|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 95 No. 2 (January 15), 2000:
pp. 404-408
CLINICAL OBSERVATIONS, INTERVENTIONS, AND THERAPEUTIC TRIALS
Persistence of BCR-ABL genomic rearrangement in chronic myeloid
leukemia patients in complete and sustained cytogenetic remission after
interferon- therapy or allogeneic bone marrow transplantation
Jean-Claude Chomel,
Françoise Brizard,
Anne Veinstein,
Jérôme Rivet,
Alain Sadoun,
Alain Kitzis,
François Guilhot, and
André Brizard
From the Laboratoire de Génétique Cellulaire et
Moléculaire (UPRES EA 2622), the Laboratoire d'Hématologie
(CNRS ESA 6031), and the Département d'Hématologie et
Oncologie Médicale (UPRES EA 2622), CHU de Poitiers, France.
 |
Abstract |
In recent years, the prognosis of chronic myeloid leukemia (CML) has
been greatly improved either with interferon- (IFN-) therapy or
allogeneic bone marrow transplantation (BMT). In the present study,
minimal residual disease was evaluated in 21 patients in complete
cytogenetic response (CCR) after such treatments. Samples from bone
marrow aspirates or peripheral blood or both were analyzed by
conventional cytogenetics, Southern blot, interphase fluorescent in
situ hybridization (FISH), and quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR). In all patients, FISH detected 1% to 12% nuclei with a BCR-ABL fusion gene, whereas Q-RT-PCR experiments were negative or weakly positive. Based on these
results, we hypothesize that the BCR-ABL genomic rearrangement persists
unexpressed in nonproliferating cells whatever the treatment (IFN-
or BMT). These data point to the need for follow-up of CML patients in
CCR over an extensive period at the DNA level (FISH) to evaluate the
residual disease and at the RNA level (Q-RT-PCR) to estimate the risk
of relapse.
(Blood. 2000;95:404-408)
© 2000 by The American Society of Hematology.
| |
Introduction |
Chronic myeloid leukemia (CML) is characterized in more
than 90% of patients by a reciprocal translocation between chromosomes 9 and 22: translocation t(9;22)(q34;q11), which is referred to as the
Philadelphia chromosome (Ph).1,2 At the molecular level,
the Ph chromosome results from a rearrangement between the ABL gene
located on chromosome 9 and the BCR gene on chromosome 22 forming a
BCR-ABL hybrid gene, which is a specific marker of the
disease.3,4 In recent years, the prognosis of CML has been
improved by allogeneic bone marrow transplantation (BMT) or
interferon- (IFN-) therapy. It has been shown that CML can be
cured by allografting and that IFN- (with or without cytarabine) is
able to induce a complete cytogenetic response in up to 26% of
patients treated at diagnosis.5 In such cases, the question of maintenance therapy (dose and duration) remains unresolved. Hence,
the definition of complete remission must be as accurate as possible.
In a preliminary study,6 we used cytogenetics, interphase
fluorescent in situ hybridization (FISH), and quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) methods to study
minimal residual disease in 3 patients with CML in sustained complete
cytogenetic remission after treatment with IFN-. The BCR-ABL fusion
gene was still detectable by FISH, whereas RT-PCR remained negative or
weakly positive. However, the precision of the percentage of
BCR-ABL-positive nuclei was hampered by the rather high percentage of
false-positive nuclei due to the use of BCR-ABL DNA probes that
detected only a single BCR-ABL fusion signal. New FISH probes (D-FISH
probes) are now available, and they detect BCR-ABL fusion in interphase
nuclei with a false-positive signal rate close to zero.7,8
Such probes were used in the current study, which was performed in 21 CML patients in sustained complete cytogenetic remission, either after
IFN- therapy (11 cases) or allogeneic BMT (10 cases). To monitor the
minimal residual disease, conventional cytogenetics, Southern blot,
FISH, and Q-RT-PCR were performed. Persistence of the BCR-ABL genomic
rearrangement was evidenced by FISH in all cases, whereas most Q-RT-PCR
experiments remained negative or weakly positive.
| |
Patients and methods |
Patients
Cells from 21 consecutive CML patients (10 men and 11 women) were
studied. At presentation, 17 patients presented the classical t(9;22)(q34;q11) translocation (additional abnormalities were detected
in a single case) and 4 patients had a complex chromosomal rearrangement (Tables 1 and
2). The molecular rearrangement in M-BCR
(major-breakpoint cluster region) was attested by Southern blot
analysis. The type of chimeric transcript was assessed by qualitative
RT-PCR experiments; all patients had the typical b2a2 or b3a2
transcript. After treatment with IFN- or allogeneic BMT, the
monitoring of the residual disease mainly involved cytogenetic analyses. When the patient had reached a partial or complete
cytogenetic response, FISH, Southern blot (in the majority of cases),
and Q-RT-PCR were performed together with further cytogenetic analyses. Informed consent was obtained as required by the Declaration of Helsinki.
