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Blood, Vol. 92 No. 12 (December 15), 1998:
pp. 4758-4763
Lineage Involvement of Stem Cells Bearing the Philadelphia
Chromosome in Chronic Myeloid Leukemia in the Chronic Phase as
Shown by a Combination of Fluorescence-Activated Cell Sorting and
Fluorescence In Situ Hybridization
By
Naoto Takahashi,
Ikuo Miura,
Kohki Saitoh, and
Akira B. Miura
From the Third Department of Internal Medicine, Akita University
School of Medicine, Akita, Japan.
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ABSTRACT |
Chronic myeloid leukemia (CML) is thought to arise from a
pluripotent hematopoietic stem cell that has undergone a reciprocal translocation between the BCR gene on chromosome 22 and the ABL proto-oncogene on chromosome 9. This rearrangement results in a
shortened chromosome 22, designated the Philadelphia (Ph) chromosome. The Ph chromosome has been found in cells from all hematopoietic lineages except mature T lymphocytes. To examine this issue, we combined fluorescence-activated cell sorting (FACS) and fluorescence in
situ hybridization (FISH) to study lineage involvement of mature cells
and stem cells in 12 patients with CML in the chronic phase. We found
Ph chromosomes in myeloid cells and most B lymphocytes (CD19+) but not in mature T cells (CD3+) or
natural killer (NK) cells (CD3 56+).
Moreover, evidence of BCR/ABL fusion was found in pluripotent stem cells (CD34+Thy-1+), B-progenitor
cells (CD34+CD19+), T/NK progenitor
cells (CD34+CD7+ cells), and T progenitor
cells (CD34+CD7+CD5+) with a
frequency equal to that in all CD34+ cells isolated by
FACS from bone marrow cells. T lymphocytes showed a marked decrease
in Ph+ cells between progenitor cells and mature cells.
Moreover, the ratios of Ph+ to Ph cells in
mature T cells and NK cells were below background levels, whereas
Ph+ B lymphocytes also decreased during their
maturation. These data suggest that Ph+ lymphocytes are
eliminated during differentiation. In contrast to FISH of blood and
bone marrow, which gives information principally about mature cells,
the technique of "sorter FISH (FACS + FISH)" provides a
powerful tool to explore the cytogenetic changes in immature cell
populations of stem cell diseases based on immunophenotypes. Further
clarification of genetic changes in stem cells could be achieved by
using sorter FISH with monoclonal antibodies.
© 1998 by The American Society of Hematology.
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INTRODUCTION |
CHRONIC myelogenous leukemia (CML) is a
malignant hematological disorder of the human hematopoietic stem cells.
Studies with glucose-6-phosphate dehydrogenase (G6PD)
isoenzymes1,2 have shown that these stem cells are capable
of differentiation into myeloid cells, monocytes, erythrocytes,
platelets, and B lymphocytes. The Philadelphia (Ph) chromosome,
t(9;22)(q34;q11), is the cytogenetic hallmark of CML; this balanced
translocation results in a chimeric BCR/ABL gene that expresses an
8.5-kb hybrid mRNA transcript and a 210-kD fusion protein (P210). This
BCR/ABL expression is presumed to effect clonal expansion in CML by
deregulation of cell proliferation.3 As a recent advance in
the treatment of CML, interferon- has been found to reduce the
Ph+ clone and prolong survival of CML patients. However,
most patients experience transformation from the chronic phase to an
acute blastic leukemia (blast crisis) after various time periods. In a
majority of patients, the blasts resemble acute myeloblastic leukemia
cells, and in about one third the blasts morphologically resemble the lymphoblasts in acute lymphoblastic leukemia.4 Less
commonly, the dominant phenotype is an erythroblast, a megakaryoblast,
or an undifferentiated blast with no detectable lineage-associated markers.5 T-cell markers in CML blast crisis have rarely
been reported.6,7 This observation supports the hypothesis
that the oncogenic event in CML occurs in a pluripotent hematopoietic stem cell capable of multilineage differentiation.
Acquisition of the Ph chromosome in nonlymphoid cells has been
previously observed. For lack of direct evidence, the lineage involvement of T/natural killer (NK) and B lymphocytes remains unclear.
