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Detailed immunophenotyping of B-cell precursors in regenerating bone marrow of acute lymphoblastic leukaemia patients: implications for minimal residual disease detection

PMJ. Theunissen, L. Sedek, V. De Haas, T. Szczepanski, A. Van Der Sluijs, E. Mejstrikova, M. Nováková, T. Kalina, Q. Lecrevisse, A. Orfao, AC. Lankester, JJM. van Dongen, VHJ. Van Der Velden, . ,

. 2017 ; 178 (2) : 257-266. [pub] 20170417

Language English Country England, Great Britain

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc17030879

Grant support
NV15-28525A MZ0 CEP Register

Flow cytometric detection of minimal residual disease (MRD) in children with B-cell precursor acute lymphoblastic leukaemia (BCP-ALL) requires immunophenotypic discrimination between residual leukaemic cells and B-cell precursors (BCPs) which regenerate during therapy intervals. In this study, EuroFlow-based 8-colour flow cytometry and innovative analysis tools were used to first characterize the immunophenotypic maturation of normal BCPs in bone marrow (BM) from healthy children, resulting in a continuous multiparametric pathway including transition stages. This pathway was subsequently used as a reference to characterize the immunophenotypic maturation of regenerating BCPs in BM from children treated for BCP-ALL. We identified pre-B-I cells that expressed low or dim CD34 levels, in contrast to the classical CD34(high) pre-B-I cell immunophenotype. These CD34(-dim) pre-B-I cells were relatively abundant in regenerating BM (11-85% within pre-B-I subset), while hardly present in healthy control BM (9-13% within pre-B-I subset; P = 0·0037). Furthermore, we showed that some of the BCP-ALL diagnosis immunophenotypes (23%) overlapped with CD34(-dim) pre-B-I cells. Our results indicate that newly identified CD34(-dim) pre-B-I cells can be mistaken for residual BCP-ALL cells, potentially resulting in false-positive MRD outcomes. Therefore, regenerating BM, in which CD34(-dim) pre-B-I cells are relatively abundant, should be used as reference frame in flow cytometric MRD measurements.

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$a Flow cytometric detection of minimal residual disease (MRD) in children with B-cell precursor acute lymphoblastic leukaemia (BCP-ALL) requires immunophenotypic discrimination between residual leukaemic cells and B-cell precursors (BCPs) which regenerate during therapy intervals. In this study, EuroFlow-based 8-colour flow cytometry and innovative analysis tools were used to first characterize the immunophenotypic maturation of normal BCPs in bone marrow (BM) from healthy children, resulting in a continuous multiparametric pathway including transition stages. This pathway was subsequently used as a reference to characterize the immunophenotypic maturation of regenerating BCPs in BM from children treated for BCP-ALL. We identified pre-B-I cells that expressed low or dim CD34 levels, in contrast to the classical CD34(high) pre-B-I cell immunophenotype. These CD34(-dim) pre-B-I cells were relatively abundant in regenerating BM (11-85% within pre-B-I subset), while hardly present in healthy control BM (9-13% within pre-B-I subset; P = 0·0037). Furthermore, we showed that some of the BCP-ALL diagnosis immunophenotypes (23%) overlapped with CD34(-dim) pre-B-I cells. Our results indicate that newly identified CD34(-dim) pre-B-I cells can be mistaken for residual BCP-ALL cells, potentially resulting in false-positive MRD outcomes. Therefore, regenerating BM, in which CD34(-dim) pre-B-I cells are relatively abundant, should be used as reference frame in flow cytometric MRD measurements.
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$a Sedek, Lukasz $u Department of Paediatric Haematology and Oncology, Zabrze, Poland. Medical University of Silesia (SUM), Katowice, Poland.
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$a De Haas, Valerie $u Dutch Childhood Oncology Group, The Hague, the Netherlands.
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$a Szczepanski, Tomasz $u Department of Paediatric Haematology and Oncology, Zabrze, Poland. Medical University of Silesia (SUM), Katowice, Poland.
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$a Van Der Sluijs, Alita $u Dutch Childhood Oncology Group, The Hague, the Netherlands.
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$a Mejstrikova, Ester $u Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) and University Hospital Motol, Prague, Czech Republic.
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$a Nováková, Michaela $u Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) and University Hospital Motol, Prague, Czech Republic.
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$a Kalina, Tomas $u Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) and University Hospital Motol, Prague, Czech Republic.
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$a Lecrevisse, Quentin $u Cancer Research Centre (IBMCC-CSIC), Department of Medicine and Cytometry Service, University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain.
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$a Lankester, Arjan C $u Department of Paediatrics, Leiden University Medical Centre, Leiden, the Netherlands.
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$a van Dongen, Jacques J M $u Department of Immunology, Erasmus MC, University Medical Centre Rotterdam, Rotterdam, the Netherlands. Department of Immunohaematology and Blood Transfusion, Leiden University Medical Centre, Leiden, the Netherlands.
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