Eleven patients (7 men and 4 women, median age at diagnosis 53 years,
range 27-70 years) were studied at the time of diagnosis and during
IFN- therapy (see Table 1). All were in chronic phase at
presentation, most with a low (6 cases) or intermediate score (4 cases)
according to Sokal's index.9 These patients were treated,
using various doses of hydroxyurea, IFN-, and cytarabine, based on
data from 3 consecutive trials.10-12 A cytogenetic
evaluation was performed every 3 or 4 months for the first 12 months
and then at 4- or 6-month intervals. All patients achieved a complete cytogenetic response (CCR) after a median treatment period of 24 months
(range, 5-40 months). The median follow-up of these patients is 127 months (range, 41-183 months). Currently, all patients are alive and in
persistent CCR.
Ten other patients (3 men and 7 women, median age at diagnosis 31 years, range 13-48 years) received an allogeneic non-T-cell-depleted bone marrow transplant from an HLA-identical sibling donor (cases 12-14, 16-19, and 21) or matched unrelated donor (cases 15 and 20)
after a median period of treatment with IFN- or chemotherapy (mainly
hydroxyurea) or both of 18 months since diagnosis (range, 3-57) (see
Table 2). Patients 13 and 19 received a sex-mismatched allogeneic BMT.
Most patients had at diagnosis a moderate Gratwohl's score.13 All patients were in first chronic phase, and most of them underwent transplantation because of treatment failure with no
cytogenetic response. Patient 13, who had massive splenomegaly, underwent splenectomy before transplant. The conditioning regimen was
either total body irradiation and cyclophosphamide or busulfan and
cyclophosphamide. All patients received methotrexate (4 days) and
cyclosporine as prophylaxis for graft-versus-host disease. The median
follow-up time from diagnosis for the 10 BMT patients is 47 months
(range, 29-95 months). All patients had normal karyotypes after
grafting. Currently, patients 12, 13, 14, 17, 18, and 21 are well;
patients 15, 19, and 20 are affected by a chronic graft-versus-host disease; and patient 16 is well after a donor lymphocyte infusion.
Cytogenetic and FISH analysis
Bone marrow aspirates or peripheral blood samples or both were
obtained for simultaneous cytogenetic and molecular studies. Chromosome
analysis was performed according to the R-banding method after short
(24- or 48-hour) cultures without addition of mitogens. FISH analysis
was performed on unseparated nucleated cells from cytogenetic
preparations or fresh blood samples, using differently colored,
directly labeled BCR and ABL probes developed by ONCOR (D-FISH,
Gaithersburg, MD) according to manufacturer's instructions. A minimum
of 200 cells in interphase were scored for each sample. Normal cells
display 2 red signals and 2 green signals (2R2G), and abnormal cells 1 red, 1 green, and 2 yellow fusion signals (1R1G2F) or 2 red, 2 green,
and 1 yellow fusion signals (2R2G1F) or 1 red, 1 green, and 3 yellow
fusion signals (1R1G3F) indicating 2 Ph chromosomes.7,8 The
positivity pattern for each specimen was recorded and final results
were expressed as percentages of nuclei with fusion signals.
Negative FISH control studies were performed with blood or bone marrow
smears from 5 patients with hematologic disorders other than CML. A
total of 1000 interphase nuclei were scored and a single nucleus with a
positive pattern (2R2G1F) was found. The cutoff limit for BCR-ABL
positivity was hence set at the mean of normal result (0.1% + 3 SD,
i.e., 0.3%). Positive controls were also performed with bone marrow
cultures or uncultured blood smears from 6 patients in chronic phase of
CML with a percentage of Ph+ metaphases ranging from 50% to 100%. By
FISH, the percentage of positive interphase cells was similar to that
of positive metaphases (range, 57-100%).