However, in most studies, direct analysis of T cells showed no evidence
of the Ph chromosome.8-13 In two studies, occurrence of the
Ph chromosome in phytohemagglutinin (PHA)-responding peripheral blood
(PB) cells, mostly peripheral T cells, was reported.14,15 In one study, direct determination of the Ph chromosome in B and T
lymphocytes by means of tricolor immunophenotyping/fluorescence in situ
hybridization (FISH) showed that two patients featured involvement of
CD20+ lymphocytes and that CD3+ lymphocytes in
all six patients were negative for the Ph chromosome.16,17 In another study, T-cell clones from PB cells in the chronic phase were
screened for BCR/ABL fusion transcripts by using reverse transcriptase-polymerase chain reaction (RT-PCR). The
BCR/ABL transcripts could be detected in only a few T-cell
clones.18 Most B-lymphoblastoid cell lines established from
CML patients do not carry the Ph marker or any submicroscopic BCR/ABL
rearrangement.19 Recently, Haferlach et al17
reported that t(9;22) was detectable in 34% of CD3+ T
lymphocytes, in 32% of CD19+ B lymphocytes, and in 82% of
CD34+ progenitor cells when they used a combination of
blood and bone marrow (BM) smears stained with
May-Grünwald-Giemsa and FISH. The Ph+ ratio of mature
T cells was very high compared with those previously reported.20 As can be seen from these findings, the
involvement of lymphocytes still remains to be settled.
This study examines the Ph chromosome status in mature cells and
progenitor cells of various lineages and in multipotent stem cells from
CML patients. The more immature cells are more difficult to culture in
vitro because they are dormant. With fluorescence-activated cell
sorting (FACS), mature cells and stem cells can be sorted on the basis
of immunophenotype, but the number of stem cells is not sufficient for
Southern blot analysis. Moreover, because cell sorting cannot avoid a
few contaminating cells, PCR or cell culture for amplification of the
number of cells would hamper interpretation of the results. Although
chromosome analysis is restricted to metaphase cells, FISH has the
advantage of allowing for a cell-by-cell analysis of the BCR/ABL fusion
signals in nondividing cells that are potentially transcriptionally
inactive. For these reasons, we applied FISH to blood smear (smear
FISH) for mature cells and FACS (sorter FISH) to collected cells for
the stem cells of 12 patients with CML in the chronic phase who had Ph
chromosome in all cells as determined by standard chromosome analysis.
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MATERIALS AND METHODS |
Patients and samples.
The patient characteristics are shown in
Table 1. At the time of study, 4 patients
had not had any prior therapy, but 6 patients had been receiving
chemotherapy with hydroxyurea and 2 patients with interferon- and
hydroxyurea. Ph chromosomes were observed in all analyzed metaphases by
using a standard chromosome analysis technique. PB specimens and BM
were obtained from 12 patients with Ph+ CML in the chronic
phase and from donors of BM transplantation after informed consent was
obtained. BM mononuclear cells (BMMC) or PB mononuclear cells (PBMC)
were isolated by Ficoll-Hypaque density gradient centrifugation.
Phagocytic cells were removed from BMMC or PBMC with Silica Suspension
(Immuno-Biological Laboratories, Gunma, Japan), which enabled us to
collect more CD34+ cells effectively. These nonphagocytic
cells were used for the following study with FACS.
FACS.
Fluorescein-conjugated CD8 (B9.11-FITC), CD7 (3A1-FITC), CD19
(89B-FITC), CDw90 (Thy-1) (F15.42.1.5-FITC), PECy5-conjugated CD34
(581-PECy5), CD4 (13B8.2-PECy5) (Coulter Immunotech, Margency, France),
and fluorescein isothiocyanate (FITC)-conjugated CD3 (Leu4-FITC), HLA-DR-FITC- and phycoerythrin (PE)-conjugated CD34 (HPCA2-PE), CD56 (Leu19-PE) and CD3 (Leu4-PE) (Becton Dickinson, Sunnyvale, CA) were used. Both FITC-conjugated and PE-conjugated nonspecific Ms IgG were obtained from DAKO (Tokyo, Japan).
Flow cytometry analysis and cell sorting were performed on an EPICS
Elite (Coultronics, Margency, France). Between 1,000 and 30,000 cells
per fraction were sorted and collected in fetal calf serum (FCS). The
purity attained was 97%. These cells were used for cytospin
preparations and stained with May-Grünwald-Giemsa.
In situ hybridization.
The probes used in this study were the BCR/ABL translocation probes
commercially available from Oncor, Inc (Gaithersburg, MD). The probes
flank the fusion site in essentially all cases of CML.21
For prehybridization, slides were immersed in 0.01 N HCl/0.001% pepsin
for 5 minutes, washed two times in phosphate-buffered saline (PBS) for
3 minutes, and treated with 4% paraformaldehyde/PBS for 5 minutes.
