Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1973 Aug 1;58(2):249–264. doi: 10.1083/jcb.58.2.249

SEQUENTIAL DEGRANULATION OF THE TWO TYPES OF POLYMORPHONUCLEAR LEUKOCYTE GRANULES DURING PHAGOCYTOSIS OF MICROORGANISMS

Dorothy Ford Bainton 1
PMCID: PMC2109046  PMID: 4729503

Abstract

The sequential discharge of neutrophilic polymorphonuclear leukocyte (PMN) granules—azurophils and specifics—was investigated by electron microscopy and cytochemistry. Thus the enzyme content of PMN phagocytic vacuoles was determined at brief intervals after phagocytosis of bacteria, utilizing peroxidase as a marker enzyme for azurophil granules, and alkaline phosphatase for specifics. At 30 s, approximately half the phagocytic vacuoles were reactive for alkaline phosphatase, whereas none contained peroxidase. Peroxidase-containing vacuoles were rarely seen at 1 min, but by 3 min, vacuoles containing both enzymes were consistently present. Alkaline phosphatase was found in both small and large vacuoles, whereas peroxidase was visible only in large ones. By 10 min, very big phagocytic vacuoles containing considerable amounts of reaction product for both enzymes were evident. These observations indicate that the two types of PMN granules discharge in a sequential manner, specific granules fusing with the vacuole before azurophils. In an earlier paper, we reported that the pH of phagocytic vacuoles drops to 6.5 within 3 min and to ∼4 within 7–15 min. Substances known to be present in specific granules (alkaline phosphatase, lysozyme, and lactoferrin) function best at neutral or alkaline pH, whereas most of those contained in azurophil granules (i.e., peroxidase and the lysosomal enzymes) have pH optima in the acid range. Hence the sequence of granule discharge roughly parallels the change in pH, thereby providing optimal conditions for coordinated activity of granule contents.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baggiolini M., De Duve C., Masson P. L., Heremans J. F. Association of lactoferrin with specific granules in rabbit heterophil leukocytes. J Exp Med. 1970 Mar 1;131(3):559–570. doi: 10.1084/jem.131.3.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baggiolini M., Hirsch J. G., De Duve C. Further biochemical and morphological studies of granule fractions from rabbit heterophil leukocytes. J Cell Biol. 1970 Jun;45(3):586–597. doi: 10.1083/jcb.45.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baggiolini M., Hirsch J. G., De Duve C. Resolution of granules from rabbit heterophil leukocytes into distinct populations by zonal sedimentation. J Cell Biol. 1969 Feb;40(2):529–541. doi: 10.1083/jcb.40.2.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. I. Histochemical staining of bone marrow smears. J Cell Biol. 1968 Nov;39(2):286–298. doi: 10.1083/jcb.39.2.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. II. Cytochemistry and electron microscopy of bone marrow cells. J Cell Biol. 1968 Nov;39(2):299–317. doi: 10.1083/jcb.39.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bainton D. F., Farquhar M. G. Origin of granules in polymorphonuclear leukocytes. Two types derived from opposite faces of the Golgi complex in developing granulocytes. J Cell Biol. 1966 Feb;28(2):277–301. doi: 10.1083/jcb.28.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. COHN Z. A., HIRSCH J. G. The influence of phagocytosis on the intracellular distribution of granule-associated components of polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:1015–1022. doi: 10.1084/jem.112.6.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. COHN Z. A., HIRSCH J. G. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:983–1004. doi: 10.1084/jem.112.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. COHN Z. A., MORSE S. I. Functional and metabolic properties of polymorphonuclear leucocytes. I. Observations on the requirements and consequences of particle ingestion. J Exp Med. 1960 May 1;111:667–687. doi: 10.1084/jem.111.5.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. COHN Z. A. The fate of bacteria within phagocytic cells. I. The degradation of isotopically labeled bacteria by polymorphonuclear leucocytes and macrophages. J Exp Med. 1963 Jan 1;117:27–42. doi: 10.1084/jem.117.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Duve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol. 1966;28:435–492. doi: 10.1146/annurev.ph.28.030166.002251. [DOI] [PubMed] [Google Scholar]
