Skip to main content
Log in

Enzyme cytochemistry of blood and marrow cells

  • Review
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Synopsis

Diseases of the blood and bone marrow are commonly associated with abnormalities of oxido-reductase and lysosomal enzymes within individual erythrocytes and leucocytes. There are considerable technical difficulties, however, in adapting enzyme histochemical techniques to the study of haemopoietic tissue since individual cells are readily disrupted during processing, show variable enzyme activity according to the stage of maturation, and possess a lipoprotein cytoplasmic membrane which hinders reagent penetration.

Cytochemical techniques for the study of oxido-reductase systems are of importance in the study of the neutrophil in infected patients, the erythrocyte in glucose-6-phosphate dehydrogenase deficiency, and the primitive blast cell in acute leukaemia. Lysosomal enzymes are of importance in the study of the neutrophil in infected patients and in the differential diagnosis of acute leukaemia. Some examples of recent studies of these enzyme systems are given to illustrate technical procedures involving cytocentrifugation of cells on to glass slides, adjustment of the osmolality of the reaction mixture, and the study of smeared cells as opposed to cells incubated in suspension.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ackerman, G. A. (1960). Histochemical demonstration of dehydrogenase activity in the cells of normal human blood and bone marrow.J. biophys. biochem. Cytol. 8, 61–7.

    Google Scholar 

  • Ackerman, G. A. (1962). Substitured naphthol AS phosphate derivatives for the localization of leukocyte alkaline phosphatase activity.Lab. Invest. II, 563–7.

    Google Scholar 

  • Allison, A. C. &Mallucci, L. (1964). Lysosomes in dividing cells, with special reference to lymphocytes.Lancet 2, 1371–3.

    Google Scholar 

  • Asamer, H., Schmalzl, F. &Braunsteiner, H. (1969). Der immunzytologische lysozymnachweis in menschlichen blutzellen.Acta haemat. (Basel) 41, 49–54.

    Google Scholar 

  • Asamer, H., Schmalzl, F. &Braunsteiner, H. (1971). Immunocytological demonstration of lysozyme (muramidase) in human leukaemic cells.Brit. J. Haemat. 20, 571–4.

    Google Scholar 

  • Baehner, R. L. &Karnovsky, M. L. (1968). Deficiency of reduced nicotinamide-adenine dinucleotide oxidase in chronic granulomatous disease.Science 162, 1277–9.

    Google Scholar 

  • Baehner, R. L. &Nathan, D. G. (1967). Leukocyte oxidase: defective activity in chronic granulomatous disease.Science 155, 835–6.

    Google Scholar 

  • Baehner, R. L. &Nathan, D. G. (1968). Quantitative nitroblue tetrazolium test in chronic granulomatous disease.New Engl. J. Mod. 278, 971–6.

    Google Scholar 

  • Bainton, D. F. &Farquhar, M. G. (1966). Origin of granules in polymorphonuclear leukocytes.J. Cell Biol. 28, 277–301.

    Google Scholar 

  • Bainton, D. F. &Ullyot, J. L. (1974). Azurophil and specific granules of blood neutrophils in chronic myelogenous leukaemia: an ultrastructural and cytochemical analysis.Brit. J. Haemat. 28, 149.

    Google Scholar 

  • Bernstein, R. E. (1962). A rapid screening dye test for the detection of glucose-6-phosphate dehydrogenase deficiency in red cells.Nature (Lond.) 194, 192–3.

    Google Scholar 

  • Bitensky, L., Butcher, R. G., Johnstone, J. J. &Chayen, J. (1974). Effect of glucocorticoids on lysosomes in synoviallining cells in human rheumatoid arthritis.Ann. rheum. Dis. 33, 57–61.

    Google Scholar 

  • Bittner, S. J., Kieff, E., Windhorst, D. &Meier, P. (1973). The use of the unstimulated nitroblue tetrazolium test as a routine screening test for bacterial infection in an adult population.Am. J. clin. Path. 60, 843–53.

    Google Scholar 

  • Björkstén, B. (1973). The NBT test using venous and capillary blood.Scand. J. Haemat. 11, 270–4.

    Google Scholar 

  • Bohn, L. (1965). Histochemical demonstration of lactate dehydrogenase in cells in blood films.Scand. J. Haemat. 2, 108–17.

