EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes

被引：620

作者：

van Dongen, J. J. M. ^{[1
]}

Lhermitte, L. ^{[2
,3
]}

Boettcher, S. ^{[4
]}

Almeida, J. ^{[5
,6
,14
]}

van der Velden, V. H. J. ^{[1
]}

Flores-Montero, J. ^{[5
,6
,14
]}

Rawstron, A. ^{[7
]}

Asnafi, V. ^{[2
,3
]}

Lecrevisse, Q. ^{[5
,6
,14
]}

Lucio, P. ^{[8
]}

Mejstrikova, E. ^{[9
]}

Szczepanski, T. ^{[10
]}

Kalina, T. ^{[9
]}

de Tute, R. ^{[7
]}

Brueggemann, M. ^{[4
]}

Sedek, L. ^{[10
]}

Cullen, M. ^{[7
]}

Langerak, A. W. ^{[1
]}

Mendonca, A. ^{[8
]}

Macintyre, E. ^{[2
,3
]}

Martin-Ayuso, M. ^{[11
]}

Hrusak, O. ^{[9
]}

Vidriales, M. B. ^{[12
,13
]}

Orfao, A. ^{[5
,6
,14
]}

机构：

[1] Univ Med Ctr Rotterdam, Erasmus MC, Dept Immunol, NL-3015 GE Rotterdam, Netherlands

[2] Univ Paris 05, Hop Necker Enfants Malades, AP HP, Dept Hematol, Paris, France

[3] Univ Paris 05, UMR CNRS 8147, Paris, France

[4] Univ Hosp Schleswig Holstein, Dept Med 2, Kiel, Germany

[5] Univ Salamanca USAL, Dept Med, Canc Res Ctr IBMCC CSIC, Salamanca, Spain

[6] Inst Biomed Res Salamanca IBSAL, Salamanca, Spain

[7] Univ Leeds UNIVLEEDS, HMDS, Leeds, W Yorkshire, England

[8] Portuguese Inst Oncol IPOLFG, Dept Hematol, Lisbon, Portugal

[9] Charles Univ DPH O, Fac Med 2, Dept Pediat Hematol & Oncol, Prague, Czech Republic

[10] Med Univ Silesia SUM, Dept Pediat Hematol & Oncol, Zabrze, Poland

[11] Cytognos SL, Salamanca, Spain

[12] Univ Hosp Salamanca HUS, Dept Hematol, Salamanca, Spain

[13] IBSAL, Salamanca, Spain

[14] Univ Salamanca USAL, Cytometry Serv, Salamanca, Spain

来源：

LEUKEMIA | 2012年 / 26卷 / 09期

关键词：

EuroFlow; antibody panel; lymphoma; flow cytometry; 8-color immunostaining; standardization; hematological malignancies; ACUTE LYMPHOBLASTIC-LEUKEMIA; MINIMAL RESIDUAL DISEASE; PERIPHERAL T-CELL; CHRONIC LYMPHOPROLIFERATIVE DISORDERS; ACUTE MYELOID-LEUKEMIA; NON-HODGKINS-LYMPHOMA; INTERNATIONAL CONSENSUS RECOMMENDATIONS; NERVOUS-SYSTEM INVOLVEMENT; POLYMERASE-CHAIN-REACTION; V-BETA REPERTOIRE;

D O I：

10.1038/leu.2012.120

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

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.

引用

页码：1908 / 1975

页数：68

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