After being washed with PBS, the cells were dehydrated through 70%,
80%, and 100% ethanol. The hybridization protocol followed the Oncor
instructions and signals were detected with the aid of
fluorescein-labeled avidin/rhodamine-labeled antidigoxigenin (Oncor).
Signals were visualized under a Nikon microscope (Tokyo, Japan) with a
FITC/Rhodamine dual band filter (Nikon). Evaluation of the preparations
was performed by counting 100 nuclei per slide. The mean percentage of
nuclei with a false-positive signal was calculated for the control PB
from hematological disease-free individuals. A previous study reported
false-positive cells occurring at a frequency of about
2%,21 and we found false-positive cells at a frequency of
0% to 6% (cutoff).
Purification of progenitor populations.
Stem cells were characterized by defining subpopulations of
CD34+ cells. Thy-1 is a marker expressed by fetal and adult
BM stem cells and the Thy-1+ subset has multilineage
differentiation capacity, as shown by its ability to produce T cells, B
cells, and myeloid cells.22,23 The
CD34+CD10+CD19+ population
represents exclusively B-lymphoid committed progenitors.24 Cytoplasmic CD3+CD7+ cells were considered to
represent committed T-cell progenitors based on the fact that CD2,
cyCD3, CD5, and CD7 are coexpressed on all mature T cells but not on
all mature B cells.25 However, these antigens are also
expressed on CD34+ NK progenitors.26 In
addition, CD7 is present on B-cell and myeloid
progenitors.27 These data suggest that
CD34+CD7+ and CD5+/CD2+
populations represent T-lymphoid committed progenitors.25
On the basis of these findings, we used
CD34+Thy-1+ cells as the pluripotent stem
cells, CD34+CD19+ as the B progenitor cells,
CD34+CD7+ cells as the T/NK progenitor cells,
and CD34+CD7+CD5+ as the T
progenitor cells in this study.
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RESULTS |
The influence of treatment with interferon- and hydroxyurea on the
proliferation of cell lineage was taken into consideration, but the
data for patients with and without previous treatment at the time of
this study were not different.
Flow cytometry analysis of CD34+ cells obtained from BM
mononuclear cells.
We isolated CD34+ cells from 8 of 12 patients. The other 4 patients were assessed only by FISH of the PB smear.
Table 2 indicates that the number of
CD34+ and CD34+7+ cells was quite
variable and higher than that in normal donors. The other
subpopulations of lymphoid progenitor cells did not differ from those
in normal donors.
Lineage involvement determined by FISH analysis of blood smear.
Neutrophils and monocytes were consistently highly positive for the Ph
chromosome, ie, greater than 90% and 80%, respectively. Because of
the marked increase in granulocytes, we did not find monocytes within
the field observed by FISH in 3 patients (patients no. 6, 8, and 11;
Fig 1).
FISH analysis applied to sorted cells of B, T, and NK cells.
Lymphoid cells were assessed by sorter FISH, which allowed us to
classify lymphocytes on the basis of May-Grünwald-Giemsa staining
into B, T, or NK cells. The percentage of CD19+ B
lymphocytes varied among patients, but all patients had a lower incidence of Ph+ cells among B lymphocytes than among
neutrophils and monocytes (Figs 1 and 2).
In contrast, the incidence of Ph+ cells among
CD3+ T lymphocytes and
CD356+ NK cells was below the cutoff
value in all patients (Fig 3).
FISH analysis of stem cell CD34+ subpopulations.
The incidence of cells bearing the Ph chromosome among
CD34+ cells was as frequent as that seen in other
progenitor cells (Fig 4). Not all
subpopulations could always be isolated with FACS. In contrast to
mature T cells, the incidence of the Ph chromosome in progenitor cells
was similar to that in CD34+ cells. Compared with mature B
cells, the incidence of Ph+ progenitor cells were as
frequent as that in CD34 progenitor cells. The incidence of the Ph
chromosome in CD34+Thy-1+ cells was already
similar to the one seen in mature myeloid cells (Fig 5).
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| Fig 5.
A representative image of FISH applied to
CD34+Thy-1+ and
CD34+7+ cells sorted from BM. (A and C)
May-Giemsa stain of CD34+Thy-1+ and
CD34+7+ cells. (B and D) The hybridized bcr
probe was detected with rhodamine (red signal) and the hybridized abl
probe with fluorescein (yellow-green signal). A yellow or red/green
spot was indicative of BCR/ABL fusion (white arrows).
