. 1974 Jan-Feb;17(1):25–36. doi: 10.1002/art.1780170106. [DOI] [PubMed] [Google Scholar]
  25. Ignarro L. J., Oronsky A. L., Perper R. J. Breakdown of noncollagenous chondromucoprotein matrix by leukocyte lysosome granule lysates from guinea pig, rabbit, and human. Clin Immunol Immunopathol. 1973 Nov;2(1):36–51. doi: 10.1016/0090-1229(73)90034-2. [DOI] [PubMed] [Google Scholar]
  26. Ignarro L. J., Slywka J. Changes in liver lysosome fragility, erythrocyte membrane stability, and local and systemic lysosomal enzyme levels in adjuvant-induced polyarthritis. Biochem Pharmacol. 1972 Mar 15;21(6):875–886. doi: 10.1016/0006-2952(72)90131-1. [DOI] [PubMed] [Google Scholar]
  27. Ignarro L. J., Slywka J., Krassikoff N. Inhibition of lysosomal enzyme release by catecholamines: possible mediation by cyclic 3',5'-adenosine monophosphate. Life Sci I. 1971 Nov 15;10(22):1309–1315. doi: 10.1016/0024-3205(71)90330-4. [DOI] [PubMed] [Google Scholar]
  28. Lichtenstein L. M., Margolis S. Histamine release in vitro: inhibition by catecholamines and methylxanthines. Science. 1968 Aug 30;161(3844):902–903. doi: 10.1126/science.161.3844.902. [DOI] [PubMed] [Google Scholar]
  29. Maillard J. L., Zarco R. M. Décomplémentation par un facteur extrait du venin de cobra. Effet sur plusieurs réactions immunes du cobaye et du rat. Ann Inst Pasteur (Paris) 1968 Jun;114(6):756–774. [PubMed] [Google Scholar]
  30. Manganiello V., Evans W. H., Stossel T. P., Mason R. J., Vaughan M. The effect of polystyrene beads on cyclic 3',5'-adenosine monophosphate concentration in leukocytes. J Clin Invest. 1971 Dec;50(12):2741–2744. doi: 10.1172/JCI106775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Müller-Eberhard H. J. Complement. Annu Rev Biochem. 1969;38:389–414. doi: 10.1146/annurev.bi.38.070169.002133. [DOI] [PubMed] [Google Scholar]
  32. Orange R. P., Austen W. G., Austen K. F. Immunological release of histamine and slow-reacting substance of anaphylaxis from human lung : I. Modulation by agents influencing cellular levels of cyclic 3',5'-adenosine monophosphate. J Exp Med. 1971 Sep 1;134(3):136–148. [PMC free article] [PubMed] [Google Scholar]
  33. Oronsky A., Ignarro L., Perper R. Release of cartilage mucopolysaccharide-degrading neutral protease from human leukocytes. J Exp Med. 1973 Aug 1;138(2):461–472. doi: 10.1084/jem.138.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. RABINOWITZ Y. SEPARATION OF LYMPHOCYTES, POLYMORPHONUCLEAR LEUKOCYTES AND MONOCYTES ON GLASS COLUMNS, INCLUDING TISSUE CULTURE OBSERVATIONS. Blood. 1964 Jun;23:811–828. [PubMed] [Google Scholar]
  35. SPECTOR W. G., WILLOUGHBY D. A. VASOACTIVE AMINES IN ACUTE INFLAMMATION. Ann N Y Acad Sci. 1964 Aug 27;116:839–846. doi: 10.1111/j.1749-6632.1964.tb52549.x. [DOI] [PubMed] [Google Scholar]
  36. Seyberth H. W., Schmidt-Gayk H., Jakobs K. H., Hackenthal E. Cyclic adenosine monophosphate in phagocytizing granulocytes and alveolar macrophages. J Cell Biol. 1973 May;57(2):567–571. doi: 10.1083/jcb.57.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Steiner A. L., Parker C. W., Kipnis D. M. Radioimmunoassay for cyclic nucleotides. I. Preparation of antibodies and iodinated cyclic nucleotides. J Biol Chem. 1972 Feb 25;247(4):1106–1113. [PubMed] [Google Scholar]
  38. Stossel T. P., Mason R. J., Hartwig J., Vaughan M. Quantitative studies of phagocytosis by polymorphonuclear leukocytes: use of emulsions to measure the initial rate of phagocytosis. J Clin Invest. 1972 Mar;51(3):615–624. doi: 10.1172/JCI106851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tew J. G., Hess W. M., Donaldson D. M. Lysozyme and beta-lysin release stimulated by antigen-antibody complexes and bacteria. J Immunol. 1969 Mar;102(3):743–750. [PubMed] [Google Scholar]
  40. Tolone G., Bonasera L., Bruno R. The effects of adrenergic blocking agents on passive cutaneous anaphylaxis reactions in the rat. Pathol Microbiol (Basel) 1971;37(3):194–200. doi: 10.1159/000162321. [DOI] [PubMed] [Google Scholar]
  41. Vallota E. H., Müller-Eberhard H. J. Formation of C3a and C5a anaphylatoxins in whole human serum after inhibition of the anaphylatoxin inactivator. J Exp Med. 1973 May 1;137(5):1109–1123. doi: 10.1084/jem.137.5.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weissmann G., Dukor P., Zurier R. B. Effect of cyclic AMP on release of lysosomal enzymes from phagocytes. Nat New Biol. 1971 Jun 2;231(22):131–135. doi: 10.1038/newbio231131a0. [DOI] [PubMed] [Google Scholar]
  43. Weissmann G. The role of lysosomes in inflammation and disease. Annu Rev Med. 1967;18:97–112. doi: 10.1146/annurev.me.18.020167.000525. [DOI] [PubMed] [Google Scholar]
  44. Weissmann G., Zurier R. B., Hoffstein S. Leukocytic proteases and the immunologic release of lysosomal enzymes. Am J Pathol. 1972 Sep;68(3):539–564. [PMC free article] [PubMed] [Google Scholar]
  45. Weissmann G., Zurier R. B., Spieler P. J., Goldstein I. M. Mechanisms of lysosomal enzyme release from leukocytes exposed to immune complexes and other particles. J Exp Med. 1971 Sep 1;134(3):149–165. [PMC free article] [PubMed] [Google Scholar]
  46. Wright D. G., Malawista S. E. The mobilization and extracellular release of granular enzymes from human leukocytes during phagocytosis. J Cell Biol. 1972 Jun;53(3):788–797. doi: 10.1083/jcb.53.3.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. ZIFF M., GRIBETZ H. J., LOSPALLUTO J. Effect of leukocyte and synovial membrane extracts on cartilage mucoprotein. J Clin Invest. 1960 Feb;39:405–412. doi: 10.1172/JCI104051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zurier R. B., Hoffstein S., Weissmann G. Cytochalasin B: effect on lysosomal enzyme release from human leukocytes. Proc Natl Acad Sci U S A. 1973 Mar;70(3):844–848. doi: 10.1073/pnas.70.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]

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