Different Potential of G-CSF, GM-CSF, and pIXY in Priming of Myeloid Leukemic Progenitors for Ara-C
Cell kinect or metabolic resistance of myeloid leukemic cells to cycle-specific cytostatics, such as cytosine arabinoside (ara-C), may be overcome by priming with growth factors. The extent of priming with GM-CSF and IL-3 has been highly variable for meyloid leukemic progenitors in previous studies. We tested nine different AML samples for their ara-C sensitivity and the priming effects of G-CSF, GM-CSF, GMCSF plus IL-3 and pIXY321. Bone marrow mononuclear cells were isolated from patients with newly diagnosed AML. All factors were used at 100 U/ml for 48 h, controls were incubated in medium only (IMDM, 10% FBS). Ara-C was added for 12 h at concentrations from 0 to 100 µM. Cells were washed and seeded into a colony assay based on methylcellulose with serum-free conditions using a mixture of hemopoietic growth factors. After 14 days colonies were scored. From the dose-response curves for ara-C, D50 values were calculated with confidence limits and compared for different pretreatments, showing shifts in ara-C sensitivity due to priming. D50 values for ara-C covered a range of 1 log, in previous studies over 2 logs with a larger sample size. Priming with various factors decreased the D50 values for the majority of samples, The largest shifts were obtained with pIXY321 or the equimolar mixture of GM-CSF and IL-3 at a factor of 1:1500. Samples unresponsive to priming with one growht factor responded to another factor. Therefore, priming with a combination of factors, triggering different signal transduction pathways, might be more promising.
KeywordsAcute Myeloid Leukemia Chronic Myeloid Leukemia Cytosine Arabinoside Bone Marrow Mononuclear Cell Colony Assay
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