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Taken all together (Fig 6), the results
from the FISH analysis to detect BCR/ABL fusion show that the Ph
chromosome is present in B lymphocytes (CD19+) but not in
mature T cells (CD3+) or NK cells
(CD3CD56+). Moreover, BCR/ABL fusion was
found in pluripotent stem cells (CD34+Thy-1+;
Fig 6), B progenitor cells
(CD34+CD19+), and T progenitor cells
(CD34+CD7+ or
CD34+CD7+CD5+ cells) at the same
frequency as in all CD34+ cells sorted by FACS from BM
cells. Although neutrophils and monocytes had a high percentage of
Ph+ cells, the ratios of Ph+ to
Ph cells in mature T cells and NK cells were below
background levels. Ph+ B lymphocytes also decreased during
the maturation process.
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| Fig 6.
Ratio of Ph+ cells to Ph
cells. Ph chromosomes were found in myeloid cells and B lymphocytes but
not in mature T and NK cells. BCR/ABL fusion signals were found in
pluripotent stem cells (CD34+Thy-1+), B
progenitor cells (CD34+CD19+), and T
progenitor cells (CD34+CD7+ cells or
CD34+CD7+CD5+) at a frequency
equal to that in all CD34+ cells. T lymphocytes showed a
marked decrease of Ph+ cells between progenitor cells and
mature cells. The incidence ratios of Ph+ cells in mature
T cells and NK cells were below the cutoff value.
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DISCUSSION |
CML is thought to arise from a pluripotent hematopoietic stem cell that
contains the Ph chromosome. The Ph chromosome, t(9;22)(q34;q11), is the
cytogenetic hallmark of CML, and this balanced translocation results in
a chimeric BCR/ABL gene. This BCR/ABL expression is presumed to effect
clonal expansion in CML by means of deregulation of cell proliferation.
FISH applied to blood and BM smear (smear FISH) provided various data
on cell lineage involvement of the Ph chromosome, and these data
provided additional support for the hypothesis that CML is a stem cell
disease. Although smear FISH yielded important information, it could
not provide information on changes affecting pluripotent stem cells or
on progenitor cells and their mature descendants. Smear FISH also has
limitations in assessing polyploid cells and eosinophils because of
their autofluorescence. Cell sorting is a powerful tool to obtain stem
cells of various lineages and progenitor cells of a defined maturation
level. This technique is especially effective for lymphoid cells and
stem cells, because it is difficult to identify subclasses of
lymphocytes based on morphological observations and because immature
cells are dormant and difficult to culture. However, colony assays may
introduce clonal changes during culture periods. Because BCR/ABL mRNA
is minimally expressed or may be absent in primitive CML progenitors, these cells may escape detection by RT-PCR.28 At the same
time, cell fractions obtained by sorting based on immunophenotype may include some contaminated cells that would be amplified by culture or
PCR. As a result of these considerations, we applied FISH to immunophenotype sorted cells for this study.
The Ph chromosome has been found in cells from all hematopoietic
lineages except mature T lymphocytes. Our data summarized in Fig 6 show
that, although mature T lymphocytes do not have the Ph chromosome, both
stem cells and T progenitor cells do. There is thus an obvious
discrepancy in the positivity of the Ph chromosome in T and B
lymphocytes. To resolve this discrepancy between stem cells and mature
lymphocytes, we examined each of the progenitor cells.
To date, several explanations have been proposed for the lack of the Ph
chromosome among peripheral T lymphocytes. A long-standing hypothesis
is that the majority of T cells are long-lived and born before the
occurrence of clonal mutation. It is also believed that memory
lymphocytes are capable of surviving for long periods of 20 years or
more. But two studies in humans have reported the mean life span of
lymphocytes as 530 days29 or 1,600 days,30 indicating that some lymphocytes must have a half-life in excess of 3 years. We must also consider that it takes on average 6.3 years from
the time that a cell acquires the Ph chromosome until a patient has
clinically evident symptoms.31 The present study includes
patients who have had CML for 5 to 10 years (Table 1; patients no. 2, 3, 6, 7, and 9), which is enough time for some T lymphocytes to acquire
the Ph chromosome. Although we found BCR/ABL rearrangement
in progenitor cells committed to the T-cell lineage, no evidence of the
fusion gene was found among peripheral T lymphocytes, even in patients
with a long history of CML. These data lead us to believe that the
lymphocyte life span may not conclusively explain the lack of the Ph
chromosome in mature T lymphocytes.
