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Blood, 15 June 2001, Vol. 97, No. 12, pp. 3925-3930
NEOPLASIA
Detection of leukemic cells in the
CD34+CD38 bone marrow progenitor population
in children with acute lymphoblastic leukemia
Aswathi A. George,
Janet Franklin,
Keith Kerkof,
Ami J. Shah,
Mary Price,
Eleanor Tsark,
David Bockstoce,
Dapeng Yao,
Nancy Hart,
Sherri Carcich,
Robertson Parkman,
Gay M. Crooks, and
Kenneth Weinberg
From the Divisions of Research Immunology/Bone Marrow
Transplantation and Hematology-Oncology, Childrens Hospital, Los
Angeles, CA.
Successful autologous hematopoietic stem cell (HSC) transplantation
in childhood acute lymphoblastic leukemia (ALL) requires the ability to
either selectively kill the leukemia cells or separate normal from
leukemic HSC. Based on previous studies showing that more than 95% of
childhood B-lineage ALL express CD38, this study evaluated whether
normal CD34+CD38 progenitors from children
with B-lineage ALL could be isolated by flow cytometry.
CD34+ cells from bone marrow samples from 10 children with
B-lineage ALL were isolated at day 28 of treatment, when clinical
remission had been attained. The CD34+ progenitor cells
were flow cytometrically sorted into CD34+CD38+
and CD34+CD38 populations. The absolute
numbers of CD34+CD38 cells that could
be isolated ranged from 401 to 6245. The cells were then analyzed for
the presence of clonotypic rearrangements of the T-cell receptor (TCR)
V 2-D3 locus. Only patients whose diagnostic marrow had an
informative TCR V2-D3 rearrangement were included in this
study. Detection thresholds were typically 104 to
105 leukemic cells in normal marrow. In 6 of 10 samples analyzed, the sorted CD34+CD38 cells
had no detectable V2-D3 rearrangements. In 4 cases, the clonotypic leukemic V2-D3 rearrangement was detected in the CD34+CD38 population, indicating that
the putative normal HSC population also contained leukemic cells. The
data indicate that although most childhood ALL cells express CD34 and
CD38, leukemic cells are also frequently present in the
CD34+CD38 population. Therefore, strategies
to isolate and transplant normal HSC from children with ALL
will require a more stringent definition of the normal HSC than
the CD34+CD38 phenotype.
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