In the 2 sex-mismatched BMT cases (patients 13 and 19), the chimerism
was studied by FISH using specific Y and X chromosome probes: LSI SRY
probe (VYSIS Inc, Downers Grove, IL) and chromosome X alpha satellite
probe (DXZ1, ONCOR, Gaithersburg, MD), respectively.
Southern blot
DNA for Southern blot analysis was available from patients 1 through
6, 8 through 11, 13, 14, and 20. DNA (15 µg) was digested with BglII
restriction enzyme, electrophoresed on a 0.8% agarose gel, and
transferred onto a nylon filter. After 32P-labeling, the
phl/bcr-3 fragment (Oncogene Science, Uniondale, NY) was used
as a probe. In the absence of the BCR-ABL translocation at the M-BCR
locus, 3 DNA fragments were visualized. When a translocation had
occurred, 1 or 2 additional fragments were present. To compare results
from FISH analysis and Southern blot, mixtures of normal and 100% Ph+
cell samples at different ratios were prepared (Table 3). Rearranged fragments observed by
Southern blot were semiquantified by densitometry. In every dilution,
the percentages of Ph+ metaphases and positive nuclei by FISH were
similar. For the same dilutions, the percentage of rearranged fragments
appreciated by densitometry after Southern blot was approximately half
the percentage of Ph+ cells. A pure leukemic cell population has 1 normal chromosome 22 and 1 Ph+ chromosome. Every cell in such a
population is positive by FISH (100% of positive nuclei), whereas the
percentage of rearranged DNA fragments observed by Southern blot is
theoretically close to 50%.14 Therefore, results from
these 2 methods are not directly equivalent, the first yielding a
percentage of nuclei with at least 1 BCR-ABL rearrangement, the other
allowing an estimation of the rearrangement in M-BCR at the DNA level.
Qualitative and quantitative RT-PCR
Total leukocyte RNA was extracted from the peripheral blood because
Q-RT-PCR for the monitoring of residual disease in CML yields similar
results on blood or bone marrow samples.15 RNA was
extracted according to Chomczynski and Sacchi16 and
controlled by electrophoresis on 1% agarose gel. Reverse transcription
and qualitative and quantitative RT-PCR for ABL-mRNA and p210
BCR-ABL-mRNA were performed according to a standardized protocol using
serial dilutions of a p210 competitor ranging from 1 to 107
molecules.15,17 Results were expressed as the
BCR-ABL/ABL mRNA ratio. In our hands, the method is able to detect as
few as 3 BCR-ABL mRNA molecules in the assay. On average, 150,000 ABL
transcript molecules per assay are found. Therefore, the sensitivity of
the amplification can be estimated by a BCR-ABL/ABL ratio close to
0.002% (3/150,000). In this study, a RT-PCR is considered as negative
when the BCR-ABL/ABL ratio is equal to or lower than 0.002%.
Quantitative RT-PCR was compared with cytogenetic findings in 104 CML
cases treated with IFN- in our institution (including the 11 patients in CCR of the present study). Cytogenetic responders were
considered as complete responders (Ph+ = 0%), partial responders (0% < Ph+  34%), minor responders (34% < Ph+ 94%), and
nonresponders (Ph+ > 94%), with median BCR-ABL/ABL ratios of 0.01%,
2%, 14%, and 34%, respectively (Figure
1). According to these results, we defined
2 arbitrary values (1% and 0.01%) for the BCR-ABL/ABL ratio. A ratio
below 1% is generally associated with a CCR and a ratio below 0.01%
with a long and stable CCR.
View larger version (14K):
[in this window]
[in a new window]
| Fig 1.
Comparison of the BCR-ABL/ABL ratio with the cytogenetic
response status.
Results include 104 CML cases treated with IFN- in our institution
(including the 11 patients in CCR of the present study).
|
|
| |
Results |
Cytogenetic analysis
At the time of FISH study, 10 of the 11 patients treated with
IFN- had been in CCR for 2 to 10 years. The last patient (no. 5)
achieved a complete response 1 year before FISH study, but very few
positive metaphases (18/642 Ph+ metaphases in 7/22 samples) had been
found during the 9 previous years of follow-up. At the time of study,
karyotypic analysis failed to detect any Ph+ metaphases in the samples
from the 10 patients who received BMT.
