Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1969 Jan 31;129(2):267–293. doi: 10.1084/jem.129.2.267

LYSOSOMAL AND ULTRASTRUCTURAL CHANGES IN HUMAN "TOXIC" NEUTROPHILS DURING BACTERIAL INFECTION

Charles E McCall 1, Isao Katayama 1, Ramzi S Cotran 1, Maxwell Finland 1
PMCID: PMC2138608  PMID: 5762047

Abstract

"Toxic" neutrophils from humans with severe bacterial infections, identified by the presence of Döhle bodies, "toxic" granules, and vacuoles were shown to differ from normal neutrophils both in ultrastructure and in lysosome activity. Döhle bodies were identified as lamellar aggregates of rough endoplasmic reticulum. Toxic granules corresponded to the azurophilic granules usually identified by Romanowsky stains only in neutrophil precursors. By electron microscopy such granules were large, electron-dense, and peroxidase positive; they could usually be distinguished from the smaller, less dense, "specific" granules also present in control neutrophils, but in the latter they became visible by light microscopy only after prolonged staining or following fixation with glutaraldehyde. These observations suggest that toxic granules represent an abnormal staining reaction of the large dense granules in the toxic cells, and not phagocytized material, newly formed abnormal granules or autophagic bodies. Alkaline phosphatase activity was significantly greater in toxic neutrophils than in normal ones; 80% of the activity of both was located in the lysosome fraction. Beta glucuronidase was normal. Total acid phosphatase was normal, but the percentage located in the nonlysosome fraction of toxic neutrophils was increased, suggesting that lysosomes were "labilized." Formation of neutral red vacuoles in supravitally stained preparations, an index of lysosome activity, occurred more rapidly in toxic neutrophils. This reaction paralleled degranulation and the formation of clear vacuoles in unstained wet mounts and could be blocked by colchicine, a lysosome stabilizer, or enhanced by procedures which activate lysosomes. "Autophagic" vacuoles were observed by electron microscopy in some toxic neutrophils. These observations are discussed in relation to the concept that the "toxic" neutrophils in severe bacterial infection reflect cellular immaturity and/or stimulation or degeneration.

