Summary
Using a scanning microscope photometer we determined quantitatively the enzymecytochemical reaction product for naphthol-AS-D-chloroacetate esterase in neutrophilic granulocytes and their precursors in man.
Evaluation of neutrophilic cells from three healthy donors resulted in a logarithm-normal distribution. After subdivision of these cells in their morphologically defined maturational stages no statistically bimodal distribution was shown within the single cell groups. Myelocytes showed twice the amount of the polymorphonuclear neutrophil absorption values. The highest promyelocyte obsorptions were double the values of the myelocyte absorptions. The standard deviation of the absorbance obtained with promyelocytes (which encompass cells already producing granules up to cells reaching their maximal granule content) was significantly higher than the standard deviation of the myelocytes. As already known, primary granules are only synthesized at the promyelocyte stage and — according to the present knowledge — their chloracylesterase and peroxidase activities are not lost during further maturation. Consequently, our results indicate that only enzyme-rich, late promyelocytes undergo mitosis transforming into myelocytes. Correspondingly, their absorption value was halved. Since the absorbance from myelocytes to polymorphonuclears is again halved, myelocytes divide only once. Metamyelocyte absorptions in part correspond to that of myelocytes. This indicates that no distinction can be made between myelocytes with mitotic capacity and “true” if only the size and the nuclear shape are considered metamyelocytes which are not longer capable of undergoing mitosis.
Similar content being viewed by others
References
Bainton DF (1975) Neutrophil granules. Br J Haematol 29: 17–22
Bainton DF, Farquhar MG (1966) Origin of granules in polymorphonuclear leukocytes. Two types derived from opposite faces of the Golgi complex in developing granulocytes. J Cell Biol 28: 277–301
Bainton DF, Ullyot JL, Farquhar MG (1971) The development of neutrophilic polymorphonuclear leukocytes in human bone marrow. Origin and content of a urophil and specific granules. J Exp Med 134: 907–934
Blumenson LE (1973) A comprehensive modeling procedure for the human granulopoietic system: overall view and summary of data. Blood 42: 303–313
Boll I (1978) Das granulozytäre System. In: Queisser W (Hrsg) Das Knochenmark, Morphologie, Funktion, Diagnostik. Thieme, Stuttgart, S 167
Boll I, Kuhn A (1965) Granulocytopoiesis in human bone marrow cultures studied by means of kinematography. Blood 26: 449–470
Boll ITM, Sterry K, Maurer HR (1979) Evidence for a rat granulocyte chalone effect on the proliferation of normal human bone marrow and of myeloid leukemias. Acta Haematol 61: 130–137
Brederoo P, Daems WT (1978) The ultrastructure of Guinea pig heterophil granulocytes and the heterogeneity of the granules. I. Development in the bone marrow. Cell Tissue Res 194: 183–205
Bretz U, Baggiolini M (1974) Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J Cell Biol 63: 251–269
Dresch C, Flandrin G, Breton-Gorius J (1980) Phagocytosis of neutrophilic polymorphonuclears by macrophages in human bone marrow: inportance in granulopoiesis. J Clin Pathol 33: 1110–1113
Geissler G (in preparation) Anfärbungskinetik der Naphthol-AS-D-Chloroazetat Esterase. Med Diss, Köln
Hoffstein S, Soberman R, Goldstein I, Wissmann G (1976) Concanavalin A induces microtubule assembly and specific granule discharge in human polymorphinuclear leukocytes. J Cell Biol 68: 781–787
Killmann SA, Cronkite EP, Fliedner TM, Bond VP, Becher G (1963) Mitotic indices of human bone marrow cells. II. The use of mitotic indices for estimation or time parameters of proliferation in sereally connected multiplicative cellular compartments. Blood 21: 141–163
Klebanoff SJ, Clark RA (1978) The neutrophil: function and clinical disorders. North Holland, Amsterdam New York Oxford