A second explanation is that the usual target of malignant
transformation in CML is a more restricted stem cell committed to the
myeloid and B-cell lineages but not to the T-cell
lineage.32 In our study, BCR/ABL fusion was detected
in CD34+Thy-1+ cells,
CD34+CD19+ cells, and
CD34+CD7+ or
CD34+CD7+CD5+ cells at the same
frequency as in all CD34 cells. These data indicate that the usual
target of malignant transformation in CML is a pluripotent stem
cell. Thus, a restricted stem cell target may not explain the
negativity of Ph chromosome in T lymphocytes either.
Although the ontogeny of NK cells has not been fully clarified yet,
CD34+ progenitor cells have been shown to differentiate
into NK cells in vitro.26,33-35 Our data show that most NK
cells during the chronic phase of CML are also from
Ph clones.
We must also consider the impact of age on T-cell generation and
turnover. Age-related involution is characterized by a progressive reduction in thymic size and weight. After the first 20 to 30 years of
life, a greater proportion of the thymus is replaced with adipose
tissue. However, the exact effect of aging on thymic function remains
to be settled despite this evidence of morphological changes.
Although we cannot provide a definitive explanation for this
discrepancy, one possibility might be that lymphocytes with the Ph
chromosome fail to differentiate. This is compatible with the finding
that the B-cell population is chimeric with respect to the BCR/ABL and
that Ph+ T-cell progenitors may undergo only very limited
differentiation in the absence of an active thymus. Inducing the
Ph+ lymphoid progenitors to differentiate in vitro could
support our explanation, but the appropriate experimental procedures
and equipment are not available at this time.
In conclusion, mature T cells and NK cells in most CML patients are
Ph, but most patients have a mixture of
Ph+ and Ph B cells. Moreover, in all
patients with CML, BCR/ABL fusion was found in pluripotent stem cells,
B progenitor cells, T/NK progenitor cells, and T progenitor cells. The
application of FISH to blood and BM, smear FISH, yields important data
principally about mature cells. Furthermore, the technique of sorter
FISH (FACS + FISH) is a powerful tool to explore the cytogenetic
changes of immature cells of stem cell diseases. Further clarification
of genetic changes in stem cells could be achieved by using sorter FISH
with various combinations of monoclonal antibodies.
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FOOTNOTES |
Submitted April 14, 1998;
accepted July 16, 1998.
Address reprint request to Ikuo Miura, MD, Third Department of Internal
Medicine, Akita University School of Medicine, 1-1-1 Hondo, Akita,
Japan; e-mail: ikuo{at}med.akita-u.ac.jp.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
| |
REFERENCES |
1.
Fialkow PJ, Jacobson RJ, Papayannopoulou T:
Chronic myelocytic leukemia: Clonal origin in a stem cell common to the granulocyte, erythrocyte, platelet and monocyte/macrophage.
Am J Med
63:125, 1977[Medline]
[Order article via Infotrieve]
2.
Fialkow PJ, Denman AM, Jacobson RJ, Lowenthal MN:
Chronic myelocytic leukemia: Origin of some lymphocytes from leukemic stem cells.
J Clin Invest
62:815, 1978
3.
Kurzrock R, Gutterman JV, Talpaz M:
The molecular genetics of Philadelphia chromosome-positive leukemias.
N Engl J Med
319:990, 1988[Medline]
[Order article via Infotrieve]
4.
Rosenthal S, Canellos GP, DeVita VT, Gralnick HR:
Characteristics of blast crisis in chronic granulocytic leukemia.
Blood
49:705, 1977[Abstract/Free Full Text]
5.
Griffin J, Todd RF, Ritz J, Nadler LM, Canellos GP, Rosenthal D, Gallivan M, Beveridge PPS, Weinstein H, Karp D, Schlossman SF:
Differentiation patterns in the blastic phase of chronic myeloid leukemia.
Blood
61:85, 1983[Abstract/Free Full Text]
6.
Chan LC, Furley AJ, Ford AM, Yardumian DA, Greaves MF:
Clonal rearrangement and expression of T cell receptor  gene and involvement of the breakpoint cluster region in blast crisis of CGL.
Blood
67:533, 1986[Abstract/Free Full Text]
7.