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

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

  1. ACKERMAN G. A. HISTOCHEMICAL DIFFERENTIATION DURING NEUTROPHIL DEVELOPMENT AND MATURATION. Ann N Y Acad Sci. 1964 Feb 28;113:537–565. doi: 10.1111/j.1749-6632.1964.tb40690.x. [DOI] [PubMed] [Google Scholar]
  2. ALLISON A. C., MALLUCCI L. HISTOCHEMICAL STUDIES OF LYSOSOMES AND LYSOSOMAL ENZYMES IN VIRUS-INFECTED CELL CULTURES. J Exp Med. 1965 Mar 1;121:463–476. doi: 10.1084/jem.121.3.463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. ALLISON A. C., YOUNG M. R. UPTAKE OF DYES AND DRUGS BY LIVING CELLS IN CULTURE. Life Sci. 1964 Dec;3:1407–1414. doi: 10.1016/0024-3205(64)90082-7. [DOI] [PubMed] [Google Scholar]
  4. Abernathy M. R. Döhle bodies associated with uncomplicated pregnancy. Blood. 1966 Mar;27(3):380–385. [PubMed] [Google Scholar]
  5. Anderson D. R. Ultrastructure of normal and leukemic leukocytes in human peripheral blood. J Ultrastruct Res. 1966 Sep;9:1–42. doi: 10.1016/s0022-5320(66)90002-5. [DOI] [PubMed] [Google Scholar]
  6. BESSIS M., THIERY J. P. Electron microscopy of human white blood cells and their stem cells. Int Rev Cytol. 1961;12:199–241. doi: 10.1016/s0074-7696(08)60541-0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Breton-Gorius J. Structures périodiques dans les granulations éosinophiles et neutrophiles des leucocytes polynucléaires du sang de l'homme. Nouv Rev Fr Hematol. 1966 Mar-Apr;6(2):195–208. [PubMed] [Google Scholar]
  9. Brittinger G., Hirschhorn R., Douglas S. D., Weissmann G. Studies on lysosomes. XI. Characterization of a hydrolase-rich fraction from human lymphocytes. J Cell Biol. 1968 May;37(2):394–411. doi: 10.1083/jcb.37.2.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Cohn Z. A., Hirsch J. G., Fedorko M. E. The in vitro differentiation of mononuclear phagocytes. IV. The ultrastructure of macrophage differentiation in the peritoneal cavity and in culture. J Exp Med. 1966 Apr 1;123(4):747–756. doi: 10.1084/jem.123.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Enomoto T., Kitani T. [Electron microscopic studies on peroxidase and acid phosphatase reaction in human leukocytes (in normal and leukemic cells and on phagocytosis)]. Nihon Ketsueki Gakkai Zasshi. 1966 Aug;29(4):554–570. [PubMed] [Google Scholar]
  16. FALLON H. J., FREI E., 3rd, DAVIDSON J. D., TRIER J. S., BURK D. Leukocyte preparations from human blood: evaluation of their morphologic and metabolic state. J Lab Clin Med. 1962 May;59:779–791. [PubMed] [Google Scholar]
  17. FOLLETTE J. H., VALENTINE W. N., LAWRENCE J. S. The beta glucuronidase content of human leukocytes in health and in disease. J Lab Clin Med. 1952 Dec;40(6):825–840. [PubMed] [Google Scholar]
  18. Fedorko M. E., Hirsch J. G. Cytoplasmic granule formation in myelocytes. An electron microscope radioautographic study on the mechanism of formation of cytoplasmic granules in rabbit heterophilic myelocytes. J Cell Biol. 1966 May;29(2):307–316. doi: 10.1083/jcb.29.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fedorko M. Effect of chloroquine on morphology of cytoplasmic granules in maturing human leukocytes--an ultrastructural study. J Clin Invest. 1967 Dec;46(12):1932–1942. doi: 10.1172/JCI105683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Herion J. C., Spitznagel J. K., Walker R. I., Zeya H. I. Pyrogenicity of granulocyte lysosomes. Am J Physiol. 1966 Sep;211(3):693–698. doi: 10.1152/ajplegacy.1966.211.3.693. [DOI] [PubMed] [Google Scholar]
  22. Hirschhorn R., Brittinger G., Hirschhorn K., Weissmann G. Studies on lysosomes. XII. Redistribution of acid hydrolases in human lymphocytes stimulated by phytohemagglutinin. J Cell Biol. 1968 May;37(2):412–423. doi: 10.1083/jcb.37.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. ITOGA T., LASZLO J. Dohle bodies and other granulocytic alterations during chemotherapy with cyclophosphamide. Blood. 1962 Dec;20:668–674. [PubMed] [Google Scholar]
  24. JANOFF A., WEISSMANN G., ZWEIOFACH B. W., THOMAS L. Pathogenesis of experimental shock. IV. Studies on lysosomes in normal and tolerant animals subjected to lethal trauma and endotoxemia. J Exp Med. 1962 Oct 1;116:451–466. doi: 10.1084/jem.116.4.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. JANOFF A., ZWEIFACH B. W. PRODUCTION OF INFLAMMATORY CHANGES IN THE MICROCIRCULATION BY CATIONIC PROTEINS EXTRACTED FROM LYSOSOMES. J Exp Med. 1964 Nov 1;120:747–764. doi: 10.1084/jem.120.5.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. JORDAN S. W., LARSEN W. E. ULTRASTRUCTURAL STUDIES OF THE MAY-HEGGLIN ANOMALY. Blood. 1965 Jun;25:921–932. [PubMed] [Google Scholar]
  27. Koszewski B. J., Vahabzadeh H., Willrodt S. Hemosiderin content of leukocytes in animals and man and its significance in the physiology of granulocytes. Am J Clin Pathol. 1967 Nov;48(5):474–483. doi: 10.1093/ajcp/48.5.474. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. OSKI F. A., NAIMAN J. L., ALLEN D. M., DIAMOND L. K. Leukocytic inclusions--Dohle bodies--associated with platelet abnormality (the May-Hegglin anomaly). Report of a family and review of the literature. Blood. 1962 Dec;20:657–667. [PubMed] [Google Scholar]
  30. PALADE G. E. A study of fixation for electron microscopy. J Exp Med. 1952 Mar;95(3):285–298. doi: 10.1084/jem.95.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. THOMAS L. POSSIBLE ROLE OF LEUCOCYTE GRANULES IN THE SHWARTZMAN AND ARTHUS REACTIONS. Proc Soc Exp Biol Med. 1964 Jan;115:235–240. doi: 10.3181/00379727-115-28879. [DOI] [PubMed] [Google Scholar]
  32. Trump B. F., Bulger R. E. New ultrastructural characteristics of cells fixed in a glutaraldehyde-osmium tetroxide mixture. Lab Invest. 1966 Jan;15(1 Pt 2):368–379. [PubMed] [Google Scholar]
  33. Watanabe I., Donahue S., Hoggatt N. Method for electron microscopic studies of circulating human leukocytes and observations on their fine structure. J Ultrastruct Res. 1967 Oct 31;20(5):366–382. doi: 10.1016/s0022-5320(67)80106-0. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. White J. G. The Chediak-Higashi syndrome: a possible lysosomal disease. Blood. 1966 Aug;28(2):143–156. [PubMed] [Google Scholar]
  36. Yamada E. Electron microscopy of the peroxidase in the granular leucocytes of rat bone marrow. Arch Histol Jpn. 1966 Nov;27(1):131–138. doi: 10.1679/aohc1950.27.131. [DOI] [PubMed] [Google Scholar]
  37. Zieve P. D., Haghshenass M., Blanks M., Krevans J. R. Vacuolization of the neutrophil. An aid in the diagnosis of septicemia. Arch Intern Med. 1966 Oct;118(4):356–357. [PubMed] [Google Scholar]
  38. Zucker-Franklin D. The phagosomes in rheumatoid synovial fluid leukocytes: a light, fluorescence, and electron microscope study. Arthritis Rheum. 1966 Feb;9(1):24–36. doi: 10.1002/art.1780090104. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES