Which compartments are involved in Philadelphia‐chromosome positive chronic myeloid leukaemia? An answer at the single cell level by combining May‐Grünwald‐Giemsa staining and fluorescence in situ hybridization techniques

Abstract

Summary. Chronic myeloid leukaemia (CML) is believed to represent a stem cell disorder involving all three cell lineages. The typical chromosomal aberration, the Philadelphia chromosome, is the translocation (9;22)(q34;q11). Several studies with cytogenetics, fluorescence in situ hybridization (FISH), or polymerase chain reaction have investigated the presence of the t(9;22) in different cell compartments. However, questions still remain. In six cases of CML we combined the standard May-Grü nwald-Giemsa staining with FISH at the single-cell level and were able to demonstrate that not only all maturation stages of granulopoiesis, erythropoiesis, and megakaryocytes, but also plasma cells, eosinophils, basophils and monocytes carried the Philadelphia chromosome in 53-98% of samples. Using immunological identification of single cells we were able to demonstrate that the t(9;22) is detectable in 34% of CD3-positive T lymphocytes, in 32% of CD19-positive B lymphocytes, and in 82% of CD34-positive precursor cells. The results give new insight into the biology of CML and may have implications for future therapeutic strategies.

Aberrant cytokine signaling in leukemia

“…In CML, the LSC appear to be very similar if not identical to normal HSC, with multilineage differentiation capacity in patients (Haferlach et al, 1997) and multilineage repopulating activity in NOD-SCID mice Eisterer et al, 2005). In mouse models, whereas transgenic expression of BCR-ABL in CMP causes a CML-like phenotype (Jaiswal et al, 2003), retroviral expression of BCR-ABL in purified CMP or GMP does not produce MPD in recipient mice (Huntly et al, 2004).…”

Fluorescence in situ hybridization for the detection and monitoring of the Ph-positive clone in chronic myelogenous leukemia: comparison with metaphase banding analysis

“…Similar findings were recently reported by Muhlman et al, 23 who calculated the proportion of BCR/ABL + nuclei relative to the percentage of myeloid elements, in order to reduce counting errors deriving from the presence of B or T lymphocytes. The involvement of lymphocytes in CML is a matter of controversy and this issue was recently studied and reviewed by Haferlach and colleagues, 24 who showed that 25-50% of lymphocytes carried BCR/ABL fusion in CML chronic phase.…”

Simple and reliably sensitive diagnosis and monitoring of Philadelphia chromosome-positive cells in chronic myeloid leukemia by interphase fluorescence in situ hybridization of peripheral blood cells

“…2,[42][43][44][45] There is some controversy over whether lymphocytes are of leukemia origin in CML. Garicochea et al 46 reported that T lymphocytes had no bcr/abl fusion gene, while Tefferi et al 42 demonstrated that t(9;22) was detectable in 34% of CD3-positive T lymphocytes, in 32% of CD19-positive B lymphocytes and in 82% of CD34-positive precursor cells.…”