EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes
Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and mult...
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| Published in | Leukemia Vol. 26; no. 9; pp. 1908 - 1975 |
|---|---|
| Main Authors | , , , , , , , , , , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
London
Nature Publishing Group UK
01.09.2012
Nature Publishing Group Springer Nature |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2–7 sequential design–evaluation–redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies. |
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| AbstractList | Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design- evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO- based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies. Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design- evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO- based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies. Leukemia (2012) 26, 1908-1975; doi :10.1038/leu.2012.120 Keywords: EuroFlow; antibody panel; lymphoma; flow cytometry; 8-color immunostaining; standardization; hematological malignancies Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies.Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies. |
| Audience | Academic |
| Author | Lucio, P Böttcher, S Szczepański, T Martin-Ayuso, M van der Velden, V H J Langerak, A W Mejstrikova, E Almeida, J Orfao, A Hrusak, O van Dongen, J J M Mendonça, A Vidriales, M B Flores-Montero, J Lécrevisse, Q Brüggemann, M Lhermitte, L Asnafi, V Rawstron, A de Tute, R Macintyre, E Sedek, L Kalina, T Cullen, M |
| Author_xml | – sequence: 1 givenname: J J M surname: van Dongen fullname: van Dongen, J J M email: j.j.m.vandongen@erasmusmc.nl organization: Department of Immunology, Erasmus MC, University Medical Center Rotterdam (Erasmus MC) – sequence: 2 givenname: L surname: Lhermitte fullname: Lhermitte, L organization: Department of Hematology, Hôpital Necker-Enfants-Malades (AP-HP) and UMR CNRS 8147, University of Paris Descartes – sequence: 3 givenname: S surname: Böttcher fullname: Böttcher, S organization: Second Department of Medicine, University Hospital of Schleswig Holstein, Campus Kiel (UNIKIEL) – sequence: 4 givenname: J surname: Almeida fullname: Almeida, J organization: Department of Medicine and Cytometry Service, Cancer Research Center (IBMCC-CSIC), University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL) – sequence: 5 givenname: V H J surname: van der Velden fullname: van der Velden, V H J organization: Department of Immunology, Erasmus MC, University Medical Center Rotterdam (Erasmus MC) – sequence: 6 givenname: J surname: Flores-Montero fullname: Flores-Montero, J organization: Department of Medicine and Cytometry Service, Cancer Research Center (IBMCC-CSIC), University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL) – sequence: 7 givenname: A surname: Rawstron fullname: Rawstron, A organization: Haematological Malignancy Diagnostic Service (HMDS), University of Leeds (UNIVLEEDS) – sequence: 8 givenname: V surname: Asnafi fullname: Asnafi, V organization: Department of Hematology, Hôpital Necker-Enfants-Malades (AP-HP) and UMR CNRS 8147, University of Paris Descartes – sequence: 9 givenname: Q surname: Lécrevisse fullname: Lécrevisse, Q organization: Department of Medicine and Cytometry Service, Cancer Research Center (IBMCC-CSIC), University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL) – sequence: 10 givenname: P surname: Lucio fullname: Lucio, P organization: Department of Hematology, Portuguese Institute of Oncology (IPOLFG) – sequence: 11 givenname: E surname: Mejstrikova fullname: Mejstrikova, E organization: Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) – sequence: 12 givenname: T surname: Szczepański fullname: Szczepański, T organization: Department of Pediatric Hematology and Oncology, Medical University of Silesia (SUM) – sequence: 13 givenname: T surname: Kalina fullname: Kalina, T organization: Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) – sequence: 14 givenname: R surname: de Tute fullname: de Tute, R organization: Haematological Malignancy Diagnostic Service (HMDS), University of Leeds (UNIVLEEDS) – sequence: 15 givenname: M surname: Brüggemann fullname: Brüggemann, M organization: Second Department of Medicine, University Hospital of Schleswig Holstein, Campus Kiel (UNIKIEL) – sequence: 16 givenname: L surname: Sedek fullname: Sedek, L organization: Department of Pediatric Hematology and Oncology, Medical University of Silesia (SUM) – sequence: 17 givenname: M surname: Cullen fullname: Cullen, M organization: Haematological Malignancy Diagnostic Service (HMDS), University of Leeds (UNIVLEEDS) – sequence: 18 givenname: A W surname: Langerak fullname: Langerak, A W organization: Department of Immunology, Erasmus MC, University Medical Center Rotterdam (Erasmus MC) – sequence: 19 givenname: A surname: Mendonça fullname: Mendonça, A organization: Department of Hematology, Portuguese Institute of Oncology (IPOLFG) – sequence: 20 givenname: E surname: Macintyre fullname: Macintyre, E organization: Department of Hematology, Hôpital Necker-Enfants-Malades (AP-HP) and UMR CNRS 8147, University of Paris Descartes – sequence: 21 givenname: M surname: Martin-Ayuso fullname: Martin-Ayuso, M organization: Cytognos SL – sequence: 22 givenname: O surname: Hrusak fullname: Hrusak, O organization: Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University (DPH/O) – sequence: 23 givenname: M B surname: Vidriales fullname: Vidriales, M B organization: Department of Hematology, University Hospital Salamanca (HUS) and IBSAL – sequence: 24 givenname: A surname: Orfao fullname: Orfao, A organization: Department of Medicine and Cytometry Service, Cancer Research Center (IBMCC-CSIC), University of Salamanca (USAL) and Institute of Biomedical Research of Salamanca (IBSAL) |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/22552007$$D View this record in MEDLINE/PubMed https://hal.science/hal-00740419$$DView record in HAL |
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| Snippet | Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the... |
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| Title | EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes |
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