    Google Scholar 

  • Boysen, G. (1969). An evaluation of aryl sulphatase activity in leukaemic cells.Scand. J. Haemat. 6, 246–9.

    Google Scholar 

  • Böyum, A. (1968). Separation of leucocytes from blood and bone marrow.Scand. J. clin. Lab. Invest. 21, Suppl. 97, 1–109.

    Google Scholar 

  • Boxer, G. E. (1965). Glycolytic enzymes in human tumors: difficulties, methods and results.Eur. J. Cancer 1, 161–72.

    Google Scholar 

  • Briggs, R. S., Perillie, P. E. &Finch, S. C. (1966). Lysozyme in bone marrow and peripheral blood cells.J. Histochem. Cytochem. 14, 167–70.

    Google Scholar 

  • Gaspersson, T. &Lomakka, G. (1962). Scanning microscopy techniques for high resolution quantitative cytochemistry.Ann. N. Y. Acad. Sci. 97, 449–63.

    Google Scholar 

  • Catovsky, D. &Galton, D. A. G. (1973). Lysozyme activity and nitroblue-tetrazolium reduction in leukaemic cells.J. clin. Path. 26, 60–9.

    Google Scholar 

  • Catovsky, D., Galton, D. A. G. &Robinson, J. (1972). Myeloperoxidase-deficient neutrophils in acute myeloid leukaemia.Scand. J. Haemat. 9, 142–8.

    Google Scholar 

  • Catovsky, D., Galetto, J., Okos, A., Miliani, E. &Galton, D. A. G. (1975). Cytochemical profiles of B and T leukaemic lymphocytes.J. clin. Path. 27, 767–71.

    Google Scholar 

  • Catovsky, D., Pettit, J. E., Okos, A. &Galton, D. A. G. (1974). The B-lymphocyte nature of the hairy cell of leukaemic reticuloendotheliosis.Brit. J. Haemat. 26, 29–37.

    Google Scholar 

  • Chayen, J., Bitensky, L. &Butcher, R. G. (1973).Practical Histochemistry. London: Wiley.

    Google Scholar 

  • Cohn, Z. A. &Hirsch, J. G. (1960). The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leukocytes.J. exp. Med. 112, 983–1004.

    Google Scholar 

  • Deeley, E. M. (1955). An integrating microdensitometer for biological cells.J. scient. Instrum. 32, 263–7.

    Google Scholar 

  • Djerassi, I., Kim, J. S., Suvansri, U., Mitrakul, C. &Cieselka, W. (1972). Continuous flow filtration-leukopheresis.Transfusion 12, 75–83.

    Google Scholar 

  • Doré, C. F. &Balfour, B. M. (1965). A device for preparing cell spreads.Immunology,9, 403–5.

    Google Scholar 

  • Douglas, S. D., Cohnen, G., König, E. &Brittinger, G. (1973). Lymphocyte lysosomes and lysosomal enzymes in chronic lymphocytic leukermi.Blood 41, 511–18.

    Google Scholar 

  • Edelson, P. J., Stites, D. P., Gold, S. &Fudenberg, H. H. (1973). Disorders of neutrophil function: defects in the early stages of the phagocytic process.Clin. exp. Immunol. 13, 21–8.

    Google Scholar 

  • Evans, W. H., Wolf, M. M. &Chabner, B. A. (1974). Concentration of immature and mature granulocytes from normal human bone marrow.Proc. Soc. exp. Biol. (N. Y.) 146, 526–9.

    Google Scholar 

  • Fairbanks, V. F. &Fernandez, M. N. (1969). The identification of metabolic errors associated with hemolytic anemia.J. Am. med. Assoc. 208, 316–20.

    Google Scholar 

  • Fairbanks, V. F. &Lampe, L. T. (1968). A tetrazolium-linked cytochemical method for estimation of glucose-6-phosphate dehydrogenase activity in individual erythrocytes: applications in the study of heterozygotes for glucose-6-phosphate dehydrogenase deficiency.Blood 31, 589–603.

    Google Scholar 

  • Flandrin, G. &Daniel, M. T. (1973). Practical value of cytochemical studies for the classification of acute leukemias. In:Nomenclature, Methodology and Results of Clinical Trials in Acute Leukemias (eds. G. Mathé, P. Pouillart & P. Pohwarzenbergg) pp. 43–56. London: Heinemann.