FISH and Southern blot experiments
In all 21 patients, FISH analyses detected a BCR-ABL fusion gene
(Table 4). The percentage of fusion signals
ranged from 1% to 12% nuclei (median 6-7%). Patients 1, 2, and 3, with 1% to 2% positive nuclei by FISH, were analyzed at 3 consecutive times with identical results. There was no significant relationship between percentages of positive nuclei and duration of CCR or BCR-ABL/ABL ratios. However, patients 1 and 2 (CCR > 9 years) showed
the lowest percentages of both positive nuclei by FISH (2% and 1%,
respectively) and BCR-ABL/ABL ratio (< 0.002%).
In 2 BMT cases, donor and recipient were of different sex (patients 13 and 19). Because of the presence of a significant percentage of BCR-ABL
rearranged cells in CML patients in CCR after BMT, the presence of
residual host cells harboring the BCR-ABL rearrangement was suspected.
To test this hypothesis, FISH with X and Y probes was performed from
the same samples as those studied by FISH with BCR and ABL probes. In
patient 19 (female recipient and male donor), 3% of XX host cells were
detected as compared with 3% of cells with BCR-ABL fusion signals. In
patient 13 (male recipient, female donor), 4% of XY-positive nuclei
were detected as compared with 6% of cells with BCR-ABL fusion
signals. Moreover, 2 years before the present study, Y chromosome
sequences were detected by PCR18 in patient 13, and FISH
analyses on the same samples revealed 6% of BCR-ABL rearranged cells
and 5% of XY nuclei.
Southern blot analyses were negative in patients 1, 2, 3, 8, 11, and
13, but showed weak rearranged bands (< 5% of rearranged DNA
fragments) in patients 4, 6, 9, 10, 14, and 20. In patient 5, in whom a
CCR occurred only during the last year preceding FISH, rearranged bands
reached a level of about 5% to 10%. Such discrepancies between FISH
and Southern blot results can be explained by a number of rearranged
cells of about 6% to 7%, which corresponds to a percentage of
rearranged DNA fragments close to the detection limit of the Southern
blot method.
Evolution of the BCR-ABL/ABL ratio
The kinetics of the BCR-ABL/ABL ratio was evaluated by Q-RT-PCR
during the last 2 years of the study. In patients 1, 2, 3, and 7 (IFN- therapy) and patients 12 through 15 and 17 through 21 (BMT),
BCR-ABL/ABL ratios were stable and remained below 0.01% (Figure
2). These ratios fluctuated close to 0.01%
for patients 6 and 8 and varied between 0.01% and 1% for patients 4, 5, 9, 10, and 11. In summary, most patients who received BMT had
persistent BCR-ABL/ABL ratios below 0.01% (FISH median 7%). Among the
CML patients treated with IFN-, 6 showed similar kinetics, with
BCR-ABL/ABL ratios below or close to 0.01% (FISH median 2%) and 5 had
ratios above this value (FISH median 7%). In patient 16, a molecular relapse occurred after 2 years of CCR and was followed by a cytogenetic relapse 2 months later. Infusion of the BMT donor's lymphocytes resulted in a second CCR with BCR-ABL/ABL ratio below 0.01% and FISH
at 6% (see Figure 2).
View larger version (27K):
[in this window]
[in a new window]
| Fig 2.
Evolution of the BCR-ABL/ABL ratio in CML patients
treated with IFN- (patients 1-10) or BMT (patient 16).
The BCR-ABL/ABL ratio kinetics for patient 16 is characteristic of a
transient relapse. Other patients in CCR after BMT are not shown
because they usually have BCR-ABL/ABL ratios close to 0.002%.
|
|
| |
Discussion |
In a preliminary study of cells from 3 CML patients treated with
IFN- in long CCR,6 we observed the persistence of
BCR-ABL rearranged cells detected by FISH, whereas conventional
cytogenetics was negative and Q-RT-PCR negative or weakly positive. We
concluded that the malignant cells harboring the genomic BCR-ABL
rearrangement were nonproliferating neoplastic cells. These cells did
not express significant levels of BCR-ABL transcript and a
fortiori did not synthesize the chimeric p210BCR-ABL
protein. In the present study, we extended our preliminary data by
using cytogenetics, Southern blot, FISH, and Q-RT-PCR methods to 21 patients in sustained CCR after treatment with IFN- or BMT.
