Flow cytometric identification of candidate normal stem cell populations in CD45‐negative B‐cell precursor acute lymphoblastic leukaemia

Vormoor, Josef; Baersch, Gudrun; Baumann, M; Ritter, J.; Jürgens, Heribert

doi:10.1046/j.1365-2141.1998.00610.x

Cited by 13 publications

(23 citation statements)

References 38 publications

(41 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Flow cytometric investigations showed that primitive progenitor/stem cells have a phenotype that is very distinct from the leukemic clone (10,11). These observations together with molecular analyses in good-prognosis ALL/t(12/21) demonstrating that primitive CD34 + CD19 À do not carry the leukemia-specific chromosomal translocation (12) supported this hypothesis.…”

Section: Introductionmentioning

confidence: 62%

“…Its expression is an early event in B-cell development (30) directly following DJ H but preceding or simultaneously occurring with VDJ H rearrangement (31). Analysis of immature CD34 + CD19 À cells for expression of CD45, CD117, and CD133 in the bone marrow from children with B-cell precursor ALL showed that these cells have an immunophenotype similar to normal progenitor/stem cells (10,11). Accordingly, these CD34 + CD19 À cells have a similar clonogenicity in myeloid colony assays compared with CD34 + CD19 À cells from normal bone marrow (12).…”

Section: Resultsmentioning

confidence: 95%

“…Through extensive flow cytometric studies of immature cell populations in childhood ALL, we have shown that expression of the B-cell antigen CD19 is a suitable marker to distinguish immature progenitor/stem cells from the bulk leukemic population (10,11). CD19 is part of the CD19/CD21 complex that modulates B-cell receptor signaling (29).…”

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Leukemic Stem Cells in Childhood High-Risk ALL/t(9;22) and t(4;11) Are Present in Primitive Lymphoid-Restricted CD34+CD19− Cells

et al. 2005

View full text Add to dashboard Cite

Open questions in the pathogenesis of childhood acute lymphoblastic leukemia (ALL) are which hematopoietic cell is target of the malignant transformation and whether primitive stem cells contribute to the leukemic clone. Although good-prognosis ALL is thought to originate in a lymphoid progenitor, it is unclear if this applies to high-risk ALL. Therefore, immature CD34 + CD19 À bone marrow cells from 8 children with ALL/t(9;22) and 12 with ALL/t(4;11) were purified and analyzed by fluorescence in situ hybridization, reverse transcription-PCR (RT-PCR), and colony assays. Fiftysix percent (n = 8, SD 31%) and 68% (n = 12, SD 26%) of CD34 + CD19 À cells in ALL/t(9;22) and ALL/t(4;11), respectively, carried the translocation. In addition, 5 of 168 (3%) and 22 of 228 (10%) myeloerythroid colonies expressed BCR/ABL and MLL/AF4. RT-PCR results were confirmed by sequence analysis. Interestingly, in some patients with ALL/t(4;11), alternative splicing was seen in myeloid progenitors compared with the bulk leukemic population, suggesting that these myeloid colonies might be part of the leukemic cell clone. Fluorescence in situ hybridization analysis, however, shows that none of these myeloid colonies (0 of 41 RT-PCRpositive colonies) originated from a progenitor cell that carries the leukemia-specific translocation. Thus, leukemic, translocation-positive CD34 + CD19 À progenitor/stem cells that were copurified by cell sorting were able to survive in these colony assays for up to 28 days allowing amplification of the respective fusion transcripts by sensitive RT-PCR. In conclusion, we show that childhood high-risk ALL/t(9;22) and t(4;11) originate in a primitive CD34 + CD19 À progenitor/stem cell without a myeloerythroid developmental potential. (Cancer Res 2005; 65(4): 1442-9)

show abstract

Section: Introductionmentioning

confidence: 62%

Section: Resultsmentioning

confidence: 95%

See 1 more Smart Citation

Leukemic Stem Cells in Childhood High-Risk ALL/t(9;22) and t(4;11) Are Present in Primitive Lymphoid-Restricted CD34+CD19− Cells

et al. 2005

View full text Add to dashboard Cite

show abstract

“…A possible biological explanation could be the presence of low numbers of BCR-ABL-positive leukemic progenitors with low or absent expression of the CD19 antigen 40 thus escaping the purging procedure.…”

Section: Discussionmentioning

confidence: 99%

Purging in BCR-ABL-positive acute lymphoblastic leukemia using immunomagnetic beads: comparison of residual leukemia and purging efficiency in bone marrow vs peripheral blood stem cells by semiquantitative polymerase chain reaction