    Google Scholar 

  • Gahrton, G. (1965). A quantified periodic acid-Schiff reaction in normal and leukemic leukocytes.Eur. J. Cancer 1, 299–302.

    Google Scholar 

  • Gall, J. C., Jr., Brewer, G. J. &Dern, R. J. (1965). Studies of glucose-6-phosphate dehydrogenase activity of individual erythrocytes: the methemoglobin-elution test for identification of females heterozygous for G6PD deficiency.Am. J. hum. Genet. 17, 359–68.

    Google Scholar 

  • Gifford, R. H. &Malawista, S. E. (1970). A simple rapid micromethod for detecting chronic granulomatous disease of childhood.J. Lab. clin. Med. 75, 511–19.

    Google Scholar 

  • Glick, A. D. &Horn, R. G. (1974). Identification of promonocytes and monocytoid precursors in acute leukaemia of adults: ultrastructural and cytochemical observations.Brit. J. Haemat. 26, 395–403.

    Google Scholar 

  • Goldberg, A. F. &Barka, T. (1962). Acid phosphatase activity in human blood cells.Nature 195, 297.

    Google Scholar 

  • Gordon, A. M., Rowan, R. M., Brown, T. &Carson, H. G. (1973). Routine application of the nitroblue tetrazolium test in the clinical laboratory.J. clin. Path. 26, 52–6.

    Google Scholar 

  • Gordon, A. M.,&Stuart, J. (1974). Red cell cytochemistry in G-6-PD deficiency.Brit. J. Haemat. 27, 358.

    Google Scholar 

  • Gordon, P. A., Stuart, J., Lee, T. R., Breeze, G. R. &Pugh, R. N. H. (1975). The cytocentrifuge NBT test.J. clin. Path. 28, 674–9.

    Google Scholar 

  • Graw, R. G., Jr., Herzig, G. P., Eisel, R. J. &Perry, S. (1971). Leukocyte and platelet collection from normal donors with the continuous flow blood cell separator.Transfusion 11, 94–101.

    Google Scholar 

  • Harris, R. &Ukaejiofo, E. O. (1970). Tissue typing using a routine one-step lymphocyte separation procedure.Brit. J. Haemat. 18, 229–35.

    Google Scholar 

  • Hayashi, M., Nakajima, Y. &Fishman, W. H. (1964). The cytologic demonstration of β-glucuronidase employing naphthol AS-BI glucuronide and hexazonium pararosanilin; a preliminary report.J. Histochem. Cytochem. 12, 293–7.

    Google Scholar 

  • Hayhoe, F. G. J., Quaglino, D. &Doll, R. (1964).The Cytology and Cytochemistry of Acute Leukaemias London: H.M.S..O.

    Google Scholar 

  • Karle, H., Hansen, N. E. &Killmann, S-A. (1974). Intracellular lysozyme in mature neutrophils and blast cells in acute leukemia.Blood 44, 247–55.

    Google Scholar 

  • Keller, D. F. (1971).Glucose-6-phosphate Dehydrogenase Deficiency. C.R.C. Monoscience Series. London: Butterworths.

    Google Scholar 

  • Iake, B. D. (1966). The histochemistry of phosphatases: the use of lead acetate instead of lead nitrate.J. R. microsc. Soc. 85, 73–5.

    Google Scholar 

  • Lawrinson, W. &Gross, S. (1964). Nuclear arylsulfatase activity in primitive hemic cells.Lab. Invest. 13, 1612–6.

    Google Scholar 

  • Lee, T. R. & Stuart, J. (1975). Unpublished observation.

  • Leinonen, E. A. (1970). Cytochemical studies of acute leukemias.Acta haemat. (Basel) 43, 219–27.

    Google Scholar 

  • Libnoch, J. A., Yakulis, V. J. &Heller, P. (1966). Lactate dehydrogenase in megaloblastic bone marrow.Am. J. clin. Path. 45, 302–5.

    Google Scholar 

  • Lyon, M. (1961). Gene action in the X-chromosome of the mouse (Mus musculus L.).Nature (Lond.) 190, 372–3.