To accurately monitor the residual disease in CML, the methods must be
well standardized, especially for FISH and Q-RT-PCR methods. In our
preliminary study,6 the level of false-positive results by
FISH reached 3% to 6%, whereas in the present study, using more
specific BCR and ABL probes, they were close to 0.1%. Comparison
between FISH and Southern blot showed that FISH is more appropriate for
the quantification of small percentages (1-5%) of rearranged cells
than is Southern blot, in accordance with previous data showing that a
linear relationship between the percentages of rearranged fragments and
of Ph+ cells is observed only in the range 20% to 100% of Ph+
cells.14 Moreover, weak bands on autoradiographs could not
be correctly quantified. When the BCR-ABL/ABL ratio determined by
Q-RT-PCR was compared with the cytogenetic response status of the
patient, the results were similar to those published by Hochhaus et
al.15 These results allowed us to define 2 cutoff points of
1% and 0.01%. Most patients in CCR have a BCR-ABL/ABL ratio below
1%, and most patients in long and stable CCR have a BCR-ABL/ABL ratio
below 0.01%. It is thus important to underline the peculiar objective
of cytogenetics, FISH, Southern blot, and Q-RT-PCR for the monitoring
of patients with CML. (1) Conventional cytogenetics analyzes cycling
cells, with results being expressed as the percentage of metaphases
harboring the translocation t(9;22). (2) FISH methods test every cell
with results in percentages of positive nuclei harboring the
BCR-ABL genomic rearrangement. (3) Southern blot detects M-BCR
rearrangements at the DNA level, and results are given in a
boolean manner (presence or absence of rearranged DNA fragments) with a
percentage of rearranged DNA estimated by densitometry. (4) Q-RT-PCR
does not quantify the percentage of malignant cells, but BCR-ABL fusion
mRNA and the results are expressed by the BCR-ABL/ABL ratio. Therefore,
these methods are not equivalent but complementary.
In the patients in long CCR after IFN- therapy, we confirm the
presence of rearranged cells (median 6-7%) in the absence of
significant levels of BCR-ABL transcripts. This result is not related
to a lack of sensitivity of Q-RT-PCR experiments because, in our hands,
this method can detect as few as 3 BCR-ABL mRNA molecules in the assay.
It is noteworthy that, in our experience, we have never seen CML
patients in CCR with D-FISH negative. However, the percentage of
FISH-positive cells seems to be weaker in patients in very long CCR.
Such apparent discrepancies between cytogenetics or FISH or Southern
blot and RT-PCR have been already mentioned.19-26 In most
of these reports, studies were performed after cultures of
hematopoietic progenitor cells from untreated patients with CML,19-22,26 and it has been suggested that the BCR-ABL
translocation was present in the most primitive hematopoietic
progenitor cells, which might have been quiescent and transcriptionally
silent. In our previous study, we postulated that this observation
could be the consequence of the molecular action of IFN-. Actually, IFN- exerts various activities on cellular proliferation and immunoregulation.27,28 However, FISH analysis of the cells of the 10 patients in CCR after BMT showed similar results as after
IFN- therapy (6-7% of rearranged cells). In the 2 sex-mismatched BMT, we showed that a significant minority (3-6%) of hematopoietic cells are of host origin. These findings suggest that the persistence of nonproliferating neoplastic cells is a general feature of CML patients in CCR whatever the treatment. FISH has been performed from
unseparated blood or bone marrow nucleated cells, and the nature of the
FISH-positive cells could not be easily determined. One could speculate
that some of the FISH-positive cells could represent residual
lymphocytes in the IFN- responders, but this explanation is
improbable in the patients after BMT. The reasons for this persistence
state and for the nonexpansion of the malignant clone are unclear. An
early and transient relapse occurred in 1 of the 21 patients studied
(patient 16 treated with BMT). It remains possible that the rearranged
cells might re-enter the cell cycle, leading to the expansion of the
malignant clone. In this situation, RT-PCR follow-up may be a precious
help to predict a relapse.29 For example, in a patient not
included in this study, we detected a molecular relapse by Q-RT-PCR in
the absence of Ph+ metaphase and 7% positive nuclei by FISH. This
patient was previously in CCR after allogeneic BMT; Q-RT-PCR was
negative at 3 consecutive times during 7 months of follow-up, until the occurrence of a sudden and persistent increase of the BCR-ABL/ABL ratio
(10%, then 50%) that preceded cytogenetic relapse and an acute
myeloid transformation.
