Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1993 Oct;46(10):903–907. doi: 10.1136/jcp.46.10.903

Immunological and ultrastructural studies in acute biphenotypic leukaemia.

V Shetty 1, A Chitale 1, E Matutes 1, V Buccheri 1, R Morilla 1, D Catovsky 1
PMCID: PMC501615  PMID: 8227405

Abstract

AIMS--To compare the sensitivity of the ultrastructural method to detect myeloperoxidase (MPO) with light microscopy and immunocytochemistry using an anti-MPO antibody; to examine the expression of lymphoid antigens in relation to MPO activity in blast cells from cases of biphenotypic leukaemia. METHODS--Blast cells from 14 cases of biphenotypic acute leukaemia were analysed. Immunological markers were performed by single or double immunofluorescence staining on a flow cytometer. The presence of MPO was determined by light microscopy, electron microscopy on fixed and unfixed cells, and by immunoalkaline phosphatase with an anti-MPO antibody. The immunogold method was applied at the ultrastructural level to assess the expression of lymphoid and myeloid antigens at the same time as the MPO activity. RESULTS--Six of the 14 cases were initially classified as acute lymphoblastic leukaemia (ALL) and eight as acute myeloid leukaemia (AML). MPO activity was shown at the ultrastructural level in 4-99% blasts from all cases. Six of the 14 were MPO negative by light microscopy and three of these were negative with the antibody anti-MPO. Coexpression of lymphoid antigens (CD19, CD10, or CD2) and MPO was shown by the immunogold method in four out of 11 cases; in seven cases the blasts coexpressed myeloid antigens (CD13, CD33) and MPO. CONCLUSIONS--Electron microscopy is more sensitive for showing MPO than light microscopy and immunocytochemistry; the immunogold method combined with MPO used at the ultrastructural level can help to define the cell lineage involved in biphenotypic leukaemia by highlighting the myeloid component defined by MPO.

