The Effect of Bcr-Abl Protein Tyrosine Kinase on Maturation and Proliferation of Primitive Haematopoietic Cells

Abstract

Background: Chronic Myeloid Leukaemia (CML) is characterised by the chromosomal translocation resulting in expression of the Bcr-Abl protein tyrosine kinase (PTK) in early stem cells and their progeny. However the precise nature of Bcr-Abl effects in primitive CML stem cells remains a matter of active debate. Materials and Methods: Extremely primitive Bcr-Abl fusion positive cells were purified from patients with CML using multiparameter flow cytometric analysis of CD34, Thy, and lineage marker (Lin) expression, plus rhodamine-123 (Rh-123) brightness. Progenitor cells of increasing maturity were examined for cycling status by flow cytometry and their proliferative status directly correlated with cell phenotype. The activation status of a key transcription factor, signal transducers and activators of transcription (STAT-5), was also analyzed by immunocytochemistry. Results: The most primitive stem cells currently defined (CD34+Lin-Thy+ Rh-123lo) were present as a lower proportion of the stem cell compartment (CD34+Lin-) of CML patients at presentation than of normal individuals (2.3% ± 0.4 compared with 5.1% ± 0.6 respectively). Conversely there was a significantly higher proportion of the more mature cells (CD34+Lin-Thy-Rh-123 hi) in CML patients than in normal individuals (79.3 ± 1.8 compared with 70.9 ± 3.3). No primitive subpopulation of CML CD34+Lin-cells was cycling to a significantly greater degree than cells from normal donors, in fact, late progenitor cells (CD34+ Lin+) were cycling significantly less in CML samples than normal samples. STAT5, however, was observed to be activated in CML cells. Conclusions: We conclude that no subpopulation of CML stem cells displays significantly increased cell cycling. Thus, increased cycling cannot be a direct consequence of Bcr-Abl PTK acquisition in highly enriched stem cells from patients with CML. In vivo CML need not be considered a disease of unbridled stem cell proliferation, but a subtle defect in the balance between self renewal and maturation.