Gramatzki M, Bartram CR, Muller D, Tittelbach M, Kalden JR:
Early T cell differentiated chronic myeloid leukemia blast crisis with rearrangement of the breakpoint cluster region but not of the T cell receptor chain genes.
Blood
69:1082, 1987[Abstract/Free Full Text]
8.
Lawler SD:
The cytogenetics of chronic granulocytic leukemia.
Clin Hematol
6:55, 1977
9.
Juneja HS, Weiner R:
Presence of the Philadelphia chromosome (Ph1) in pokeweed mitogen stimulated lymphocytes during chronic phase of chronic myelocytic leukemia (CML).
Cancer Genet Cytogenet
4:39, 1981[Medline]
[Order article via Infotrieve]
10.
Bagnara GP, Biagini G, Marani M, Bonsi L, Severi B, Valvassori L, Comis M, Nobile F, Iacopino P, Ronco F, Neri A:
Ph1-negative T lymphocytic colonies in agar cultures of peripheral blood in chronic myeloid leukemia.
Acta Haematol
66:174, 1981[Medline]
[Order article via Infotrieve]
11.
Kearney L, Orchard KH, Hibbin J, Goldman JM:
T-cell cytogenetics in chronic granulocytic leukemia.
Lancet
1:858, 1982[Medline]
[Order article via Infotrieve]
12.
Nitta M, Kato Y, Strife A, Wachter M, Fried J, Perez A, Jhanwar S, Duigou-Osterndorf R, Chaganti RSK, Clarkson B:
Incidence of involvement of B and T lymphocyte lineages in chronic myelogenous leukemia.
Blood
66:1053, 1985[Abstract/Free Full Text]
13.
Bartram CR, Raghavachar A, Anger B, Stain C, Bettelheim P:
T lymphocytes lack rearrangement of the bcr gene in Philadelphia chromosome-positive chronic myelocytic leukemia.
Blood
69:1682, 1987[Abstract/Free Full Text]
14.
Barr RD, Watt J:
Preliminary evidence for the common origin of a lympho-myeloid complex in man.
Acta Haematol
60:29, 1978[Medline]
[Order article via Infotrieve]
15.
Shabtai F, Gafter U, Weiss S, Djaldetti M, Halbrecht I:
New complex Ph translocation t(10;14;22) in bone marrow cells and in PHA-stimulated peripheral blood cultures in chronic myelocytic leukemia.
J Cancer Res Clin Oncol
96:287, 1980[Medline]
[Order article via Infotrieve]
16.
Torlakovic E, Litz CE, McClure JS, Brunning RD:
Direct detection of the Philadelphia chromosome in CD20-positive lymphocytes in chronic myeloid leukemia by tri-color immunophenotyping/FISH.
Leukemia
8:1940, 1994[Medline]
[Order article via Infotrieve]
17.
Haferlach T, Winkemann M, Nickenig C, Meeder M, Ramm-Petersen L, Schoch R, Nickelsen M, Weber-Matthiesen K, Schlegelberger B, Schoch C, Gassmann W, Löffler H:
Which compartments are involved in Philadelphia-chromosome positive chronic myeloid leukemia? An answer at the single cell level by combining May-Grünwald-Giemsa staining and fluorescence in situ hybridization techniques.
Br J Haematol
97:99, 1997[Medline]
[Order article via Infotrieve]
18.
Jonas D, Lubbert M, Kawasaki ES, Henke M, Bross KJ, Mertelsmann R, Herrmann F:
Clonal analysis of bcr-abl rearrangement in T lymphocytes from patients with chronic myelogenous leukemia.
Blood
79:1017, 1992[Abstract/Free Full Text]
19.
Fialkow PJ, Martin PJ, Najfeld V, Penfold GK, Jacobson RJ, Hansen JA:
Evidence for a multistep pathogenesis of chronic myelogenous leukemia.
Blood
58:158, 1981[Abstract/Free Full Text]
20.
Knuutila S, Teerenhovi L, Larramendy ML, Elonen E, Franssila KO, Nylund SJ, Timonen T, Heinonen K, Mahlamäki E, Winqvist R, Ruutu T:
Cell lineage involvement of recurrent chromosomal abnormalities in hematologic neoplasms.
Genes Chromosom Cancer
10:95, 1994[Medline]
[Order article via Infotrieve]
21.
Tkachuk DC, Westbrook CA, Andreeff M, Donlon TA, Cleary ML, Suryanarayan K, Homge M, Redner A, Gray J, Pinkel D:
Detection of bcr-abl fusion in chronic myelogenous leukemia by in situ hybridization.
