Precise classification of acute leukemia (AL) is crucial for adequate treatment. EuroFlow has previously designed an AL orientation tube (ALOT) to guide towards the relevant classification panel (T-cell acute lymphoblastic leukemia (T-ALL), B-cell precursor (BCP)-ALL and/or acute myeloid leukemia (AML)) and final diagnosis. Now we built a reference database with 656 typical AL samples (145 T-ALL, 377 BCP-ALL, 134 AML), processed and analyzed via standardized protocols. Using principal component analysis (PCA)-based plots and automated classification algorithms for direct comparison of single-cells from individual patients against the database, another 783 cases were subsequently evaluated. Depending on the database-guided results, patients were categorized as: (i) typical T, B or Myeloid without or; (ii) with a transitional component to another lineage; (iii) atypical; or (iv) mixed-lineage. Using this automated algorithm, in 781/783 cases (99.7%) the right panel was selected, and data comparable to the final WHO-diagnosis was already provided in >93% of cases (85% T-ALL, 97% BCP-ALL, 95% AML and 87% mixed-phenotype AL patients), even without data on the full-characterization panels. Our results show that database-guided analysis facilitates standardized interpretation of ALOT results and allows accurate selection of the relevant classification panels, hence providing a solid basis for designing future WHO AL classifications.

Biological Hematology AP HP Necker Enfants Malades Paris France

Cancer Research Center Salamanca Spain and Centro de Investigación Biomédica en Red de Cáncer Instituto Carlos 3 Madrid Spain

Clinica Universidad de Navarra Centro De Investigaciones Medicas Aplicadas University of Navarra Pamplona Spain

CLIP Childhood Leukaemia Investigation Prague Department of Paediatric Haematology and Oncology 2nd Faculty of Medicine Charles University University Hospital Motol Prague Czech Republic

COPPE Computation Engineering and Systems Post graduation Federal University of Rio de Janeiro Rio de Janeiro Brazil

Cytognos SL Salamanca Spain

Department of Hematology University of Schleswig Holstein Campus Kiel Kiel Germany

Department of Immunohematology and Blood Transfusion Leiden University Medical Center Leiden The Netherlands

Department of Immunology Erasmus MC University Medical Center Rotterdam Rotterdam The Netherlands

Department of Microbiology and Immunology Zabrze Medical University of Silesia Katowice Poland

Department of Pediatric Hematology and Oncology Zabrze Medical University of Silesia Katowice Poland

Department of Pediatrics Faculty of Medicine Federal University of Rio de Janeiro Rio de Janeiro Brazil

Dutch Childhood Oncology Group The Hague The Netherlands

Hematology Service University Hospital of Salamanca Salamanca Spain

Institute for Laboratory Medicin Kantonsspital Aarau AG Aarau Switzerland

Tettamanti Research Center Pediatric Clinic University of Milano Bicocca Monza Italy

Université Paris Descartes Sorbonne Paris Cité Institut Necker Enfants Malades INSERM UMR1151 Paris France

Zobrazit více v PubMed

Borowitz MJ, JKC Chan B lymphoblastic leukaemia/lymphoma. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J, Vardiman JW (eds). WHO classification of Tumours of Haematopoietic and Lymphoid Tissues, 4th edn. International Agency for Research on Cancer: Lyon, 2008, pp 168–175.

Borowitz MJ, JKC Chan T lymphoblastic leukaemia/lymphoma. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J, Vardiman JW (eds). WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues, 4th edn. International Agency for Research on Cancer: Lyon, 2008, pp 176–178.

Borowitz MJ, Bene MC, Harris NL, Porwitt A, Matutes E. Acute leukaemias of ambiguous lineage. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J, Vardiman JW (eds). WHO Classification of Tumours ofHaematopoietic and Lymphoid Tissues, 4th edn. International Agency for Research on Cancer: Lyon, 2008, pp 150–155.

Mejstrikova E, Volejnikova J, Fronkova E, Zdrahalova K, Kalina T, Sterba J et al. Prognosis of children with mixed phenotype acute leukemia treated on the basis of consistent immunophenotypic criteria. Haematologica 2010; 95: 928–935. PubMed PMC

van den Ancker W, Terwijn M, Westers TM, Merle PA, van Beckhoven E, Drager AM et al. Acute leukemias of ambiguous lineage: diagnostic consequences of the WHO2008 classification. Leukemia 2010; 24: 1392–1396. PubMed

Flores-Montero J, Sanoja-Flores L, Paiva B, Puig N, Garcia-Sanchez O, Bottcher S et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia 2017; 31: 2094–2103. PubMed PMC

Kalina T, Flores-Montero J, van der Velden VH, Martin-Ayuso M, Bottcher S, Ritgen M et al. EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols. Leukemia 2012; 26: 1986–2010. PubMed PMC

van Dongen JJ, Lhermitte L, Bottcher S, Almeida J, van der Velden VH, Flores-Montero J et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia 2012; 26: 1908–1975. PubMed PMC

Flores-Montero J, Flores LS, Paiva B, Puig N, Garcia-Sanchez O, Bottcher S et al. Next generation flow (NGF) for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia 2017; 31: 2094–2103. PubMed PMC

Theunissen P, Mejstrikova E, Sedek L, van der Sluijs-Gelling AJ, Gaipa G, Bartels M et al. Standardized flow cytometry for highly sensitive MRD measurements in B-cell acute lymphoblastic leukemia. Blood 2017; 129: 347–357. PubMed PMC

Han K, Kim Y, Lee J, Lim J, Lee KY, Kang CS et al. Human basophils express CD22 without expression of CD19. Cytometry 1999; 37: 178–183. PubMed

Kalina T, Flores-Montero J, Lecrevisse Q, Pedreira CE, van der Velden VH, Novakova M et al. Quality assessment program for EuroFlow protocols: summary results of four-year (2010-2013) quality assurance rounds. Cytometry A 2015; 87: 145–156. PubMed

Costa ES, Pedreira CE, Barrena S, Lecrevisse Q, Flores J, Quijano S et al. Automated pattern-guided principal component analysis vs expert-based immunophenotypic classification of B-cell chronic lymphoproliferative disorders: a step forward in the standardization of clinical immunophenotyping. Leukemia 2010; 24: 1927–1933. PubMed PMC

Steiner M, Attarbaschi A, Dworzak M, Strobl H, Pickl W, Kornmuller R et al. Cytochemically myeloperoxidase positive childhood acute leukemia with lymphoblastic morphology treated as lymphoblastic leukemia. J Pediatr Hematol Oncol 2010; 32: e4–e7. PubMed

Pedreira CE, Costa ES, Lecrevisse Q, van Dongen JJ, Orfao A, EuroFlow C. Overview of clinical flow cytometry data analysis: recent advances and future challenges. Trends Biotechnol 2013; 31: 415–425. PubMed

Slamova L, Starkova J, Fronkova E, Zaliova M, Reznickova L, van Delft FW et al. CD2-positive B-cell precursor acute lymphoblastic leukemia with an early switch to the monocytic lineage. Leukemia 2014; 28: 609–620. PubMed

Automated identification of leukocyte subsets improves standardization of database-guided expert-supervised diagnostic orientation in acute leukemia: a EuroFlow study

EuroFlow-Based Flowcytometric Diagnostic Screening and Classification of Primary Immunodeficiencies of the Lymphoid System