Flow cytometric analysis of BCL-2 can distinguish small numbers of acute lymphoblastic leukaemia cells from B-cell precursors

Abstract

Flow cytometric identification of small numbers of precursor B-cell acute lymphoblastic leukaemia (B-ALL) cells in post-treatment marrow specimens could benefit from the identification of additional, easily detectable markers that could be used in most cases. In this study, we evaluate whether bcl-2 expression quantified by four-colour flow cytometry can be effectively used to discriminate precursor B-ALL blasts from normal B-cell precursors (haematogones) and function as a leukaemia-specific marker. Levels of bcl-2 in the 22 precursor B-ALL cases studied were found to be significantly higher (over sixfold higher on average) than those present in haematogone populations from 22 control marrow specimens. Higher relative levels of bcl-2 expression in the B-ALL cases were maintained with respect to both immature CD34+ and more mature CD34- haematogone subsets, and appeared stable. Dilutional studies indicated that multiparameter flow cytometry analysis using bcl-2 could identify precursor B-ALL blasts representing as few as 1% of CD19+ cells or 0.01% of total leucocytes in bone marrow specimens containing substantial numbers of haematogones. This study suggests that bcl-2 may be an important marker for flow cytometric detection and quantification of small numbers of residual precursor B-ALL cells in bone marrow specimens.