title: Minimal residual disease analysis by eight-color flow cytometry in relapsed childhood acute lymphoblastic leukemia.
abstract: Multiparametric flow cytometry is an alternative approach to the polymerase chain reaction method for evaluating minimal residual disease in treatment protocols for primary acute lymphoblastic leukemia. Given considerable differences between primary and relapsed acute lymphoblastic leukemia treatment regimens, flow cytometric assessment of minimal residual disease in relapsed leukemia requires an independent comprehensive investigation. In the present study we addressed evaluation of minimal residual disease by flow cytometry in the clinical trial for childhood relapsed acute lymphoblastic leukemia using eight-color flow cytometry. The major challenge of the study was to reliably identify low amounts of residual leukemic cells against the complex background of regeneration, characteristic of follow-up samples during relapse treatment. In a prospective study of 263 follow-up bone marrow samples from 122 patients with B-cell precursor acute lymphoblastic leukemia, we tested various B-cell markers, adapted the antibody panel to the treatment protocol, and evaluated its performance by a blinded parallel comparison with the polymerase chain reaction data. The resulting eight-color single-tube panel showed a consistently high overall concordance (P<0.001) and, under optimal conditions, sensitivity similar to that of the reference polymerase chain reaction method. Overall, evaluation of minimal residual disease by flow cytometry can be successfully integrated into the clinical management of relapsed childhood acute lymphoblastic leukemia either as complementary to the polymerase chain reaction or as an independent risk stratification tool. ALL-REZ BFM 2002 clinical trial information: NCT00114348.
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany leonid.karawajew@charite.de.
authors.forename: Leonid
authors.initials: L
authors.lastname: Karawajew
authors.affiliation: St. Anna Children's Hospital and Children's Cancer Research Institute, Department of Pediatrics, Medical University of Vienna, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Dworzak
authors.affiliation: Robert-Roessle-Clinic in the HELIOS Klinikum Berlin, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Ratei
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Rhein
authors.affiliation: Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, Ospedale San Gerardo, Monza, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gaipa
authors.affiliation: Laboratory of Pediatric Onco-Hematology, Department of Pediatrics, University Hospital of Padova, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Buldini
authors.affiliation: Laboratory of Pediatric Onco-Hematology, Department of Pediatrics, University Hospital of Padova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Basso
authors.affiliation: Department of Pediatric Hematology and Oncology, Charles University 2 Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: Ondrej
authors.initials: O
authors.lastname: Hrusak
authors.affiliation: Robert-Roessle-Clinic in the HELIOS Klinikum Berlin, Germany.
authors.forename: Wolf-Dieter
authors.initials: WD
authors.lastname: Ludwig
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Henze
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Seeger
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany.
authors.forename: Arend
authors.initials: A
authors.lastname: von Stackelberg
authors.affiliation: Department of Pediatric Hematology and Oncology, Charles University 2 Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
authors.forename: Ester
authors.initials: E
authors.lastname: Mejstrikova
authors.affiliation: Department of Pediatric Oncology/Hematology, Charité Universitätsmedizin, Berlin, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Eckert
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D000970:Antineoplastic Agents / Q000627:therapeutic use; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000276:immunology; D001853:Bone Marrow / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D002675:Child, Preschool; D004253:DNA Nucleotidylexotransferase / Q000235:genetics / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping; D007223:Infant; D018365:Neoplasm, Residual; D016133:Polymerase Chain Reaction; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000276:immunology; D012008:Recurrence
chemical_list: D015703:Antigens, CD; D000970:Antineoplastic Agents; C000595707:BCL2 protein, human; D014408:Biomarkers, Tumor; D019253:Proto-Oncogene Proteins c-bcl-2; D004253:DNA Nucleotidylexotransferase