Minimal residual disease in acute leukaemia

Gerhartz, H.; Schmetzer, Helga

doi:10.1016/0277-5379(91)90199-n

Cited by 6 publications

(12 citation statements)

References 4 publications

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“…LAK and BM cells were mixed in a proportion of 5:1. After 4 h, the cell mixtures were mixed with agar, plated on special slides (BioRad) and stimulated by GM-CSF (500 U/ml) for a 7-day colony assay [4]. In this colony assay LAK cells formed colonies under the influence of GM-CSF.…”

Section: Culture Of Lak Cells and Coculture With Autologous Bm Cellsmentioning

confidence: 99%

“…After 7 days of culture, colony counts were calculated. Moreover, immunologically stained colonies were counted after in situ incubation with a panel of different antibodies [4]. The total cell and colony counts before and after 14 days of cytokine incubation (Cytok 1, Cytok 2 or ISC/FCS as a control) or LAK cell treatment were evaluated to estimate the reductive effect on leukemic colonies as well as the proliferation-stimulating effect of those cytokine cocktails and LAK cells.…”

Section: Immunophenotyping Of Agar Colonies In Situmentioning

confidence: 99%

“…To study the viability of BM-MNC as well as the influence of cytokines and autologous LAK cells on leukemic colonies, we cultured mononuclear cells for 7 days in a miniaturized culture assay directly on sterilized and specially prepared slides obtained from BioRad as previously described [4]. 3 !…”

Section: Immunophenotyping Of Agar Colonies In Situmentioning

confidence: 99%

“…Flow cytometric analyses were performed on uncultured and cultured BM-MNC in order to estimate the percentage of blast cells (positive for CD34), granulocytes (positive for CD15 and CDw65), T cells (positive for CD3) and B cells (positive for CD19 and CD20) [3,4]. Antibodies conjugated with fluorescent dyes -phycoerythrin (PE) or fluorescein isothiocyanate (FITC) -were used: CD34 (class III clone 581, PE-labeled; Immunotech), CD15 (FITC or PE-labeled; Sigma), CDw65 (FITC-labeled; Ortho), CD19 (PE-labeled; Ortho), CD20 (PE-labeled, Dako) or CD3 (FITC-labeled; Ortho).…”

Section: Surface Marker Analysesmentioning

confidence: 99%

“…Leukemic cell populations can be identified by flow cytometry using a panel of different antibodies [3]. At colony level, leukemic colonies can be distinguished from nonleukemic colonies by culture of bone marrow mononuclear cells (BM-MNC) in a clonogenic assay with consecutive immunophenotyping in situ [4]. About 70% of patients with AML in complete remission relapse within 2 years.…”

Section: Introductionmentioning

confidence: 99%

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Influence of Cytokines and Autologous Lymphokine-Activated Killer Cells on Leukemic Bone Marrow Cells and Colonies in AML

Braun

Gerhartz²,

Schmetzer³

2001

Acta Haematol

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We have already shown that cytokine cocktails (IL-1ß, IL-3, IL-6, SCF, GM-CSF) and/or lymphokine-activated killer (LAK) cells can reduce the amounts of clonal, CD34-positive mononuclear bone marrow cells (BM-MNC) in acute myeloid leukemia (AML). In addition, the influence of those cocktails and/or LAK cells on the clonogenic potential of AML BM-MNC was investigated. BM colonies cultured in agar during different stages of the disease were immunophenotyped in situ: 17 patients at diagnosis, 14 patients in complete remission, 8 patients at relapse, 8 healthy donors. A significant reduction in leukemic cells and colonies positive for CD34 after in vitro culture of BM-MNC with cytokine cocktails was achieved with all samples obtained at diagnosis (n = 8, p ! 0.01), in 6 of 8 cases in complete remission but only in 2 of 6 cases at relapse. Cytokine cocktails stimulated granulopoiesis as well as B and T lymphopoiesis. Colonies with leukemic phenotype could never be detected in healthy BM. A significant reduction in leukemic colonies was achieved by coculture of BM-MNC (uncultured or cytokine precultured) with autologous LAK cells in all 4 cases at diagnosis and in 1 case at relapse. An additive effect of in vitro cytokine preincubation of BM samples on the leukemiareducing effect of LAK cells could be demonstrated in all samples studied (p ! 0.001; diagnosis: n = 10, relapse: n = 3, complete remission: n = 7). Patients had a better prognosis if CD34-positive colonies in AML could be reduced by cytokine incubation (p = 0.03) or coculture with autologous LAK cells in vitro (p = 0.04). Our data show that cytokines as well as LAK cells alone and in combination can reduce, however not eliminate clonogenic AML cells. Such mechanisms might be responsible for maintaining stable remissions in AML.

