In AML t(8;21) colony growth of both leukemic and residual normal progenitors is restricted to the CD34+ lineage-negative fraction

In an earlier study we observed residual normal colonies in the CD34(+), lineage-negative fraction in AML with a differentiated phenotype. The phenotype of both normal and leukemic progenitors in AML M2, t(8;21) was the subject of this study. The specific translocation enabled discrimination of normal and leukemic cells. Bane marrow samples from eight patients were evaluated for CD34 and the differentiation markers CD33, CD19 and CD56. Growth in all phenotypic fractions was measured in a single cell assay, which enabled quantification of plating efficiency, colony size and determination of progenitor cell origin. No growth was observed In the CD34-negative fraction. In the CD34(+), lineage-positive fraction only clusters up to 20 cells were found in 6/8 samples. In 7/8 samples highly proliferative myeloid, erythroid and mixed colonies were cloned from the CD34(+)/CD56(-)CD19(-)CD33(-) fraction with a frequency between 1 and 12%. Such large colonies grew at a lower frequency (1-6%) from the CD34(+)/CD56(-) fraction (4/8 samples), the CD34(+)/CD56(-)CD19(-) fraction (5/8 samples) and from the CD34(+)CD19(-) fraction (1/8 samples), respectively. Among the colonies consisting of more than 150 cells, only 3/45 evaluated were positive for the AML1/ETO fusion transcript. On the other hand, 8/19 colonies with less than 150 cells were AML1/ETO positive. This study shows that like normal progenitors leukemic progenitors are also present exclusively in the lineage-negative fraction in AML M2 1(8;21). A similar hierarchy of proliferation and differentiation was found for these leukemic progenitors, the smaller colony size fitting with their limited proliferation capacity. The frequency of leukemic progenitors was in the same range as their normal counterparts and detectable only after enrichment for the CD34(+), lineage-negative population.