Harmonemia: a universal strategy for flow cytometry immunophenotyping—A European LeukemiaNet WP10 study

Lacombe, Francis; Bernal, Elisenda; Bloxham, David; Couzens, Stephen; Porta, Matteo Giovanni Della; Johansson, Ulrika; Kern, Wolfgang; Macey, Marion G.; Matthes, Thomas; Morilla, Ricardo; Paiva, Artur; Palacio, Carles; Preijers, Frank; Ratei, Richard; Siitonen, Sanna; Allou, Kaoutar; Porwit, Anna; Béné, Marie C.

doi:10.1038/leu.2016.44

Cited by 43 publications

(64 citation statements)

References 9 publications

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“…Different consortia listed below aimed to address diverse levels of reproducibility: (a) the reproducibility of interpretation (all consortia; some argue that more is not necessary for their purpose such as Harmonemia (17), AIEOP-BFM group (3,(18)(19)(20)(21)(22)(23), and ERIC (21-23)), (b) the reproducibility of enumeration (prominently in immunology: The ONE Study (24,25), HIPC (14,26,27), but also in the leukemia residual disease detection: EuroFlow (28)(29)(30), COG group (31), and ERIC (21,22)), (c) the reproducibility of staining pattern is typically achieved when the same panel of reagents is used (The ONE Study (25), HIPC (26), EuroFlow (32,33), and COG (31)), which allows definition of the manual analysis strategy and also automated analysis tools, and (d) the reproducibility of patterns including the staining intensity that allows analysis standardization and database-assisted comparison to previous cases in interlaboratory settings: EuroFlow (34) or The Canadian National Transplant Research Program that analyzed multiple immune cells in multiple centers by automated analysis (35). Different consortia listed below aimed to address diverse levels of reproducibility: (a) the reproducibility of interpretation (all consortia; some argue that more is not necessary for their purpose such as Harmonemia (17), AIEOP-BFM group (3,(18)(19)(20)(21)(22)(23), and ERIC (21-23)), (b) the reproducibility of enumeration (prominently in immunology: The ONE Study (24,25), HIPC (14,2...…”

Section: Existing Efforts To Achieve Reproducibilitymentioning

confidence: 99%

Reproducibility of Flow Cytometry Through Standardization: Opportunities and Challenges

Kalina

2019

Cytometry Pt A

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There is an agreement in the field that interlaboratory reproducibility of flow cytometry measurements as well as the whole studies might be improved by a consensual use of methodological approach. Typically, a consensus is made on a crucial markers needed in the immunostaining panel, sometimes on the particular fluorochrome conjugates and rarely on a complete set of methods for sample preparation. The term "standardization" is used to describe the complete set of methodical steps, while "harmonization" is used for partial agreement on the method. Standardization can provide a platform for improved reproducibility of cytometry results over prolonged periods of time, across different sites and across different instruments. For the purpose of structured discussion, several desired aims are described: common interpretation of the immunophenotype definition of a target subset, accurate quantification, reproducible pattern of a multicolor immunophenotype, and reproducible intensity of all measured parameters. An overview of how standardization was approached by several large consortia is provided: EuroFlow, The ONE Study, Human Immunology Project Consortium (HIPC), and several other groups. Their particular aims and the tools adopted to reach those aims are noted. How those standardization efforts were adopted in the field and how the resulting outcome was evaluated is reviewed. Multiple challenges in the instrument hardware design, instrument setup tools, reagent design, and quality features need to be addressed to achieve optimal standardization. Furthermore, the aims of different studies vary, and thus, the reasonable requirements for standardization differ. A framework of reference for the reasonable outcomes of different approaches is offered. Finally, it is argued that complete standardization is important not only for the reproducibility of measurements but also for education, for quality assessment and for algorithmic data analysis. The different standardized approaches can and in fact should serve as benchmarking reference tools for the development of future flow cytometry studies.

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Section: Existing Efforts To Achieve Reproducibilitymentioning

confidence: 99%

Reproducibility of Flow Cytometry Through Standardization: Opportunities and Challenges

Kalina

2019

Cytometry Pt A

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“…A concept proposed by the Harmonemia group was adopted. The target channels (TC) were established at UHN using FlowSet Pro beads (BC, ref.…”

Section: Methodsmentioning

confidence: 99%

“…The new panel should also provide orientation in T and NK cell subsets even in paucicellular samples such as cerebrospinal fluids (CSF) and FNA. We applied the Harmonemia group′s recently proposed protocol of harmonization of instrument settings to implement the newly developed LPD‐ST in two other laboratories. Additionally, the novel technology of dried monoclonal antibody reagents was applied in the clinical application of LPD‐ST.…”

Section: Introductionmentioning

confidence: 99%

Ten‐color 15‐antibody flow cytometry panel for immunophenotyping of lymphocyte population

Rajab¹,

Axler

Leung

et al. 2017

Int J Lab Hematology

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Summary We have developed a lymphoproliferative disorder screening tube (LPD‐ST) with the aim to provide comprehensive immunophenotyping of lymphocyte subsets with minimal need for additional testing. The LPD‐ST consists of CD4/kappa FITC, CD8/lambda PE, CD3/CD14ECD, CD38PC5.5, CD20/CD56PC7, CD10APC, CD19APC‐A700, CD5APC‐A750, CD57/CD23PB and CD45KO. The LPD‐ST was validated against previously used lymphocyte subset panels in Canada (n=60) and in Sweden (n=43) and against the OneFlow™ LST (n=60). The LPD‐ST panel was then implemented in clinical practice using dried monoclonal antibody reagents (Duraclone®) on 649 patient samples in Sweden. In 204 of 649 samples (31%), a monotypic B‐cell population was found. Of these cases, a final diagnosis could be rendered in 106 cases (52%), and in the remainder, additional B‐cell immunophenotyping was performed. In 20 (3%) samples, an aberrant T‐cell population was confirmed by additional testing. Of 425 samples diagnosed as normal/reactive lymphoid tissue, 50 (12%) required additional immunophenotyping, mostly due to an abnormal CD4/CD8 ratio. The LPD‐ST tube significantly minimizes the need for additional testing, improves the turn‐around time, and reduces the cost of LPD immunophenotyping. It is also suitable for investigating paucicellular samples such as cerebrospinal fluid or fine needle aspirates.

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“…With the increasing number of participants from the Americas and Asia, the ELN has evolved from a European to an international network. Some of the more recent ELN recommendations are listed in Table …”

Section: Recommendations and Guidelines: 2013 To 2017mentioning

confidence: 99%

Advancing a field by building consortia: The example of the European LeukemiaNet

Hehlmann

2018

Cancer

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Harmonemia: a universal strategy for flow cytometry immunophenotyping—A European LeukemiaNet WP10 study

Cited by 43 publications

References 9 publications

Reproducibility of Flow Cytometry Through Standardization: Opportunities and Challenges

Reproducibility of Flow Cytometry Through Standardization: Opportunities and Challenges

Ten‐color 15‐antibody flow cytometry panel for immunophenotyping of lymphocyte population

Advancing a field by building consortia: The example of the European LeukemiaNet

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