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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

AuthorsTheunissen, Prisca; Sedek, Lukasz; De Haas, Valerie; Szczepanski, Tomasz; Sluijs-Gelling, Alita J. van der; Mejstrikova, Ester; Novákova, Michaela; Kalina, Tomas; Lécrevisse, Quentin; Orfao, Alberto ; Lankester, Arjan C.; Dongen, J. J. M. van; Velden, Vincent H. J. van der
KeywordsB cells
Acute leukaemia
Flow cytometry
Minimal residual disease
Issue Date2017
PublisherJohn Wiley & Sons
CitationBritish Journal of Haematology 178(2): 257-266 (2017)
AbstractFlow 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 pre-B-I cell immunophenotype. These CD34 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 pre-B-I cells. Our results indicate that newly identified CD34 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 pre-B-I cells are relatively abundant, should be used as reference frame in flow cytometric MRD measurements.
Identifiersdoi: 10.1111/bjh.14682
e-issn: 1365-2141
issn: 0007-1048
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