    Google Scholar 

  • Mann, J. R., Simpson, J. S., Munkley, R. M. &Stuart, J. (1971). Lysosomal enzyme cytochemistry in acute leukaemia.J. clin. Path. 24, 831–6.

    Google Scholar 

  • Martin, E. (1959). Increase of the cathepsin activity of the liver and of skeletal muscle of rats treated either with 2,4-dinitrophenol or with bacterial lipopolysacchirides.Experientia (Basel) 15, 182–3.

    Google Scholar 

  • Motulsky, A. G., Kraut, J. M., Thieme, W. T. &Musto, D. F. (1959). Biochemical genetics glucose-6-phosphate dehydrogenase deficiency.Clin. Res. 7, 89–90.

    Google Scholar 

  • Nakao, M., Nakayama, T. &Kankura, T. (1973). A new method for separation of human blood components.Nature, New Biol. 246, 94.

    Google Scholar 

  • Panizon, F., Cramer, R., Soranzo, M. R. &Zabucchi, G. (1974). Leucocyte disfunction in Chediak-Higashi syndrome.Abstracts, 4th meeting, European Society for Paediatric Haematology and Immunology, pp. 316–17. Berlin. Springer-Verlag.

    Google Scholar 

  • Park, B. H., Fikrig, S. M. &Smithwick, E. M. (1968). Infection and nitroblue tetrazolium reduction by neutrophils. A diagnostic aid.Lancet 2, 532–4.

    Google Scholar 

  • Park, B. H. &Good, R. A. (1970). N.B.T. test stimulated.Lancet 2, 616.

    Google Scholar 

  • Paul, J. (1966). Metabolic processes in normal and cancer cells. In:The Biology of Cancer (eds. E.J. Ambrose & F. J. C. Rose) pp. 52–64. London: Van Nostrand.

    Google Scholar 

  • Pearse, A. G. E. (1972).Histochemistry: Theoretical and Applied, 3rd Edn., p. 896. London: Churchill.

    Google Scholar 

  • Pedersen, B. &Hayhoe, F. G. J. (1971a). Cellular changes in chronic myeloid leukaemia.Brit. J. Haemat. 21, 251–6.

    Google Scholar 

  • Pedersen, B. &Hayhoe, F. G. J. (1971b). Relation between phagocytic activity and alkaline phosphatase content of neutrophils in chronic myeloid leukaemia.Brit. J. Haemat. 21, 257–60.

    Google Scholar 

  • Root, R. K., Rosenthal, A. S. &Balestra, D. J. (1972). Abnormal bactericidal, metabolic and lysosomal functions of Chediak-Higashi syndrome leukocytes.J. clin. Invest. 51, 649–65.

    Google Scholar 

  • Rosales, C. L., Bennett, J. M. &Rutenberg, A. M. (1965). An azo dye method for the histochemical demonstration of leukocyte acid phosphatase.J. Histochem. Cytochem. 13, 714.

    Google Scholar 

  • Rozenszajn, L., Leibovich, M., Shoham, D. &Epstein, J. (1968). The esterase activity in megaloblasts, leukaemic and normal haemopoietic cells.Brit. J. Haemat. 14, 605–10.

    Google Scholar 

  • Rozenszajn, L. &Shoham, D. (1967). The demonstration of dehydrogenases and diaphorases in cells of peripheral blood and bone marrow.Blood 29, 737–46.

    Google Scholar 

  • Rozenszajn, L. A., Shoham, D. &Menashi, T. (1972). Evaluation of glucose-6-phosphatase dehydrogenase in single erythrocytes in human blood smears.Acta haemat. (Basel) 47, 303–10.

    Google Scholar 

  • Schmalzl, F., Lederer, B. &Braunsteiner, H. (1970). Atypical myeloblastic leukaemia with differentiation into paraneutrophils.Blut 20, 337–49.

    Google Scholar 

  • Segal, A. W. &Levi, A. J. (1973). The mechanism of the entry of dye into neutrophils in the nitroblue tetrazolium (NBT) test.Clin. Sci. Mol. Med. 45, 817–26.

    Google Scholar 

  • Segal, A. W., Trustey, S. F. &Levi, A. J. (1973). Re-evaluation of the nitroblue tetrazolium test.Lancet 2, 879–83.