In summary, a significant number of nonproliferating neoplastic cells
persist in patients with CML whatever the treatment (IFN- or BMT).
These data would provide a rational explanation for the relapses
observed after allografting. However, a substantial number of patients
treated by IFN- and most patients who received BMT do not relapse
over an extensive period. This suggests that the residual
BCR-ABL-positive cells detected by FISH in CML patients in CCR may be
dormant according to the definition of Uhr et al.30 Their
oncogenic potential could be then controlled by various mechanisms such
as specific immune response.31-33 Because the nature of the
nonproliferating BCR-ABL cells attested by FISH is still unknown, a
potential risk of relapse exists for patients with CML as long as they
have these rearranged hematopoietic cells. These data indicate the need
for a follow-up of patients with CML in CCR over an extensive period at
the DNA level (FISH) to evaluate the residual disease and at the RNA
level (Q-RT-PCR) to estimate the risk of relapse.
| |
Acknowledgment |
The authors are grateful to Pr Jean Louis Preud'homme for helpful comments.
| |
Footnotes |
Submitted June 9, 1999; accepted September 21, 1999.
Supported by the Association pour la Recherche contre le Cancer and the
Ligue Nationale contre le Cancer (comité de Charente-Maritime).
Reprints: Alain Kitzis, Laboratoire de Génétique
Cellulaire et Moléculaire, CHU de Poitiers, BP 577, 86021 Poitiers Cedex, France.
The publication costs of this
article were defrayed in part by
page charge payment. Therefore,
and solely to indicate this fact,
this article is hereby marked
"advertisement"
in accordance with 18 U.S.C.
section 1734.
| |
References |
1.
Nowell PC, Hungerford DA.
A minute chromosome in human chronic granulocytic leukemia.
Science.
1960;132:1497-1501.
2.
Rowley JD.
A new consistent chromosomal abnormality in chronic myelogenous leukemia identified by quinacrine fluorescence and Giemsa staining.
Nature.
1973;243:290-293[Medline]
[Order article via Infotrieve].
3.
Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G.
Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22.
Cell.
1984;36:93-99[Medline]
[Order article via Infotrieve].
4.
Shtivelman E, Lifshitz B, Gale RP, Canaani E.
Fused transcript of abl and bcr genes in chronic myelogenous leukaemia.
Nature.
1985;315:550-554[Medline]
[Order article via Infotrieve].
5.
Kantarjian HM, O'Brien S, Anderlini P, Talpaz M.
Treatment of chronic myelogenous leukemia: current status and investigational options.
Blood.
1996;87:3069-3081[Free Full Text].
6.
Brizard F, Chomel JC, Veinstein A, et al.
Does BCR-ABL genomic rearrangement persist in CML patients in complete remission after interferon therapy?
Leukemia.
1998;12:1076-1080[Medline]
[Order article via Infotrieve].
7.
Buno I, Wyatt WA, Zinsmeister AR, Dietz-Band J, Silver RT, Dewald GW.
A special fluorescent in situ hybridization technique to study peripheral blood and assess the effectiveness of interferon therapy in chronic myelogenous leukemia.
Blood.
1998;92:2315-2321[Abstract/Free Full Text].
8.
Dewald GW, Wyatt WA, Juneau AL, et al.
Highly sensitive fluorescence in situ hybridization method to detect double BCR/ABL fusion and monitor response to therapy in chronic myelogenous leukemia.
Blood.
1998;91:3357-3365[Abstract/Free Full Text].
9.
Sokal JE, Cox EB, Baccarani M, et al.
Prognostic discrimination in "good-risk" chronic granulocytic leukemia.
Blood.
1984;63:789-799[Abstract/Free Full Text].
10.
Guilhot F, Chastang C, Michallet M, et al.
(for the French Chronic Myeloid Leukemia Study Group). Interferon alfa-2b combined with cytarabine versus interferon alone in chronic myelogenous leukemia.
N Engl J Med.
1997;337:223-229[Abstract/Free Full Text].
11.
Guilhot F, Dreyfus B, Brizard A, Huret JL, Tanzer J.
Cytogenetic remissions in chronic myelogenous leukemia using interferon alpha-2a and hydroxyurea with or without low-dose cytosine arabinoside.
Leuk Lymphoma.
1991;4:49-55.
12.
Guilhot F, Guerci A, Fière D, et al.
The treatment of chronic myelogenous leukemia by interferon and cytosine-arabinoside: rationale and design of the French trials.
Bone Marrow Transplant.
1996;17(suppl 3):S29-S31.
13.
Gratwohl A, Hermans J, Goldman JM, et al.
(for the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation). Risk assessment for patients with chronic myeloid leukaemia before allogeneic blood or marrow transplantation.
Lancet.
1998;352:1087-1092[Medline]
[Order article via Infotrieve].
14.
Grossman A, Silver RT, Szatroswki TP, Gutfriend A, Verma RS, Benn PA.
Densitometric analysis of Southern blot autoradiographs and its application to monitoring patients with chronic myeloid leukemia.
Leukemia.
1991;5:540-547[Medline]
[Order article via Infotrieve].
15.
Hochhaus A, Lin F, Reiter A, et al.
Quantification of residual disease in chronic myelogenous leukemia patients on interferon- therapy by competitive polymerase chain reaction.
Blood.
1996;87:1549-1555[Abstract/Free Full Text].
16.
Chomczynski P, Sacchi N.
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
Anal Biochem
1987;162:156-159[Medline]
[Order article via Infotrieve].
17.
Cross NCP, Feng L, Chase A, Bungey J, Hughes TP, Goldman JM.
Competitive polymerase chain reaction to estimate the number of BCR-ABL transcripts in chronic myeloid leukemia patients after bone marrow transplantation.
Blood.
1993;82:1929-1936[Abstract/Free Full Text].
18.
Patri S, Dascalescu C, Chomel JC, et al.
Monitoring and prognostic evaluation of sex-mismatched bone marrow transplantation by competitive PCR on Y chromosome sequences.
Bone Marrow Transplant.
1996;17:625-632[Medline]
[Order article via Infotrieve].
19.
Kirk JA, Reems JA, Roecklein BA, et al.
Benign marrow progenitors are enriched in the CD34+/HLA-DR1o population but not in the CD34+/CD381o population in chronic myeloid leukemia: an analysis using interphase fluorescence in situ hybridization.
Blood.
1995;86:737-743[Abstract/Free Full Text].
20.
Verfaillie CM, Bhatia R, Miller W, et al.
BCR/ABL-negative primitive progenitors suitable for transplantation can be selected from the marrow of most early-chronic phase but not accelerated-phase chronic myelogenous leukemia patients.
Blood.
1996;87:4770-4779[Abstract/Free Full Text].
21.
Bedi A, Zehnbauer BA, Collector MI, et al.
BCR-ABL gene rearrangement and expression of primitive hematopoietic progenitors in chronic myeloid leukemia.
Blood.
1993;81:2898-2902[Abstract/Free Full Text].
22.
Leemhuis T, Leibowitz D, Cox G, et al.
Identification of BCR/ABL-negative primitive hematopoietic progenitor cells within chronic myeloid leukemia marrow.
Blood.
1993;81:801-807[Abstract/Free Full Text].
23.
Keating A, Wang XH, Laraya P.
Variable transcription of BCR-ABL by Ph+ cells arising from hematopoietic progenitors in chronic myeloid leukemia.
Blood.
1994;83:1744-1749[Abstract/Free Full Text].
24.
Yin JL, Williams BG, Arthur CK, Ma DDF.
Interferon response in chronic myeloid leukaemia correlated with ABL/BCR expression: a preliminary study.
Br J Haematol.
1995;89:539-545[Medline]
[Order article via Infotrieve].
25.
El-Rifai W, Ruutu T, Vettenranta K, Temtamy S, Knuutila S.
Minimal residual disease after allogeneic bone marrow transplantation for chronic myeloid leukaemia: a metaphase-FISH study.
Br J Haematol.
1996;92:365-369[Medline]
[Order article via Infotrieve].
26.
Maguer-Satta V, Petzer AL, Eaves AC, Eaves CJ.
BCR-ABL expression in different subpopulations of functionally characterized Ph+ CD34+ cells from patients with chronic myeloid leukemia.
Blood.
1996;88:1796-1802[Abstract/Free Full Text].
27.
Lengyel P.
Tumor-suppressor genes: news about the interferon connection.
Proc Natl Acad Sci U S A.
1993;90:5893-5895[Abstract/Free Full Text].
28.
Shirodkar S, Ewen M, DeCaprio JA, Morgan J, Livingston DM, Chitteriden T.