Atta

Fauth

et al. 2000

Bone Marrow Transplant

View full text Add to dashboard Cite

Summary:Twenty autologous bone marrow (BM) and 25 peripheral blood stem cell (PBSC) grafts were collected from a total of 40 consecutive patients with BCR-ABL + acute lymphoblastic leukemia (ALL) in first (n = 37) or second (n = 3) complete morphological remission and subsequently purged with a cocktail of anti-CD19, -CD10, AB4 MoAbs and immunomagnetic beads (IMB). Residual BCR-ABL-positive cells before purging were detected in 19 of 20 BM grafts at a median of 4 (range 0-6) logs and in 17 of 25 evaluable PBSC grafts at a median of 1 (range 0-3) log above the limit of detection assessed by a semiquantitative limiting log 10 -dilution RT-PCR (P Ͻ 0.0001). IMB purging depleted a median of 2.5 (range 1-4) log of residual BCR-ABL + cells from BM and a median of 1 (range 0-2) log from PBSC grafts, achieving RT-PCR negativity in 1/20 BM and 12/25 PBSC grafts after purging. Cell recoveries were 62% and 86% (P Ͻ 0.0001) of MNC and 74% and 97% (P = 0.065) of CD34 + cells after BM and PBSC purging, respectively. BM purging was superior using the triple MoAb cocktail which depleted 2.64 ؎ 0.4 log (n = 14) compared to 1.6 ؎ 0.4 log (n = 5) using the MoAb cocktail not including AB4 (P = 0.02). We conclude that unpurged BM grafts contain 2-3 log more residual BCR-ABL + cells than unpurged PBSC grafts and that purging efficacy is superior in BM compared to PBSC grafts, but median titers in purged BM grafts still exceed those in purged PBSC grafts. Bone Marrow Transplantation (2000) 25, 97-104.

show abstract

“…To quantify and characterize human Ewing tumor cells infiltrating the murine marrow, two-and three-color flow cytometry was performed according to standard procedures (34,35). In short, bone marrow cells were incubated with saturating amounts of FITC-, PE-, and CyChrome-conjugated antibodies for 20 min at room temperature.…”

Section: Ewing Tumor-nod/scid Mouse Modelmentioning

confidence: 99%

Establishment of an In Vivo Model for Pediatric Ewing Tumors by Transplantation into NOD/ scid Mice

et al. 2001

View full text Add to dashboard Cite

Ewing tumors are a clinically heterogeneous group of childhood sarcomas that represent a paradigm for understanding solid tumor biology, as they are the first group of sarcomas for which a chromosome translocation has been characterized at the molecular level. However, the biologic organization of the tumor, especially the processes that govern proliferation, differentiation, and metastasis of primitive tumor stem cells is poorly understood. Therefore, to develop a biologically relevant in vivo model, five different Ewing tumor cell lines and primary tumor cells from three patients were transplanted into immune-deficient mice via intravenous injection. NOD/scid mice that carry a complex immune deficiency and thus nearly completely lack the ability to reject human cells were used as recipients. Overall, 26 of 52 mice (50%) transplanted with VH-64, WE-68, CADO-ES1, TC-71, and RM-82 cells and 4 of 10 mice (40%) transplanted with primary tumor cells engrafted. Moreover, primary cells that did not grow in vitro proliferated in mice. The pattern of metastasis was similar to that in patients with frequent metastases in lungs (62%), bone marrow (30%), and bone (23%). Using limiting dilution experiments, the frequency of the engraftment unit was estimated at 1 Ewing tumor-initiating cell in 3 ϫ 10 5 VH-64 cells. These data demonstrate that we have been able to establish an in vivo model that recapitulates many aspects of growth and progression of human Ewing tumors. For the first time, this model provides the opportunity to identify and characterize primitive in vivo clonogenic solid tumor stem cells. This model will, therefore, be instrumental in studying many aspects of tumor cell biology, including organ-selective metastasis and tumor angiogenesis. The family of Ewing tumors is a clinically heterogeneous group of tumors including Ewing's sarcoma of the bone, Askin tumors of the thoracic wall, and PNET affecting children, adolescents, and young adults. Despite this clinical heterogeneity, Ewing tumors represent a paradigm for understanding the biology of human solid tumors, as they are the first group of sarcomas for which a chromosome translocation has been characterized at the molecular level (1, 2). This translocation, t(11;22), leads to the fusion of the EWS gene at 22q12, encoding for a putative RNA binding protein, to FLI1 at 11q24, an ETS-related gene coding for a transcription factor. Different breakpoints occur with in-frame fusion of EWS exons 7-10 to FLI1 exons 4 -9 (3). The two most common combinations are EWS exon 7/FLI1 exon 6 (termed type 1 fusion) and EWS exon 7/FLI1 exon 5 (type 2 fusion). The biologic relevance of the different breakpoints is indicated by two retrospective analyses demonstrating that patients with nonmetastatic disease and a type 1 fusion had a better clinical outcome than patients with other exon combinations (4, 5). In another 5%-10% of Ewing tumor patients, EWS is fused to others members of the ETS family, most frequently ERG at 21q22 (6) or more rarely ETV1, E1AF and FEV (7-10). O...

show abstract

Flow cytometric identification of candidate normal stem cell populations in CD45‐negative B‐cell precursor acute lymphoblastic leukaemia

Cited by 13 publications

References 38 publications

Leukemic Stem Cells in Childhood High-Risk ALL/t(9;22) and t(4;11) Are Present in Primitive Lymphoid-Restricted CD34+CD19− Cells

Leukemic Stem Cells in Childhood High-Risk ALL/t(9;22) and t(4;11) Are Present in Primitive Lymphoid-Restricted CD34+CD19− Cells

Purging in BCR-ABL-positive acute lymphoblastic leukemia using immunomagnetic beads: comparison of residual leukemia and purging efficiency in bone marrow vs peripheral blood stem cells by semiquantitative polymerase chain reaction

Establishment of an In Vivo Model for Pediatric Ewing Tumors by Transplantation into NOD/ scid Mice

Contact Info

Product

Resources

About