Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 161-169 |
| Number of pages | 9 |
| Journal | Cancer Immunology, Immunotherapy |
| Volume | 65 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 1 2016 |
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Keywords
- Flow cytometry
- Myeloid-derived suppressor cells
- Phenotyping
- Proficiency panel
ASJC Scopus subject areas
- Cancer Research
- Oncology
- Immunology
- Immunology and Allergy
Cite this
Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry : results from an interim study. / Mandruzzato, Susanna; Brandau, Sven; Britten, Cedrik M.; Bronte, Vincenzo; Damuzzo, Vera; Gouttefangeas, Cécile; Maurer, Dominik; Ottensmeier, Christian; van der Burg, Sjoerd H.; Welters, Marij J P; Walter, Steffen.
In: Cancer Immunology, Immunotherapy, Vol. 65, No. 2, 01.02.2016, p. 161-169.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry
T2 - results from an interim study
AU - Mandruzzato, Susanna
AU - Brandau, Sven
AU - Britten, Cedrik M.
AU - Bronte, Vincenzo
AU - Damuzzo, Vera
AU - Gouttefangeas, Cécile
AU - Maurer, Dominik
AU - Ottensmeier, Christian
AU - van der Burg, Sjoerd H.
AU - Welters, Marij J P
AU - Walter, Steffen
PY - 2016/2/1
Y1 - 2016/2/1
N2 - 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.
AB - 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.
KW - Flow cytometry
KW - Myeloid-derived suppressor cells
KW - Phenotyping
KW - Proficiency panel
UR - http://www.scopus.com/inward/record.url?scp=84955654158&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84955654158&partnerID=8YFLogxK
U2 - 10.1007/s00262-015-1782-5
DO - 10.1007/s00262-015-1782-5
M3 - Article
AN - SCOPUS:84955654158
VL - 65
SP - 161
EP - 169
JO - Cancer Immunology and Immunotherapy
JF - Cancer Immunology and Immunotherapy
SN - 0340-7004
IS - 2
ER -