Comparative flow cytometric evaluation of bcl-2 oncoprotein in CD5+ and CD5- B-cell lymphoid chronic leukemias

Haematologica. 1997 Sep-Oct;82(5):555-9.

Abstract

Background and objectives: Levels of intracellular bcl-2 oncoprotein have been found to be increased in leukemic cells of CD5+ B-chronic lymphocytic leukemia (CLL) patients. However, it is not clear whether bcl-2 overexpression is a peculiar feature of CD5+ B-CLL. Based on this background we carried out a quantitative flow cytometric evaluation of intracellular bcl-2 levels on leukemic cells of CD5+ and CD5- B-CLL.

Methods: We assessed in flow cytometry levels of bcl-2 protein using a quantitative indirect immunofluorescence assay (QIFI kit) on samples from 46 previously untreated CD5+ B-CLL patients. Results were compared with those obtained on either normal peripheral blood B-lymphocytes or leukemic cells from 7 CD5- B-CLL patients intentionally selected for statistical comparison.

Results: A relatively homogeneous amount of bcl-2 protein which did not reflect either clinical-biological features at the time of diagnosis nor in vivo response to therapy was found. Results expressed as antibody binding capacity (ABC) accounted for a mean value of 12.2 +/- 1.5 x 10(3) molecules/cell (range, 6.4-13 x 10(3) molecules/cell). Levels of bcl-2 detected on CD5+ B-CLL leukemic cells were significantly lower than those of B peripheral blood lymphocytes from healthy donors (p = 0.0001). The same applied when comparing CD5+ and CD5- B-CLL patients (bcl-2 ABC, 8.07 +/- 0.26 x 10(3) molecules/cell vs. 12.2 +/- 1.5 x 10(9) molecules/cell; p = 0.0001).

Interpretation and conclusions: According to the role of bcl-2 in preventing apoptosis, our results indicate that differences in the pattern of expression of such an oncoprotein, might, at least in part, explain the more aggressive clinical course of CD5- B-CLL forms.

Publication types

  • Comparative Study

MeSH terms

  • Aged
  • B-Lymphocytes / chemistry*
  • CD5 Antigens / analysis*
  • Cell Separation
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism*
  • Male
  • Middle Aged
  • Proto-Oncogene Proteins c-bcl-2 / analysis*
  • Reagent Kits, Diagnostic

Substances

  • CD5 Antigens
  • Proto-Oncogene Proteins c-bcl-2
  • Reagent Kits, Diagnostic