Summary
Unique fusiform or spindle-shaped particles (Phi bodies) and rods with hydroperoxidase (catalase and/or peroxidase) activity are present in human granulocyte precursors only in acute myelogenous leukemia (AML). These newly recognized particles are much more numerous and prominent than Auer rods. They may be rapidly and readily identified using the microscope in marrow or peripheral blood films when the procedures recommended in this paper for fixation, incubation for hydroperoxidase demonstration in 3,3′-diaminobenzidine (DAB)/H2O2 medium, copper salt treatment and counterstaining (optional) with the Papanicolaou method are employed. Films prepared in the same manner but treated with benzidine/H2O2 medium for myeloperoxidase did not reveal these particles.
We believe that Phi bodies are pathognomonic of AML since they are almost invariably present in AML patients with active disease. Their presence serves to distinguish AML from acute lymphocytic leukemia and from chronic granulocytic leukemia in blast crisis. Since the particles disappear in disease remission and reappear upon relapse, the recommended procedure is not only useful in diagnosis but in guiding therapy.
When a very rapid diagnosis is needed, it is not necessary to counterstain the preparations, but the nuclei, cytoplasm and plasmalemma can readily be observed in the granulocyte precursors when they are counterstained by the Papanicolaou method. This treatment does not diminish the clarity of the Phi bodies and rods which stain by virtue of their peroxidatic activity.
This cytochemical diagnostic procedure should be considered for adoption by hematology laboratories.
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Breton-Gorius, J., Coquin, Y., Guichard, J.: Cytochemical distinction between azurophils and catalase-containing granules in leukocytes. Lab. Invest 38, 21–31 (1978)
Gill, G.W., Frost, J.K., Miller, K.A.: A new formula for a half-oxidized hematoxylin solution that neither overstains nor requires differentiation. Acta Cytol. 18, 300–311 (1974)
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. 14, 291–301 (1966a)
Graham, R.C., Jr., Karnovsky, M.J.: Glomerular permeability. Ultrastructural cytochemical studies using peroxidases as protein tracers. J. Exp. Med 124, 1123–1133 (1966b)
Hanker, J.S., Romanovicz, D.K.: Phi bodies: peroxidatic particles that produce crystalloidal cellular inclusions. Science 197, 895–898 (1977)
Hanker, J.S., Moore, J.O., Ambrose, W.W., James, C.J., Mandelkorn, J., Yates, P.E., Gall, S.A., Bossen, E.H., Fay, J.W., Laszlo, J.: Facilitated light microscopic cytochemical diagnosis of acute myeloid leukemia. Cancer Research (Submitted for publication, 1979)
Humason, G.L.: Animal tissue techniques, Third Edition. San Francisco: W.H. Freeman and Company 1972
Ichimaru, M.: Studies on leukocyte catalase especially in leukemic cells. J. Kyushu Hematol. Soc. 9, 722–753 (1959)
Kaplow, L.S.: Simplified myeloperoxidase stain using benzidine dihydrochloride. Blood 26, 215 (1965)
Kaplow, L.S.: Substitutes for benzidine in myeloperoxidase stains. Am. J. Clin. Pathol. 63, 451 (1975)
Kidson, C.: Variations in catalase activity in human leukocytes. Blood 19, 82–88 (1962)
Luna, L.G.: Manual of histologic staining methods of the Armed Forces Institute of Pathology. Third Edition. New York: McGraw-Hill 1968
Moore, J.O., Laszlo, J., Hanker, J.S.: In preparation (1979)
Nichols, T.M.: An electron microscopic investigation of Auer bodies in myeloblastic leukemia. C.M.A. J. 102, 865–867 (1970)
Nishimura, E.T., Cooper, C.: Peroxidatic reaction of catalase-antibody complex of leukocytes demonstrated by diaminobenzidine. Cancer Res. 34, 2386–2392 (1974)
Nishimura, E.T., Whest, G.M., Yang, H-Y.: Ultrastructural localization of peroxidatic catalase in human peripheral blood leukocytes. Lab. Invest. 34, 60–68 (1976)
Novikoff, A.B. and Goldfischer, S.: Visualization of peroxisomes (microbodies) and mitochondria with diaminobenzidine. J. Histochem. Cytochem. 17, 675–680 (1969)
Rausch, P.G.: Personal Communication (1978)
Roels, F., Wisse, E., DePrest, B., van der Meulen, J.: Cytochemical discrimination between catalases and peroxidases using diaminobenzidine. Histochemistry 41, 281–312 (1975)
Szmigielski, S.: Peroxisomal enzymes in human granulocytes. I. Catalase and peroxidase. Folia Histochem. Cytochem. 10, 47–50 (1972)
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This investigation was supported by NIH research grants DE02668, CA11265, DE04730, and RR05333
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Hanker, J.S., Laszlo, J. & Moore, J.O. The light microscopic demonstration of hydroperoxidase-positive Phi bodies and rods in leukocytes in acute myeloid leukemia. Histochemistry 58, 241–252 (1978). https://doi.org/10.1007/BF00495380
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DOI: https://doi.org/10.1007/BF00495380