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Blood, 15 June 2001, Vol. 97, No. 12, pp. 3882-3889
NEOPLASIA
Hoechst 33342 efflux identifies a subpopulation of
cytogenetically normal CD34+CD38 progenitor
cells from patients with acute myeloid leukemia
Michaela Feuring-Buske and
Donna E. Hogge
From the Terry Fox Laboratory, British Columbia Cancer
Agency, Vancouver, Canada.
Efflux of Hoechst 33342 from normal hematopoietic cells identifies
a "side population" (SP+) of negatively staining cells
that, in the mouse, are largely CD34 and are enriched for
primitive progenitors. To further characterize human SP+
cells, blood or bone marrow from 16 patients with acute myeloid leukemia (AML) was analyzed for their presence, immunophenotype, and
cytogenetic and functional properties, and for the relation between SP
phenotype and multidrug resistance-1 (MDR-1) expression. The
mean percentages of SP+ and MDR+ cells was
8.1% (range, 0.5%-29.9%) and 12.8% (range, 0%-54.8%), respectively, with no correlation between the 2 values. The percentages of SP+ cells that were CD34+CD38,
CD34+CD38+, or CD34 were 12%
(range, 0.4%-50%), 25% (range, 0.5%-96%), and 63% (range, 4%-99%). Cytogenetically abnormal cells were always detected in the
SPCD34+CD38 and
SP+CD34 fractions, and abnormal colonies
(CFC), long-term culture-initiating cells (LTC-IC), and nonobese
diabetic-severe combined immunodeficiency (NOD/SCID) mouse leukemia-IC
were detected in the former fraction. No progenitors were detected
among SP+CD34 cells in any of these assays
from 9 of 10 samples. In contrast, exclusively normal cells were
detected in the SP+CD34+CD38
fraction from 9 of 15 samples, and CFC, LTC-IC, and multilineage engraftment in NOD/SCID mice from this subpopulation were also cytogenetically normal in 6 of 8, 6 of 7, and 2 of 2 cases studied, respectively. In contrast to murine studies, primitive progenitors are
enriched among SP+CD34+CD38 cells
from patients with AML. The molecular basis for Hoechst dye efflux is
uncertain because it does not appear to be related to MDR-1 expression.
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