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Section: Culture Of Lak Cells and Coculture With Autologous Bm Cellsmentioning

confidence: 99%

Section: Immunophenotyping Of Agar Colonies In Situmentioning

confidence: 99%

Section: Immunophenotyping Of Agar Colonies In Situmentioning

confidence: 99%

Section: Surface Marker Analysesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Influence of Cytokines and Autologous Lymphokine-Activated Killer Cells on Leukemic Bone Marrow Cells and Colonies in AML

Braun

Gerhartz²,

Schmetzer³

2001

Acta Haematol

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Detection of Minimal Disease in Hematological Malignancies

Wilmanns¹,

Gerhartz²,

Schmetzer³

et al. 1992

Cancer Diagnosis

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GM-CSF stimulates proliferation of clonal leukemic bone marrow cells in acute myeloid leukemia (AML) in vitro

Schmetzer¹,

Gerhartz

Wilmanns³

1999

Annals of Hematology

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Granulocyte-macrophage colony-stimulating factor (GM-CSF) is known to stimulate granulocytes, monocytes, and macrophages. We studied the effect of GM-CSF on (clonal) bone marrow (BM) cells obtained from AML patients after 7 days of culture in vitro: BM samples were obtained from 19 AML patients at diagnosis (DIA), from two patients with persisting disease (PERS), from eight patients in complete remission (CR), and from 12 healthy donors. Flow-cytometric comparison of differentiated, CD 15-positive cells or of CD34-positive blast cells before and after cultivation showed that the proportion of CD15-positive cells was increased in nine of 12 healthy BM samples, in 14 of 19 cases at DIA, in one of three cases during PERS, and in five of six cases in CR of AML. The proportion of CD34-positive cells was increased in one of 12 healthy BM samples, in seven of 19 cases at DIA, in one of two cases during PERS, and in three of seven cases in CR of AML. Southern blot analysis (SBA) performed in six cases during the course of AML, before and after cell culture, showed that clonal DNA increased after GM-CSF treatment in three of five cases studied at DIA, in six of nine cases studied in CR, in the one case studied at PERS, and in the one studied at relapse (REL). In one case of trisomy 8 at DIA a normal karyotype was demonstrated in CR. However, after 7 days of cultivation of the cells in GM-CSF the trisomy 8 was detected in two of 17 metaphases isolated from colony-cells from methylcellulose cultures. Our data show that a 7-day treatment of BM cells with GM-CSF induced a differentiation of healthy and leukemic BM cells in the great majority of cases. An enrichment of CD34-positive cells was not achieved in healthy BM samples. However, in 70% of the cases in CR and in 30% of the cases at DIA of AML, clonal CD34-positive cells were enriched. This means that GM-CSF stimulates ('primes') leukemic cell growth in vitro.

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Minimal residual disease in acute leukaemia

Cited by 6 publications

References 4 publications

Influence of Cytokines and Autologous Lymphokine-Activated Killer Cells on Leukemic Bone Marrow Cells and Colonies in AML

Influence of Cytokines and Autologous Lymphokine-Activated Killer Cells on Leukemic Bone Marrow Cells and Colonies in AML

Detection of Minimal Disease in Hematological Malignancies

GM-CSF stimulates proliferation of clonal leukemic bone marrow cells in acute myeloid leukemia (AML) in vitro

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