    Google Scholar 

  • Shehan, H. L. &Storey, G. W. (1947). An improved method of staining leucocyte granules with Sudan black B.J. Path. Bact. 59, 336–7.

    Google Scholar 

  • Smutka, P. &Brunning, R. D. (1969). An evaluation of nuclear aryl sulfatase activity in acute leukemias.Acta haemat. (Basel) 41, 290–5.

    Google Scholar 

  • Steigbigel, R. T., Johnson, P. K. &Remington, J. S. (1974). The nitroblue tetrazolium reduction test versus conventional hematology in the diagnosis of bacterial infection.New Engl. J. Med. 290, 235–8.

    Google Scholar 

  • Stuart, J., Gordon, P. A. &Lee, T. R. (1975). Polymorph cytochemistry in systemic infection.Brit. J. Haemat. 30, 126–7.

    Google Scholar 

  • Stuart, J. &Simpson, J. S. (1970). Dehydrogenase enzyme cytochemistry of unfixed leucocytes.J. clin. Path. 23, 517–21.

    Google Scholar 

  • Stuart, J., Simpson, J. S. &Mann, J. R. (1970). Intracellular hydrogen transport systems in acute leukaemia.Brit. J. Haemat. 19, 739–48.

    Google Scholar 

  • Stuart, J., Simpson, J. S. &Wallin, J. M. (1972). Glycolytic and respiratory enzyme activity in relation to leucocyte maturation.Brit. J. Haemat. 23, 79–87.

    Google Scholar 

  • Syrén, E. &Raeste, A. M. (1971). Identification of blood monocytes by demonstration of lysozyme and peroxidase activity.Acta haemat. (Basel) 45, 29–35.

    Google Scholar 

  • Tew, J. G., Scott, R. L. &Donaldson, D. M. (1971). Plasma β-lysin and lysozyme following endotoxin administration and the generalized Shwartzman reaction.Proc. Soc. exp. Biol. (N. Y.) 136, 473–8.

    Google Scholar 

  • Tzortzatou, F. &Hayhoe, F. G. J. (1974a). Cytochemical demonstration of dihydrofolate reductase activity.Brit. J. Haemat. 28, 199–208.

    Google Scholar 

  • Tzortzatou, F. &Hayhoe, F. G. J. (1974b). The effect of folate antagonists on dihydrofolate reductase activity demonstrated cytochemically.Brit. J. Haemat. 28, 209–16.

    Google Scholar 

  • Warburg, O. (1930).The Metabolism of Tumours (translated by F. Dickens) London: Constable.

    Google Scholar 

  • Warburg, O. (1956). On the origin of cancer cells.Science 123, 309–14.

    Google Scholar 

  • Watson, P. (1966). A slide centrifuge: an apparatus for concentrating cells in suspension onto a microscope slide.J. Lab. clin. Med. 68, 494–501.

    Google Scholar 

  • Weissmann, G. &Thomas, L. (1962). Studies on lysosomes. I. The effects of endotoxin, endotoxin tolerance, and cortisone on the release of acid hydrolases from a granular fraction of rabbit liver.J. exp. Med. 116, 433–50.

    Google Scholar 

  • W. H. O. (1967). Standardization of procedures for the study of glucose-6-phosphatase dehydrogenase. World Health Organization Technical Report Series no. 366. Geneva.

  • Wulff, H. R. &Sørensen, M. (1966). A new technique for the histochemical demonstration of phosphorylase in blood films.Acta haemat. (Basel) 35, 304–10.

    Google Scholar 

  • Yam, L. T., Li, C. Y. &Crosby, W. J. (1971). Cytochemical identification of monocytes and granulocytes.Am. J. clin. Path. 55, 283–90.

    Google Scholar 

  • Yam, L. T., Li, C. Y. &Lam, K. W. (1971). Tartrate-resistant acid phosphatase isoenzyme in the reticulum cells of leukemic reticuloendotheliosis.New Engl. J. Med. 284, 357–60.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stuart, J., Gordon, P.A. & Lee, T.R. Enzyme cytochemistry of blood and marrow cells. Histochem J 7, 471–487 (1975). https://doi.org/10.1007/BF01003884

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01003884

Keywords

Navigation