Immature CD34+CD19− progenitor/stem cells in TEL/AML1-positive acute lymphoblastic leukemia are genetically and functionally normal

Blood ◽  
2002 ◽  
Vol 100 (2) ◽  
pp. 640-646 ◽  
Author(s):  
Marc Hotfilder ◽  
Silja Röttgers ◽  
Annegret Rosemann ◽  
Heribert Jürgens ◽  
Jochen Harbott ◽  
...  
Keyword(s):  
Stem Cell ◽  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Cell Clone ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Cell Fraction ◽  
Rt Pcr

Abstract One important question in stem cell biology of childhood acute lymphoblastic leukemia (ALL) is whether immature CD34+CD19− cells are part of the leukemic cell clone. CD34+CD19− cells from the bone marrow of 9 children with TEL/AML1-positive ALL were purified by flow sorting and subjected to reverse transcriptase–polymerase chain reaction (RT-PCR), fluorescence in situ hybridization, and methylcellulose cultures. In 3 of 8 patients analyzed by RT-PCR, noTEL/AML1-positive cells could be detected in the CD34+CD19− cell fraction. Altogether, the percentage of TEL/AML1-positive cells was low: 1.6% (n = 8; SD 2.2%) by nested real-time RT-PCR and 2.5% (n = 5; SD 2.6%) by fluorescence in situ hybridization. This correlated with the percentage of contaminating CD19+ leukemic cells in the CD34+CD19− cell fraction in 6 control sorts (mean 4.6%, SD 3.6%), indicating that the low levels of leukemic cells detected in the CD34+CD19− cell fraction could be attributed to sorter errors. Methylcellulose cultures in 3 patients provided further evidence that CD34+CD19− cells represent a candidate normal cell population. The clonogenicity of the CD34+CD19− cell fraction was similar to normal progenitors, including growth of primitive granulocyte, erythroid, macrophage, megakaryocyte colony-forming units. Each of 92 colonies from cultures with CD34+CD19− cells tested negative for TEL/AML1. In conclusion, our data support the hypothesis that the leukemia inTEL/AML1-positive childhood ALL originates in a CD19+ lymphoid progenitor. This has many therapeutic implications, eg, for purging of autologous stem cell products, flow cytometric monitoring of minimal residual disease, and targeting immunotherapy against the leukemic cell clone.

scholarly journals bcr rearrangement and translocation of the c-abl oncogene in Philadelphia positive acute lymphoblastic leukemia

Blood ◽  
1986 ◽  
Vol 68 (6) ◽  
pp. 1369-1375 ◽  
Author(s):  
A De Klein ◽  
A Hagemeijer ◽  
CR Bartram ◽  
R Houwen ◽  
L Hoefsloot ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Blot Analysis ◽  
Myeloid Leukemia ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cells ◽  
Mrna Transcripts

Abstract The Philadelphia (Ph1) chromosome, the cytogenetic hallmark of chronic myeloid leukemia (CML), has also been detected in a significant number of acute lymphoblastic leukemias (ALL). Using in situ hybridization, we demonstrate that in accordance with observations in CML the Ph1 chromosome in ALL patients is the result of a consistent translocation of the c-abl oncogene to the Ph1 chromosome. Southern blot analysis using bcr probes, however, suggests that Ph1-positive ALL includes heterogeneous leukemic subtypes: six ALL patients showed bcr rearrangements as observed in CML; in three other patients recombination involving 5′ bcr sequences could be demonstrated, but the corresponding translocated 3′ bcr sequences were not detectable. A third group of five patients did not show any bcr rearrangements at all. Northern blot analysis using RNA from three Ph1-positive ALL patients revealed that in the leukemic cells of two patients larger c- abl mRNA transcripts were present, as in CML. In the RNA of one patient without a detectable bcr rearrangement, only the normal c-abl mRNA transcripts are present. The observed heterogeneity in bcr rearrangements of this group of Ph1-positive ALL patients is in contrast with the consistent results obtained in more than 50 Ph1- positive CML patients investigated in chronic and acute states.

scholarly journals bcr rearrangement and translocation of the c-abl oncogene in Philadelphia positive acute lymphoblastic leukemia

Blood ◽  
1986 ◽  
Vol 68 (6) ◽  
pp. 1369-1375 ◽  
Author(s):  
A De Klein ◽  
A Hagemeijer ◽  
CR Bartram ◽  
R Houwen ◽  
L Hoefsloot ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Blot Analysis ◽  
Myeloid Leukemia ◽  
Southern Blot Analysis ◽  
Northern Blot Analysis ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cells ◽  
Mrna Transcripts

The Philadelphia (Ph1) chromosome, the cytogenetic hallmark of chronic myeloid leukemia (CML), has also been detected in a significant number of acute lymphoblastic leukemias (ALL). Using in situ hybridization, we demonstrate that in accordance with observations in CML the Ph1 chromosome in ALL patients is the result of a consistent translocation of the c-abl oncogene to the Ph1 chromosome. Southern blot analysis using bcr probes, however, suggests that Ph1-positive ALL includes heterogeneous leukemic subtypes: six ALL patients showed bcr rearrangements as observed in CML; in three other patients recombination involving 5′ bcr sequences could be demonstrated, but the corresponding translocated 3′ bcr sequences were not detectable. A third group of five patients did not show any bcr rearrangements at all. Northern blot analysis using RNA from three Ph1-positive ALL patients revealed that in the leukemic cells of two patients larger c- abl mRNA transcripts were present, as in CML. In the RNA of one patient without a detectable bcr rearrangement, only the normal c-abl mRNA transcripts are present. The observed heterogeneity in bcr rearrangements of this group of Ph1-positive ALL patients is in contrast with the consistent results obtained in more than 50 Ph1- positive CML patients investigated in chronic and acute states.

