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Cancer Immunol Immunother. 2016 Feb;65(2):161-9. doi: 10.1007/s00262-015-1782-5. Epub 2016 Jan 4.

Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study.

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Section of Oncology and Immunology, Department of Surgery, Oncology and Gastroenterology, University of Padova, Via Gattamelata, 64, 35128, Padua, Italy.
Veneto Institute of Oncology IOV - IRCCS, Padua, Italy.
Department of Otorhinolaryngology, University Hospital Essen, Essen, Germany.
TRON Translationale Onkologie an der Universitätsmedizin der Johannes Gutenberg-Universität Mainz GmbH, Mainz, Germany.
Cell Therapy Group, Immuno-Oncology and Combinations, GlaxoSmithKline, Stevenage, UK.
Section of Immunology, Department of Pathology and Diagnostics, Verona University Hospital, Verona, Italy.
Section of Oncology and Immunology, Department of Surgery, Oncology and Gastroenterology, University of Padova, Via Gattamelata, 64, 35128, Padua, Italy.
Veneto Institute of Oncology IOV - IRCCS, Padua, Italy.
Department of Immunology, Institute for Cell Biology, University of Tübingen, Tübingen, Germany.
Immatics Biotechnologies GmbH, Tübingen, Germany.
Cancer Sciences Unit, Faculty of Medicine, Experimental Cancer Medicine Centre, Southampton General Hospital, University of Southampton, Tremona Road, Southampton, UK.
Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.


There is an increasing interest for monitoring circulating myeloid-derived suppressor cells (MDSCs) in cancer patients, but there are also divergences in their phenotypic definition. To overcome this obstacle, the Cancer Immunoguiding Program under the umbrella of the Association of Cancer Immunotherapy is coordinating a proficiency panel program that aims at harmonizing MDSC phenotyping. After a consultation period, a two-stage approach was designed to harmonize MDSC phenotype. In the first step, an international consortium of 23 laboratories immunophenotyped 10 putative MDSC subsets on pretested, peripheral blood mononuclear cells of healthy donors to assess the level of concordance and define robust marker combinations for the identification of circulating MDSCs. At this stage, no mandatory requirements to standardize reagents or protocols were introduced. Data analysis revealed a small intra-laboratory, but very high inter-laboratory variance for all MDSC subsets, especially for the granulocytic subsets. In particular, the use of a dead-cell marker altered significantly the reported percentage of granulocytic MDSCs, confirming that these cells are especially sensitive to cryopreservation and/or thawing. Importantly, the gating strategy was heterogeneous and associated with high inter-center variance. Overall, our results document the high variability in MDSC phenotyping in the multicenter setting if no harmonization/standardization measures are applied. Although the observed variability depended on a number of identified parameters, the main parameter associated with variation was the gating strategy. Based on these findings, we propose further efforts to harmonize marker combinations and gating parameters to identify strategies for a robust enumeration of MDSC subsets.


Flow cytometry; Myeloid-derived suppressor cells; Phenotyping; Proficiency panel

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