The transcription factor E2F interacts with the retinoblastoma product and a p107-cyclin A complex in a cell cycle-regulated manner.
Cell
1992;68:157-166[Medline]
[Order article via Infotrieve].
29.
Faderl S, Talpaz T, Kantarjian HM, Estrov Z.
Should polymerase chain reaction analysis to detect minimal residual disease in patients with chronic myelogenous leukemia be used in clinical decision making?
Blood.
1999;93:2755-2759[Free Full Text].
30.
Uhr JW, Scheuermann RH, Street N, Vitetta ES.
Cancer dormancy: opportunities for new therapeutic approaches.
Nat Med.
1997;3:505-509[Medline]
[Order article via Infotrieve].
31.
Goldman JM, Gale RP, Horowitz MM, et al.
Bone marrow transplantation for chronic myelogenous leukemia in chronic phase: increased risk for relapse with T-cell depletion.
Ann Intern Med.
1988;108:806-814.
32.
Sacchi S, Kantarjian H, Cohen P, Pierce S, Talpaz M.
Immune-mediated and unusual complications during alpha-interferon therapy in chronic myelogenous leukemia.
J Clin Oncol.
1995;13:2401-2407[Abstract/Free Full Text].
33.
Oka T, Sastry KJ, Nehete P, et al.
Evidence for specific immune response against P210 BCR-ABL in long-term remission CML patients treated with interferon.
Leukemia.
1998;12:155-163[Medline]
[Order article via Infotrieve].
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
Related Article in Blood Online:
-
No evidence for persistence of BCR-ABL-positive cells in patients in molecular remission after conventional allogeneic transplantation for chronic myeloid leukemia
- Michael Deininger, Tornalf Lehmann, Rainer Krahl, Eveline Hennig, Christel Müller, Dietger Niederwieser;, Françoise Brizard, Andre Brizard, Françoise Guilhot, Jean-Claude Chomel, and Alain Kitzis
Blood 2000 96: 778-780.
[Full Text]
[PDF]
Related Letter in Blood Online:
-
Absence of host-derived cells in the blood of patients in remission after allografting for chronic myeloid leukemia
- Andrew Chase, Sally Parker, Jaspal Kaeda, Renuka Sivalingam, Nicholas C. P. Cross, and John M. Goldman
Blood 2000 96: 777-778.
[Full Text]
[PDF]
This article has been cited by other articles:
|
|
|
|
 
M. Copland, A. Hamilton, L. J. Elrick, J. W. Baird, E. K. Allan, N. Jordanides, M. Barow, J. C. Mountford, and T. L. Holyoake
Dasatinib (BMS-354825) targets an earlier progenitor population than imatinib in primary CML but does not eliminate the quiescent fraction
Blood,
June 1, 2006;
107(11):
4532 - 4539.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. H.M. Jamieson, L. E. Ailles, S. J. Dylla, M. Muijtjens, C. Jones, J. L. Zehnder, J. Gotlib, K. Li, M. G. Manz, A. Keating, et al.
Granulocyte-Macrophage Progenitors as Candidate Leukemic Stem Cells in Blast-Crisis CML
N. Engl. J. Med.,
August 12, 2004;
351(7):
657 - 667.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Rosti, G. Martinelli, S. Bassi, M. Amabile, E. Trabacchi, B. Giannini, D. Cilloni, B. Izzo, A. De Vivo, N. Testoni, et al.
Molecular response to imatinib in late chronic-phase chronic myeloid leukemia
Blood,
March 15, 2004;
103(6):
2284 - 2290.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. M. Goldman and J. V. Melo
Chronic Myeloid Leukemia -- Advances in Biology and New Approaches to Treatment
N. Engl. J. Med.,
October 9, 2003;
349(15):
1451 - 1464.
[Full Text]
[PDF]
|
|
|
|
|
|
|
W.-I. Lee, H. Kantarjian, A. Glassman, M. Talpaz, and M.-S. Lee
Quantitative measurement of BCR/abl transcripts using real-time polymerase chain reaction
Ann. Onc.,
May 1, 2002;
13(5):
781 - 788.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Bagg
Chronic Myeloid Leukemia: A Minimalistic View of Post-Therapeutic Monitoring
J. Mol. Diagn.,
February 1, 2002;
4(1):
1 - 10.
[Full Text]
[PDF]
|
|
|
|
Top
Abstract
Introduction