  12. Elsbach P., Patriarca P., Pettis P., Stossel T. P., Mason R. J., Vaughan M. The appearance of lecithin- 32 P, synthesized from lysolecithin- 32 P, in phagosomes of polymorphonuclear leukocytes. J Clin Invest. 1972 Jul;51(7):1910–1914. doi: 10.1172/JCI106994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farquhar M. G., Bainton D. F., Baggiolini M., de Duve C. Cytochemical localization of acid phosphatase activity in granule fractions from rabbit polymorphonuclear leukocytes. J Cell Biol. 1972 Jul;54(1):141–156. doi: 10.1083/jcb.54.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  15. HIRSCH J. G., COHN Z. A. Degranulation of polymorphonuclear leucocytes following phagocytosis of microorganisms. J Exp Med. 1960 Dec 1;112:1005–1014. doi: 10.1084/jem.112.6.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HIRSCH J. G. Cinemicrophotographic observations on granule lysis in polymorphonuclear leucocytes during phagocytosis. J Exp Med. 1962 Dec 1;116:827–834. doi: 10.1084/jem.116.6.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. HORN R. G., SPICER S. S., WETZEL B. K. PHAGOCYTOSIS OF BACTERIA BY HETEROPHIL LEUKOCYTES: ACID AND ALKALINE PHOSPHATASE CYTOCHEMISTRY. Am J Pathol. 1964 Aug;45:327–335. [PMC free article] [PubMed] [Google Scholar]
  18. Henson P. M. The immunologic release of constituents from neutrophil leukocytes. II. Mechanisms of release during phagocytosis, and adherence to nonphagocytosable surfaces. J Immunol. 1971 Dec;107(6):1547–1557. [PubMed] [Google Scholar]
  19. Janoff A. Neutrophil proteases in inflammation. Annu Rev Med. 1972;23:177–190. doi: 10.1146/annurev.me.23.020172.001141. [DOI] [PubMed] [Google Scholar]
  20. Jensen M. S., Bainton D. F. Temporal changes in pH within the phagocytic vacuole of the polymorphonuclear neutrophilic leukocyte. J Cell Biol. 1973 Feb;56(2):379–388. doi: 10.1083/jcb.56.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Karnovsky M. L. The metabolism of leukocytes. Semin Hematol. 1968 Apr;5(2):156–165. [PubMed] [Google Scholar]
  22. Klebanoff S. J. Intraleukocytic microbicidal defects. Annu Rev Med. 1971;22:39–62. doi: 10.1146/annurev.me.22.020171.000351. [DOI] [PubMed] [Google Scholar]
  23. MOVAT H. Z., URIUHARA T., MACMORINE D. L., BURKE J. S. A PERMEABILITY FACTOR RELEASED FROM LEUKOCYTES AFTER PHAGOCYTOSIS OF IMMUNE COMPLEXES AND ITS POSSIBLE ROLE IN THE ARTHUS REACTION. Life Sci. 1964 Sep;3:1025–1032. doi: 10.1016/0024-3205(64)90115-8. [DOI] [PubMed] [Google Scholar]
  24. Malawista S. E., Bodel P. T. The dissociation by colchicine of phagocytosis from increased oxygen consumption in human leukocytes. J Clin Invest. 1967 May;46(5):786–796. doi: 10.1172/JCI105579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nachman R., Hirsch J. G., Baggiolini M. Studies on isolated membranes of azurophil and specific granules from rabbit polymorphonuclear leukocytes. J Cell Biol. 1972 Jul;54(1):133–140. doi: 10.1083/jcb.54.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schlesinger M. J., Reynolds J. A., Schlesinger S. Formation and localization of the alkaline phosphatase of Escherichia coli. Ann N Y Acad Sci. 1969 Oct 14;166(2):368–379. doi: 10.1111/j.1749-6632.1969.tb46408.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Stossel T. P., Pollard T. D., Mason R. J., Vaughan M. Isolation and properties of phagocytic vesicles from polymorphonuclear leukocytes. J Clin Invest. 1971 Aug;50(8):1745–1747. doi: 10.1172/JCI106664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Wetzel B. K., Horn R. G., Spicer S. S. Fine structural studies on the development of heterophil, eosinophil, and basophil granulocytes in rabbits. Lab Invest. 1967 Mar;16(3):349–382. [PubMed] [Google Scholar]
  31. Wetzel B. K., Spicer S. S., Dvorak H. F., Heppel L. A. Cytochemical localization of certain phosphatases in Escherichia coli. J Bacteriol. 1970 Oct;104(1):529–542. doi: 10.1128/jb.104.1.529-542.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wetzel B. K., Spicer S. S., Horn R. G. Fine structural localization of acid and alkaline phosphatases in cells of rabbit blood and bone marrow. J Histochem Cytochem. 1967 Jun;15(6):311–334. doi: 10.1177/15.6.311. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. ZUCKER-FRANKLIN D., HIRSCH J. G. ELECTRON MICROSCOPE STUDIES ON THE DEGRANULATION OF RABBIT PERITONEAL LEUKOCYTES DURING PHAGOCYTOSIS. J Exp Med. 1964 Oct 1;120:569–576. doi: 10.1084/jem.120.4.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zucker-Franklin D., Elsbach P., Simon E. J. The effect of the morphine analog levorphanol on phagocytosing leukocytes. A morphologic study. Lab Invest. 1971 Nov;25(5):415–421. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES