CD34 Quantitative Flow Cytometry Study of the Blastic Population in Acute Leukemias

  • Gian Matteo Rigolin
  • Francesco Lanza
  • Gianluigi Castoldi


Quantitative flow cytometry analysis of membrane antigens is becoming a routinely approach in the analysis of the blastic population in acute leukemias. By flow cytometry (FACScan B.D.), we analysed CD34 quantitative expression on the blastic population of 48 patients affected by acute leukemias (39 AML, 9 ALL). The blastic population was defined by means of strategical double fluorescence combinations of monoclonal antibodies. Patients were subdivided into four groups: de novo AML (24 pts), secondary AML (15 pts), relapses of AML (8 pts) and ALL (9 pts). A significant higher intensity of expression was demonstrated in secondary AML (p=0.019) and at relapses (p=0.026). At relapse, a higher percentage of CD34 positivity and a higher intensity of CD34 expression were observed. When considering AML FAB distribution a significant higher CD34 intensity of expression was demonstrated in M0, M1 and M4 (p=0.003). When subdividing patients according to negative, weak and bright CD34 intensity of expression, the 93.3% of secondary AML had a bright, expression while only a 50% of de novo AML was recorded in the bright CD34 group. Only one secondary AML had negative CD34 expression while the 33.3% of de novo AML was CD34 negative (p=0.02). Secondary AML demonstrated a lower CD33 and a higher HLA-DR intensity of expression. According this quantitative CD34 analysis and considering the models of immunological development of hemopoiesis, we may prospect that in secondary AML the neoplastic event prevalently involve a stem cell at an earlier stage of development. We think that from the combination of more antigens on a larger series of patients and considering a combined immunological and cytogenetic study we may confirm the important role of flow cytometry in the study of the heterogeneity of immunological findings in acute leukemias with biological and prognostic implications.


Acute Myeloid Leukemia Acute Lymphoblastic Leukemia Acute Leukemia Acute Myeloid Leukemia Patient CD34 Percentage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Terstappen LWMM, Safford M, Könemann S, Loken MR, Zurlutter K, Büchner T, Hiddemann W, Wörmann B: Flow cytometric characterisation of acute myeloid leukemia. Part II. Phenotypic heterogeneity at diagnosis. Leukemia 5: 757, 1991.Google Scholar
  2. 2.
    Farahat N, Lens D, Matutes E, Catovsky D: Differential TdT expression in acute leukemia by flow cytometry: a quantitative study. Leukemia 9: 583, 1995.PubMedGoogle Scholar
  3. 3.
    Lavabre-Bertrand T, Janossy G, Ivory K, Peters R, Seeker-Walker L, Poerwit-MacDonald A: Leukemia-associated changes identified by quntitative flow cytometry: CD 10 expression. Cytometry 18: 209, 1994.PubMedCrossRefGoogle Scholar
  4. 4.
    Bennett JM, Catowsky D, Daniel MT, Flandrin G, Galton DAG, Gralnick HR, Sultan C: Proposed revised criteria for the classification of acute myeloid leukemia. Ann Intern Med 103: 626, 1985.Google Scholar
  5. 5.
    Castoldi GL, Cuneo A, Lanza F, Tomasi P: Diagnosis of leukemia: morphology. Leukemia 6;Suppl. 4: 6, 1992.Google Scholar
  6. 6.
    Castoldi GL, Liso V, Fenu S, Vegna L, Mandelli F: Reproducibility of the morphological diagnostic criteria for acute myeloid leukemia: the GIMEMA group experience. Ann Hematol 66: 171, 1993.PubMedCrossRefGoogle Scholar
  7. 7.
    Terstappen LWMM, Könemann S, Safford M, Loken MR, Zurlutter K, Büchner T, Hiddemann W, Wörmann B: Flow cytometric characterization of acute myeloid leukemia. Part I. significance of light scattering properties. Leukemia 5: 315, 1991.PubMedGoogle Scholar
  8. 8.
    Verwer BJH, Terstappen LWMM: Automatic lineage assignment of acute leukemias by flow cytometry. Cytometry 14: 862, 1993.PubMedCrossRefGoogle Scholar
  9. 9.
    Kawada H, Ichikawa Y, Watanabe S, Nagao T, Arimori S: Flow cytometry analysis of cell-surface antigen expressions on acute myeloid leukemia cell populations according to their cell-size. Leukemia Res 18: 29, 1994.CrossRefGoogle Scholar
  10. 10.
    Kawada H, Ichikawa Y, Watanabe S, Nagao T, Arimori S: Flow cytometry analysis of cell-surface antigen expressions on acute myeloid leukemia cell populations according to their cell-size. Leukemia Res 18: 29, 1994.CrossRefGoogle Scholar
  11. 11.
    Lanza F, Rigolin GM, Moretti S, Latorraca A, Castoldi GL: Prognostic value of immunophenotypic characteristics of blast cells in acute myeloid leukemia. Leukemia Lymphoma 1994; 13 suppl 1:81–5.PubMedCrossRefGoogle Scholar
  12. 12.
    Terstappen LWMM, Safford M, Unterhalt M, Könemann S, Zurlutter K, Piechotka K, Drescher M, Aul C, Büchner T, Hiddemann W, Wörmann B: Flow cytometric characterization of acute myeloid leukemia: IV. Comparison to the differentiation pathway of normal haematopoietic progenitors cells. Leukemia 6: 993, 1992.PubMedGoogle Scholar
  13. 13.
    Pirelli L, Teofili L, Menichella G, Rumi C, Paoloni A, Iovino S, Puggioni PL, Leoni G, Bizzi,B: Further investigations on the expression of HLA-DR, CD33 and CD 13 surface antigens in purified bone marrow and peripheral blood CD34+ haematopoietic progenitor cells. Br J Haematol 84: 24, 1993.CrossRefGoogle Scholar
  14. 14.
    Pontvert-Delucq S, Breton-Gorius J, Schmitt C, Baillou C, Guichard J, Najman A, Lemoine M: Characterization and functional analysis of adult human bone-marrow cell subsets in relation to B-lymphoid development. Blood 82: 417, 1993.PubMedGoogle Scholar
  15. 15.
    Terstappen LWMM, Huang S, Safford M, Lansdorp M, Loken MR: Sequential generation of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitors cells. Blood 77: 1218, 1991.PubMedGoogle Scholar
  16. 16.
    Campos L, Guyotat D, Archimbaud E, Devaux Y, Treille D, Larese A, Maupas J, Gentilhomme O, Ehrsam A, Fiere D: Surface marker expression in adult acute myeloid leukemia: correlations with initial characteristics, morphology and response to therapy. Br J Haematol 72: 161, 1989.PubMedCrossRefGoogle Scholar
  17. 17.
    Geller R, Zahurak M, Hurwitz A, Burke PJ, Karp JE, Piantadosi S, Civin CI: Prognostic importance of immunophenotyping in adults with acute myelocitic leukemia: the significance of the stem-cell glycoprotein CD34 (My 10). Br J Haematol 76: 340, 1990.PubMedCrossRefGoogle Scholar
  18. 18.
    Borowitz MJ, Guenther KL, Shults KE, Stelzer GT: Immunophenotyping of acute leukemia by flow cytometric analysis. Am J Clin Pathol 100, 534, 1993.PubMedGoogle Scholar
  19. 19.
    Solary E, Casanovas O, Campos L, Béné MC, Faure G, Maingon P, Falkenrodt A, Lenhormand B, Genetet N and GEIL: Surface markers in adult myeloblastic leukemia: correlation of CD19+, CD34+ and CD14+/DR- phenotypes with shorter survival. Leukemia 6: 393, 1992.PubMedGoogle Scholar
  20. 20.
    Del Poeta G, Stasi R, Venditti A, Suppo G, Aronica G, Bruno A, Masi M, Tabilio A, Papa G: Prognostic value of cell marker analysis in de novo acute myeloid leukemia. Leukemia 8: 388, 1994.PubMedGoogle Scholar
  21. 21.
    Bradstock K, Mattews J, Benson E, Page F, Bishop J, and the Australian leukemia study group: Prognostic value of immunophenotyping in acute myeloid leukemia >Blood 84: 1220, 1994.Google Scholar
  22. 22.
    Paietta E, Andersen J, Gallagher R, Bennett J, Yunis J, Cassileth P, Rowe J, Wiernik PH: The immuno-phenotype of acute promyelocytic leukemia (APL): an ECOG study. Leukemia 8: 1108, 1994PubMedGoogle Scholar
  23. 23.
    Kita K, Nakase K, Miwa H, Masuya M, Nishii K, Morita N, Takakura N, Otsuji A, Shirakawa S, Ueda T, Nasu K, Kyo T, Dohy H, Kamada N: Phenotypical characteristics of acute myelocytic leukemia associated with the t(8;21)(q22;q22) chromosomal abnormality: frequent expression of immature B-cell antigen CD 19 togheter with stem cell antigen CD34. Blood 80: 470, 1992.PubMedGoogle Scholar
  24. 24.
    Thomas X, Campos L, Archimbaud E, Shi ZH, Ritoutet T, Anglaret B, Fiere D: Surface marker expression in acute myeloid leukemia at first relapse. Br J Hematol 81: 40, 1992.CrossRefGoogle Scholar
  25. 25.
    Fagioli F, Cuneo A, Carli MG, Bardi A, Piva N, Previati R, Rigolin GM, Ferrari L, Spanedda R, Castoldi GL: (1993) Chromosome aberratios in CD34-positive acute myeloid leukemia. Correlation with clinico-pathologic features. Cancer Genet Cytogenet 71: 119, 1993.Google Scholar
  26. 26.
    Rigolin GM, Lanza F, Castoldi GL: Photomultiplier voltage setting: possible important source of variability in MESF calculation? Cytometry (in press).Google Scholar

Copyright information

© Plenum Press, New York 1996

Authors and Affiliations

  • Gian Matteo Rigolin
  • Francesco Lanza
    • 1
  • Gianluigi Castoldi
    • 1
  1. 1.Institute of HematologyUniversity of FerraraFerraraItaly

Personalised recommendations