Science
250:559, 1990[Abstract/Free Full Text]
22.
Craig W, Kay R, Cutler RL, Lansdorp PM:
Expression of Thy-1 on human hematopoietic progenitor cells.
J Exp Med
177:1331, 1993[Abstract/Free Full Text]
23.
Baum CM, Weissman IL, Tsukamoto AS, Buckle A-M, Peault B:
Isolation of a candidate human hematopoietic stem-cell population.
Proc Natl Acad USA
89:2804, 1992[Abstract/Free Full Text]
24.
Pontvert-Delucq S, Breton-Gorius J, Schmitt C, Ballou C, Guichard J, Najman A, Lemoine FM:
Characterization and functional analysis of adult human bone marrow cell subsets in relation to B-lymphoid development.
Blood
82:417, 1993[Abstract/Free Full Text]
25.
Spits H, Lanier LL, Phillips JH:
Development of human T and natural killer cells.
Blood
85:2654, 1995[Free Full Text]
26.
Miller JS, Alley KA, McGlave P:
Differentiation of natural killer (NK) cells from human primitive marrow progenitors in a stroma-based long-term culture system: Identification of a CD34+7+ NK progenitor.
Blood
83:2594, 1994[Abstract/Free Full Text]
27.
Barcena A, Muench MO, Galy AHM, Cupp J, Roncarolo MG, Phillips JH, Spits H:
Phenotypic and functional analysis of T-cell precursors in the human fetal liver and thymus: CD7 expression in the early stage of T- and myeloid-cell development.
Blood
82:3401, 1993[Abstract/Free Full Text]
28.
Bedi A, Zehnbauer BA, Collector MI, Barber JP, Zicha MS, Sharkis, Jones RJ:
BCR-ABL gene rearrangement and expression of primitive hematopoietic progenitors in chronic myeloid leukemia.
Blood
81:2898, 1993[Abstract/Free Full Text]
29.
Norman A, Sasaki MS, Ottoman RE, Fingerhut AG:
Lymphocyte lifetime in women.
Science
147:745, 1965[Abstract/Free Full Text]
30.
Buckton KE, Brown WMC, Smith PG:
Lymphocyte survival in men treated with X-ray for Ankylosing Spondylitis.
Nature
214:470, 1967[Medline]
[Order article via Infotrieve]
31.
Nanao K, Haruto U:
Chronologic sequence in appearance of clinical and laboratory findings characteristic of chronic myelocytic leukemia.
Blood
51:843, 1978[Free Full Text]
32.
Dreazen O, Canaani E, Gale RP:
Molecular biology of chronic myelogenous leukemia.
Semin Hematol
25:35, 1988[Medline]
[Order article via Infotrieve]
33.
Mrozek E, Anderson P, Caligiuri MA:
Rolle of interleukin-15 in development of human CD56+ natural killer cells from CD34+ hematopoietic progenitor cells.
Blood
87:2632, 1996[Abstract/Free Full Text]
34.
Miller JS, Verfaillie C, McGlave P:
The generation of human natural killer cells from CD34+/DR primitive progenitors in long-term bone marrow culture.
Blood
80:2182, 1992[Abstract/Free Full Text]
35.
Lotzova E, Savary CA, Champlin RE:
Genesis of human oncolytic natural killer cells from primitive CD34+CD33 bone marrow progenitor.
J Immunol
150:5263, 1993[Abstract]
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C. James, F. Mazurier, S. Dupont, R. Chaligne, I. Lamrissi-Garcia, M. Tulliez, E. Lippert, F.-X. Mahon, J.-M. Pasquet, G. Etienne, et al.
The hematopoietic stem cell compartment of JAK2V617F-positive myeloproliferative disorders is a reflection of disease heterogeneity
Blood,
September 15, 2008;
112(6):
2429 - 2438.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. W.N. Deininger
Imatinib Resistance and the Difficulty of Eradicating Leukemia Stem Cells
ASCO Educational Book,
January 1, 2008;
2008(1):
318 - 323.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Chaligne, C. James, C. Tonetti, R. Besancenot, J. P. Le Couedic, F. Fava, F. Mazurier, I. Godin, K. Maloum, F. Larbret, et al.
