EuroFlow Lymphoid Screening Tube (LST) data base for automated identification of blood lymphocyte subsets

Autor: Alejando Hernández, Noemí Muñoz, Bruno Paiva, Margarida Lima, Rafael Fluxa, Lukasz Sedek, Ana Helena Santos, Alberto Orfao, Quentin Lecrevisse, Juan Ramón Hernández Hernández, Georgiana Grigore, Paula Fernandez, Vincent H.J. van der Velden, Sebastian Böttcher, Susana Barrena, Juan Flores-Montero, Julia Almeida, Jacques J.M. van Dongen
Přispěvatelé: Immunology, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad (España), European Commission
Rok vydání: 2019
Předmět:
Zdroj: Journal of Immunological Methods, 475:Unsp 112662. Elsevier
Digital.CSIC. Repositorio Institucional del CSIC
instname
Dadun. Depósito Académico Digital de la Universidad de Navarra
Journal of Immunological Methods, 475. ELSEVIER
ISSN: 0022-1759
Popis: © 2019 The Authors.
In recent years the volume and complexity of flow cytometry data has increased substantially. This has led to a greater number of identifiable cell populations in a single measurement. Consequently, new gating strategies and new approaches for cell population definition are required. Here we describe how the EuroFlow Lymphoid Screening Tube (LST) reference data base for peripheral blood (PB) samples was designed, constructed and validated for automated gating of the distinct lymphoid (and myeloid) subsets in PB of patients with chronic lymphoproliferative disorders (CLPD). A total of 46 healthy/reactive PB samples which fulfilled pre-defined technical requirements, were used to construct the LST-PB reference data base. In addition, another set of 92 PB samples (corresponding to 10 healthy subjects, 51 B-cell CLPD and 31 T/NK-cell CLPD patients), were used to validate the automated gating and cell-population labeling tools with the Infinicyt software. An overall high performance of the LST-PB data base was observed with a median percentage of alarmed cellular events of 0.8% in 10 healthy donor samples and of 44.4% in CLPD data files containing 49.8% (range: 1.3–96%) tumor cells. The higher percent of alarmed cellular events in every CLPD sample was due to aberrant phenotypes (75.6% cases) and/or to abnormally increased cell counts (86.6% samples). All 18 (22%) data files that only displayed numerical alterations, corresponded to T/NK-cell CLPD cases which showed a lower incidence of aberrant phenotypes (41%) vs B-cell CLPD cases (100%). Comparison between automated vs expert-bases manual classification of normal (r2 = 0.96) and tumor cell populations (rho = 0.99) showed a high degree of correlation. In summary, our results show that automated gating of cell populations based on the EuroFlow LST-PB data base provides an innovative, reliable and reproducible tool for fast and simplified identification of normal vs pathological B and T/NK lymphocytes in PB of CLPD patients.
This work was supported by the CB16/12/00400 and CB16/12/00369 grants, CIBER-ONC, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain and FONDOS FEDER; DTS 15/00119 grant, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain and FONDOS FEDER; and, RETOS (RTC–2016–4865–1) grant, Sociedad de la Acción Estratégica de Economía y Sociedad Digital-Impulso Tecnológico; Ministerio de Economía y Competitividad, Madrid, Spain.
Databáze: OpenAIRE
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