scholarly journals Detection of chimeric BCR-ABL genes on bone marrow samples and blood smears in chronic myeloid and acute lymphoblastic leukemia by in situ hybridization

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1922-1928 ◽  
Author(s):  
M Bentz ◽  
G Cabot ◽  
M Moos ◽  
MR Speicher ◽  
A Ganser ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Yeast Artificial Chromosome ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
High Specificity ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Blood Smears

Abstract The presence of BCR-ABL fusion genes has important diagnostic and prognostic implications in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL). The CML-specific chimeric BCR-ABL gene with a break involving the major breakpoint cluster region (M-bcr) of the BCR-gene has been detected by means of fluorescence in situ hybridization (FISH). In this study, we present a FISH protocol that allows the detection of breaks in both the major and the minor breakpoint cluster region (m-bcr). Three hybridization signals of D107F9, a yeast artificial chromosome (YAC)-derived probe spanning the breakpoint regions of the BCR gene, were indicative of the translocation events. To increase the specificity further, this probe was combined with cos-abl 8, a cosmid probe flanking the breakpoint within the ABL gene for dual-color hybridization. Samples of 21 patients with CML, the ALL-derived cell line SUP-B15, and of seven patients with Philadelphia chromosome (Ph1)-positive ALL (three of them with breakpoints within m-bcr) were examined. BCR-ABL fusion was detected in all cases with high specificity (false-positive nuclei: mean, 0.1%). On cytogenetic preparations, the percentages of BCR-ABL- positive interphase cells ranged from 53% to 91%. Comparable efficiencies were achieved on blood smears. In conclusion, hybridization with D107F9 and cos-abl 8 allows unambiguous diagnosis of BCR-ABL genes and is likely to become an important tool for the monitoring of therapies in patients with CML and ALL.

scholarly journals Leukemic cell growth in SCID mice as a predictor of relapse in high- risk B-lineage acute lymphoblastic leukemia

Blood ◽  
1995 ◽  
Vol 85 (4) ◽  
pp. 873-878 ◽  
Author(s):  
FM Uckun ◽  
H Sather ◽  
G Reaman ◽  
J Shuster ◽  
V Land ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
High Risk ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cell ◽  
Scid Mice ◽  
Leukemic Cells ◽  
Newly Diagnosed ◽  
B Lineage ◽  
Human Leukemic Cells ◽  
Overt Leukemia

Mice with severe combined immunodeficiency (SCID) provide a model system to examine the in vivo homing, engraftment, and growth patterns of normal and malignant human hematopoietic cells. The relation between leukemic cell growth in this model and the treatment outcome in patients from whom cells were derived has not been established. Leukemic cells from 42 children with newly diagnosed high-risk B- lineage acute lymphoblastic leukemia were inoculated intravenously into CB.17 SCID mice. Mice were killed at 12 weeks or when they became moribund as a result of disseminated leukemia. All mice were necropsied and subjected to a series of laboratory studies to assess their burden of human leukemic cells. Twenty-three patients whose leukemic cells caused histopathologically detectable leukemia in SCID mice had a significantly higher relapse rate than the 19 patients whose leukemic cells did not (estimated 5-year event-free survival: 29.5% v 94.7%; 95% confidence intervals, 11.2% to 50.7% v 68.1% to 99.2%; P < .0001 by log- rank test). The occurrence of overt leukemia in SCID mice was was a highly significant predictor of patient relapse. The estimated instantaneous risk of relapse for patients whose leukemic cells caused overt leukemia in SCID mice was 21.5-fold greater than that for the remaining patients. Thus, growth of human leukemic cells in SCID mice is a strong and independent predictor of relapse in patients with newly diagnosed high-risk B-lineage acute lymphoblastic leukemia.

scholarly journals A New Subtype of Pre-B Acute Lymphoblastic Leukemia With t(5; 12)(q31q33; p12), Molecularly and Cytogenetically Distinct From t(5; 12) in Chronic Myelomonocytic Leukemia

Blood ◽  
1997 ◽  
Vol 89 (5) ◽  
pp. 1716-1722 ◽  
Author(s):  
Iwona Wlodarska ◽  
Anna Aventı́n ◽  
Júlia Inglés-Esteve ◽  
Daniela Falzetti ◽  
Arnold Criel ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Cell Lines ◽  
Chromosomal Abnormality ◽  
Gene Fusion ◽  
Lymphoblastic Leukemia ◽  
Chronic Myelomonocytic Leukemia ◽  
Myelomonocytic Leukemia

Abstract Translocation t(5; 12)(q33; p13), resulting in an ETV6/PDGFRB gene fusion, is a recurrent chromosomal abnormality associated with chronic myelomonocytic leukemia (CMML). An analogous translocation was also found in four cell lines with features of pre-B acute lymphoblastic leukemia (ALL). Using fluorescence in situ hybridization (FISH) we show here that in three of these cell lines identical complex rearrangements occurred. However, the regions involved on 5q and 12p are different from the breakpoints in CMML, and the translocation is accompanied by seemingly identical cryptic deletions of both 5q and 12p chromosome sequences in all analyzed pre-B ALL cell lines. The similar cytogenetic, FISH, and immunophenotyping findings in the three cell lines suggest that the t(5; 12)(q31q33; p12) defines a new entity of pre-B ALL.

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