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Title:
Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study | Cancer Immunology, Immunotherapy
Description:
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.
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mdsc, cells, cancer, subsets, pubmed, article, panel, mdscs, gating, google, scholar, cas, variance, cell, suppressor, results, laboratories, data, human, university, myeloidderived, proficiency, participants, patients, fig, immunol, strategy, myeloid, color, central, analysis, phenotyping, cytometry, number, flow, mandruzzato, identification, germany, marker, high, granulocytic, identified, markers, staining, panels, usa, britten, interlaboratory, immature, lymphocytes,
Topics {âïž}
mdsc7 cd15â/cd14+/cd33high/hla-drlow [18] mdsc7 cd15â/cd14+/cd33high/hla-drlow/â [18] myeloid-derived suppressor cells mdsc10 linâ/hla-drlow/cd11b+ [19] mdsc10 linâ/hla-drlow/â/cd11b+ [19] granulocyte-derived hydrogen peroxide cd11b+/gr-1high/ly6câ/ly6ghigh cd11b+/gr-1int/ly6chigh/ly6gâ [6] mdsc3 linâ/hla-drâ/cd33+ [10] mdsc3 linâ/hla-drâ/cd33+ [10 article download pdf myeloid suppressor cells mdsc4 cd14+/hla-drlow/â [11] hla-peptide multimer assay mdsc6 cd15+/fsclow/sschigh [13] wilcoxon signed-rank test wilcoxon matched-pair test myeloid cell-mediated immunosuppression mdsc9 cd14â/cd15â/cd33high immatics biotechnologies gmbh van der burg & marij single-dose cyclophosphamide associates high inter-laboratory variance reports inter-laboratory variance multimer-based proficiency panel high inter-center variance high inter-laboratory variation inter-laboratory variability arises calculate intra-laboratory variance mdsc5 cd15+/cd14â/cd11b+ [15] diaz-montero cm immature myeloid cells van der burg cd33 positive cells colony-stimulating factors renal cell carcinoma privacy choices/manage cookies immune suppression mediated highly fluorescent cells dimly fluorescent cells cd11b+/gr-1+ cells hla-multimer staining phenotypic definition mdsc8 cd15+/cd33high cancer vaccine ima901 inter-laboratory variance data analysis revealed dead-cell marker multipeptide immune response proficiency panels based
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headline:Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study
description: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.
datePublished:2016-01-04T00:00:00Z
dateModified:2016-01-04T00:00:00Z
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Myeloid-derived suppressor cells
Phenotyping
Proficiency panel
Flow cytometry
Oncology
Immunology
Cancer Research
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headline:Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study
description: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.
datePublished:2016-01-04T00:00:00Z
dateModified:2016-01-04T00:00:00Z
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Myeloid-derived suppressor cells
Phenotyping
Proficiency panel
Flow cytometry
Oncology
Immunology
Cancer Research
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