Full text

PDF
903

Images in this article

Selected References

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

  1. Akashi K., Shibuya T., Harada M., Morioka E., Oshima K., Kimura N., Takeshita M., Kurokawa M., Kikuchi M., Niho Y. Acute 'bilineal-biphenotypic' leukaemia. Br J Haematol. 1990 Apr;74(4):402–407. doi: 10.1111/j.1365-2141.1990.tb06326.x. [DOI] [PubMed] [Google Scholar]
  2. Bennett J. M., Catovsky D., Daniel M. T., Flandrin G., Galton D. A., Gralnick H. R., Sultan C. Proposals for the classification of the acute leukaemias. French-American-British (FAB) co-operative group. Br J Haematol. 1976 Aug;33(4):451–458. doi: 10.1111/j.1365-2141.1976.tb03563.x. [DOI] [PubMed] [Google Scholar]
  3. Breton-Gorius J., Vanhaeke D., Pryzwansky K. B., Guichard J., Tabilio A., Vainchenker W., Carmel R. Simultaneous detection of membrane markers with monoclonal antibodies and peroxidatic activities in leukaemia: ultrastructural analysis using a new method of fixation preserving the platelet peroxidase. Br J Haematol. 1984 Nov;58(3):447–458. doi: 10.1111/j.1365-2141.1984.tb03991.x. [DOI] [PubMed] [Google Scholar]
  4. Buccheri V., Matutes E., Dyer M. J., Catovsky D. Lineage commitment in biphenotypic acute leukemia. Leukemia. 1993 Jun;7(6):919–927. [PubMed] [Google Scholar]
  5. Campana D., Hansen-Hagge T. E., Matutes E., Coustan-Smith E., Yokota S., Shetty V., Bartram C. R., Janossy G. Phenotypic, genotypic, cytochemical, and ultrastructural characterization of acute undifferentiated leukemia. Leukemia. 1990 Sep;4(9):620–624. [PubMed] [Google Scholar]
  6. Cross A. H., Goorha R. M., Nuss R., Behm F. G., Murphy S. B., Kalwinsky D. K., Raimondi S., Kitchingman G. R., Mirro J., Jr Acute myeloid leukemia with T-lymphoid features: a distinct biologic and clinical entity. Blood. 1988 Aug;72(2):579–587. [PubMed] [Google Scholar]
  7. 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]
  8. Guyotat D., Campos L., Shi Z. H., Charrin C., Treille D., Magaud J. P., Fiere D. Myeloid surface antigen expression in adult acute lymphoblastic leukemia. Leukemia. 1990 Sep;4(9):664–666. [PubMed] [Google Scholar]
  9. Heil G., Gunsilius E., Raghavachar A., Bartram C. R., Ganser A., Kurrle E., Aydemir U., Löffler H., Hoelzer D., Thiel E. Ultrastructural demonstration of peroxidase expression in acute unclassified leukemias: correlation to immunophenotype and treatment outcome. Blood. 1991 Mar 15;77(6):1305–1312. [PubMed] [Google Scholar]
  10. Kantarjian H. M., Hirsch-Ginsberg C., Yee G., Huh Y., Freireich E. J., Stass S. Mixed-lineage leukemia revisited: acute lymphocytic leukemia with myeloperoxidase-positive blasts by electron microscopy. Blood. 1990 Aug 15;76(4):808–813. [PubMed] [Google Scholar]
  11. Lavabre-Bertrand T., Vannereau H., Donadio D., Escola M. J., Taib J., Poncelet P., Rousset T., Bertheault F., Lefort G., Senelar R. Interest of simultaneous ultrastructural characterization of morphology, cytochemistry and immune phenotype in a case of putative hybrid acute leukaemia. Eur J Haematol. 1989 Feb;42(2):202–206. doi: 10.1111/j.1600-0609.1989.tb01211.x. [DOI] [PubMed] [Google Scholar]
  12. Marie J. P., Vernant J. P., Dreyfus B., Breton-Gorius J. Ultrastructural localization of peroxidases in 'undifferentiated' blasts during the blast crisis of chronic granulocytic leukaemia. Br J Haematol. 1979 Dec;43(4):549–558. doi: 10.1111/j.1365-2141.1979.tb03787.x. [DOI] [PubMed] [Google Scholar]
  13. Matutes E., Pombo de Oliveira M., Foroni L., Morilla R., Catovsky D. The role of ultrastructural cytochemistry and monoclonal antibodies in clarifying the nature of undifferentiated cells in acute leukaemia. Br J Haematol. 1988 Jun;69(2):205–211. doi: 10.1111/j.1365-2141.1988.tb07623.x. [DOI] [PubMed] [Google Scholar]
  14. Murphy S. B., Stass S., Kalwinsky D., Rivera G. Phenotypic conversion of acute leukaemia from T-lymphoblastic to myeloblastic induced by therapy with 2'-deoxycoformycin. Br J Haematol. 1983 Oct;55(2):285–293. doi: 10.1111/j.1365-2141.1983.tb01249.x. [DOI] [PubMed] [Google Scholar]
  15. Neame P. B., Soamboonsrup P., Browman G., Barr R. D., Saeed N., Chan B., Pai M., Benger A., Wilson W. E., Walker I. R. Simultaneous or sequential expression of lymphoid and myeloid phenotypes in acute leukemia. Blood. 1985 Jan;65(1):142–148. [PubMed] [Google Scholar]
  16. Polli N., Lambertenghi-Deliliers G., Schirò R., Cattoretti G., Soligo D., Cantù-Rajnoldi A., Romitti L., Delia D., Polli E. E. Relevance of ultrastructural immunocytochemistry in the characterization of unclassifiable leukemias: correlation with phenotypic and genic studies. Haematologica. 1989 Mar-Apr;74(2):129–136. [PubMed] [Google Scholar]
  17. Polli N., O'Brien M., Tavares de Castro J., Matutes E., San Miguel J. F., Catovsky D. Characterization of blast cells in chronic granulocytic leukaemia in transformation, acute myelofibrosis and undifferentiated leukaemia. I. Ultrastructural morphology and cytochemistry. Br J Haematol. 1985 Feb;59(2):277–296. doi: 10.1111/j.1365-2141.1985.tb02994.x. [DOI] [PubMed] [Google Scholar]
  18. Pui C. H., Behm F. G., Kalwinsky D. K., Murphy S. B., Butler D. L., Dahl G. V., Mirro J. Clinical significance of low levels of myeloperoxidase positivity in childhood acute nonlymphoblastic leukemia. Blood. 1987 Jul;70(1):51–54. [PubMed] [Google Scholar]
  19. Pui C. H., Dahl G. V., Melvin S., Williams D. L., Peiper S., Mirro J., Murphy S. B., Stass S. Acute leukaemia with mixed lymphoid and myeloid phenotype. Br J Haematol. 1984 Jan;56(1):121–130. doi: 10.1111/j.1365-2141.1984.tb01277.x. [DOI] [PubMed] [Google Scholar]
  20. Roels F., Wisse E., De Prest B., van der Meulen J. Cytochemical discrimination between catalases and peroxidases using diaminobenzidine. Histochemistry. 1975;41(4):281–312. doi: 10.1007/BF00490073. [DOI] [PubMed] [Google Scholar]
  21. Stass S., Mirro J., Melvin S., Pui C. H., Murphy S. B., Williams D. Lineage switch in acute leukemia. Blood. 1984 Sep;64(3):701–706. [PubMed] [Google Scholar]
  22. Vainchenker W., Villeval J. L., Tabilio A., Matamis H., Karianakis G., Guichard J., Henri A., Vernant J. P., Rochant H., Breton-Gorius J. Immunophenotype of leukemic blasts with small peroxidase-positive granules detected by electron microscopy. Leukemia. 1988 May;2(5):274–281. [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES