The MDSC Detection Kit is developed to allow standardization and simplification of flow cytometric analysis of the subpopulations of MDSCs described so far. For greater flexibility, the product configuration also allows the extension of the analysis panel with additional specificities of interest (e.g. CD10, CD66b, CD15, and LOX-1).

Data and images for MDSC Detection Kit, human

Figures

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
A) Gating strategy:
All events
Gated on R1
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
Gated on R2
Gated on R3
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
B) Identification of low density PMN cells (PMN-MDSCs):
Gated on R4
Gated on R5
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
C) Identification of M-MDSCs:
Gated on R6
Gated on R6
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
D) Identification of e-MDSCs:
Gated on R7
Gated on R7
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.
View details

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant
®
Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45
+
cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD
cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter
high
(SSC
high
) cells, which correspond to low density PMN-MDSCs (region R5), CD14
+
cells (region R6) and SSC
low
CD14
cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14
+
HLA-DR
. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing
CD33
int
.

Specifications for MDSC Detection Kit, human

Overview

The MDSC Detection Kit is developed to allow standardization and simplification of flow cytometric analysis of the subpopulations of MDSCs described so far. For greater flexibility, the product configuration also allows the extension of the analysis panel with additional specificities of interest (e.g. CD10, CD66b, CD15, and LOX-1).

Detailed product information

Background information

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of mononuclear and polymorphonuclear myeloid cells with different immunosuppressive as well as disease promoting functions¹,². In healthy volunteers, they are found in very small numbers, but can expand considerably in pathological conditions. Depending on the type and stage of the disease, the MDSC population might be present in different maturation phases. In human, three major MDSC subsets can be identified: low density polymorphonuclear-MDSCs (PMN-MDSCs), also known as granulocytic MDSCs (G-MDSCs), monocytic- MDSCs (M-MDSCs), and early-MDSCs (e-MDSCs)³. Validated unambiguous phenotypic markers for MDSCs are still lacking and therefore various combinations of specificities are used for their identification⁴.

Applications

Detection and quantification of MDSC subsets in human PBMCs. This product might be used for analysis of other starting materials, such as whole blood, lysed whole blood, or single-cell suspensions from human tissues. For such applications, optional reagents might be required, such as Red Blood Cell Lysis Solution (10×) or additional fluorochrome-conjugated antibodies.

References for MDSC Detection Kit, human

Publications

  1. Marigo, I. et al. (2010) Tumor-induced tolerance and immune suppression depend on the C/EBPβ transcription factor. Immunity 32(6): 790-802
  2. Marini, O. et al. (2017)
    Mature CD10
    +
    and immature CD10
    neutrophils present in G-CSF–treated donors display opposite effects on T cells.
    Blood 129(10): 1343-1356
  3. Veglia, F. et al. (2018) Myeloid-derived suppressor cells coming of age. Nat. Immunol. 19(2): 108-119
  4. Mandruzzato, S. et al. (2016) Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study. Cancer Immunol. Immunother. 65(2): 161-169
  5. Gabrilovich, D. I. et al. (2012) Coordinated regulation of myeloid cells by tumours. Nat. Rev. Immunol. 12(4): 253-268
  6. Bruger, A. M. et al. (2020) Protocol to assess the suppression of T-cell proliferation by human MDSC. Meth. Enzymol. 632: 155-192
  7. Lang, S. et al. (2018) Clinical relevance and suppressive capacity of human myeloid-derived suppressor cell subsets. Clin Cancer Res. 24(19): 4834-4844
  8. Bronte, V. et al. (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun. 7(1): 12150
  9. Condamine, T. et al. (2016) Lectin-type oxidized LDL receptor-1 distinguishes population of human polymorphonuclear myeloid-derived suppressor cells in cancer patients. Sci Immunol 1(2): pii:aaf8943

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