Koeffler HP, Golde DW (1980) Human myeloid leukemia cell lines: a review. Blood 56: 344–350
Maalem H, Sheppard K, Fletcher J (1982) The discharge of primary and secondary granules during immune phagocytosis by normal and chronic granulocytic leukemia polymorphonuclear neutrophils. Br J Haematol 51: 201–208
Mackey MC, Dörmer P (1981) Enigmatic hemopoiesis. In: Rotenberg (ed) Biomathematics and cell kinetics. Elsevier-North Holland, Amsterdam, p 87
Mackey MC, Dörmer P (1982) Continuous maturation of proliferating erythroid precursors. Cell Tissue Kinet 15: 381–392
Mansberg HP, Saunders AM, Groner W (1974) The Hemalog D white cell differential system. J Histochem Cytochem 22: 711–724
Maurer AM (1965) Diurnal variation of proliferative activity in the human bone marrow. Blood 26: 1–7
Moloney WC, McPherson K, Fliegelman L (1960) Esterase activity in leukocytes demonstrated by the use of naphthol-AS-D-chloroacetate substrate. J Histochem Cytochem 8: 200–207
Olsson I, Olofsson T (1981) Induction of differentiation in a human promyelocytic leukemia cell line (HL 60). Production of granula proteins. Exp Cell Res 131: 225–230
Queisser W (1973) Die Anwendung der kombinierten cytophotometrisch-autoradiographischen Methodik für die Untersuchung der Proliferation normaler und gestörter hämopoietischer Zellsysteme. Klin Wochenschr 51: 687–694
Rindler R, Hortnagel H, Schmalzl F, Braunsteiner H (1973) Hydrolysis of a chymotrypsin substrate and of naphthol-AS-D-chloroacetate by human leukocyte granules. Blut 26: 239–249
Rindler-Ludwig R, Braunsteiner H (1977) Biochemie der Granula myeloischer Zellen unter physiologischen und pathophysiologischen Bedingungen. Dtsch Med Wochenschr 102: 24–27
Rondanelli EG, Magliulo E, Giraldi A, Carco FP (1967) The chronology of the mitotic cycle of human granulopoietic cells. Phase contrast studies on living cells in vitro. Blood 30: 557–568
Schmalzl F, Wiedermann CJ (1983) Zur Technik quantitativer enzymzytochemischer Untersuchungen an polymorphkernigen Neutrophilen des Menschen. Acta Histochem [Suppl] 28: 157–162
Schmalzl F, Huhn D, Asamer H, Rindler R, Braunsteiner H (1973) Cytochemistry and ultrastructure of pathologic granulation in myelogenous leukemia. Blut 27: 243–252
Schultze B (1969) Autoradiography at the cellular level. In: Pollister AW (ed) Physical techniques in biological research, vol III: B, 2nd edn. Academic Press, New York, p 82
Smeby W, Benestad HB (1980) Simulation of murine granulopoiesis. Blut 41: 47–60
Spitznagel JK, Dalldorf FG, Leffels MS, Folds JD, Welsh IRH, Cooney MH, Martin LE (1974) Character of azurophil and specific granules purified from human polymorphonuclear leukocytes. Lab Invest 30: 774–785
Warner HR, Athens SW (1964) An analysis of granulocyte kinetics in blood and bone marrow. Ann NY Acad Sci 113: 523–532
Wichmann HE, Gross R (1981) How mathematical models can interpret and predict experimental results in hematology. Klin Wochenschr 59: 1–4
Wintrobe MW (1981) Clinical hematology, 8th edn. Lea and Fibinger, Philadelphia
Wright DG, Kauffmann JC, Terpstra GK, Graw RG, Deisseroth AB, Gallin Ji (1978) Mobilisation and exocytosis of specific (secondary) granules by human neutrophils during adherence to nylon wool in filtration leukapheresis (FL). Blood 52: 770–782
Author information
Authors and Affiliations
Additional information
Supported by the “Österreichische Nationalbank” and by grant No. 3759 of the “Fonds zur Förderung der wissenschaftlichen Forschung”
Rights and permissions
About this article
Cite this article
Wiedermann, C.J., Schmalzl, F. & Braunsteiner, H. Investigation of granulocytopoietic kinetics by microdensitometric evaluation of primary granule naphthol-AS-D-chloroacetate esterase activity. Blut 47, 271–277 (1983). https://doi.org/10.1007/BF00319896
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00319896