Evidence for MPL W515L/K mutations in hematopoietic stem cells in primitive myelofibrosis
Blood,
November 15, 2007;
110(10):
3735 - 3743.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. A. J. Signer, E. Montecino-Rodriguez, O. N. Witte, J. McLaughlin, and K. Dorshkind
Age-related defects in B lymphopoiesis underlie the myeloid dominance of adult leukemia
Blood,
September 15, 2007;
110(6):
1831 - 1839.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Delhommeau, S. Dupont, C. Tonetti, A. Masse, I. Godin, J.-P. L. Couedic, N. Debili, P. Saulnier, N. Casadevall, W. Vainchenker, et al.
Evidence that the JAK2 G1849T (V617F) mutation occurs in a lymphomyeloid progenitor in polycythemia vera and idiopathic myelofibrosis
Blood,
January 1, 2007;
109(1):
71 - 77.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. S. M. Yong, R. M. Szydlo, J. M. Goldman, J. F. Apperley, and J. V. Melo
Molecular profiling of CD34+ cells identifies low expression of CD7, along with high expression of proteinase 3 or elastase, as predictors of longer survival in patients with CML
Blood,
January 1, 2006;
107(1):
205 - 212.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Kosugi, Y. Ebihara, T. Nakahata, H. Saisho, S. Asano, and A. Tojo
CD34+CD7+ Leukemic Progenitor Cells May Be Involved in Maintenance and Clonal Evolution of Chronic Myeloid Leukemia
Clin. Cancer Res.,
January 15, 2005;
11(2):
505 - 511.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Yin, Y.-L. Wu, H.-P. Sun, G.-L. Sun, Y.-Z. Du, K.-K. Wang, J. Zhang, G.-Q. Chen, S.-J. Chen, and Z. Chen
Combined effects of As4S4 and imatinib on chronic myeloid leukemia cells and BCR-ABL oncoprotein
Blood,
December 15, 2004;
104(13):
4219 - 4225.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Streubel, A. Chott, D. Huber, M. Exner, U. Jager, O. Wagner, and I. Schwarzinger
Lymphoma-Specific Genetic Aberrations in Microvascular Endothelial Cells in B-Cell Lymphomas
N. Engl. J. Med.,
July 15, 2004;
351(3):
250 - 259.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Jaiswal, D. Traver, T. Miyamoto, K. Akashi, E. Lagasse, and I. L. Weissman
Expression of BCR/ABL and BCL-2 in myeloid progenitors leads to myeloid leukemias
PNAS,
August 19, 2003;
100(17):
10002 - 10007.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Dong, K. Cwynarski, A. Entwistle, F. Marelli-Berg, F. Dazzi, E. Simpson, J. M. Goldman, J. V. Melo, R. I. Lechler, I. Bellantuono, et al.
Dendritic cells from CML patients have altered actin organization, reduced antigen processing, and impaired migration
Blood,
May 1, 2003;
101(9):
3560 - 3567.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. S. Yavuz, P. E. Lipsky, S. Yavuz, D. D. Metcalfe, and C. Akin
Evidence for the involvement of a hematopoietic progenitor cell in systemic mastocytosis from single-cell analysis of mutations in the c-kit gene
Blood,
June 28, 2002;
100(2):
661 - 665.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Nakajima, R. Zhao, T. C. Lund, J. Ward, M. Dolan, B. Hirsch, and J. S. Miller
The BCR/ABL Transgene Causes Abnormal NK Cell Differentiation and Can Be Found in Circulating NK Cells of Advanced Phase Chronic Myelogenous Leukemia Patients
J. Immunol.,
January 15, 2002;
168(2):
643 - 650.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. H. S. Kabarowski and O. N. Witte
Consequences of BCR-ABL Expression within the Hematopoietic Stem Cell in Chronic Myeloid Leukemia
Stem Cells,
November 1, 2000;
18(6):
399 - 408.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
L. Nilsson, I. Astrand-Grundstrom, I. Arvidsson, B. Jacobsson, E. Hellstrom-Lindberg, R. Hast, and S. E. W. Jacobsen
Isolation and characterization of hematopoietic progenitor/stem cells in 5q-deleted myelodysplastic syndromes: evidence for involvement at the hematopoietic stem cell level
Blood,
September 15, 2000;
96(6):
2012 - 2021.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Kosugi, A. Tojo, H. Shinzaki, T. Nagamura-Inoue, and S. Asano
The preferential expression of CD7 and CD34 in myeloid blast crisis in chronic myeloid leukemia
Blood,
March 15, 2000;
95(6):
2188 - 2189.
[Full Text]
[PDF]
|
|
|
|
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Top
Abstract
Introduction
)
and monocytes (
).
