G-CSF TREATMENT IN VIVO INDUCES BONE MARROW RELEASE OF PD-L1 POSITIVE MYELOID DERIVED SUPPRESSIVE CELLS.
(Abstract release date: 05/19/16)
EHA Library. Lereclus E. 06/09/16; 132684; E1135
Dr. Emilie Lereclus
Contributions
Contributions
Abstract
Abstract: E1135
Type: Eposter Presentation
Background
For its two main adverse effects which engage the vital prognosis, graft versus host disease (GVHD) and graft versus leukemia (GVL), tuning the immune system is the major challenge for successful cure of the patient who benefits from Hematopoietic Stem Cell (HSC) transplantation. Myeloid derived suppressor cells (MDSCs) are supposed to be an immature leukocyte subset composed of granulocytic cells (GrMDSC) and monocytic cells (MoMDSC). They are well described in different pathologies such as cancer, burns and sepsis. Few publications suggest that G-CSF induces bone marrow release of myeloid derived suppressive cells (MDSCs). But, phenotypically, it is hardly possible to distinguish them from other myeloid precursors with the classical lineage markers. Recently, immunosuppressive action of MDSCs was reported to be due to the PD-L1 molecule that renders these cells able to kill T-cells through the PD-1/ PD-L1 axis.
Aims
Using PD-L1 as an additional marker, the aim of this study was to quantify MDSCs subsets (GrMDSC and MoMDSC) before and after G-CSF treatment as well as in bone marrow of donors prior Hematopoietic Stem Cell (HSC) transplantation.
Methods
Recruitment of donors was done in two hospitals university centers (Bordeaux and Limoges, France) .We analyzed 10 samples of peripheral blood (PB) before and 17 after G-CSF treatment, 27 samples of cells collected after apheresis (PSC) and 5 samples of bone marrow (BM) as control. We measured the rate of GrMDSC (CD45+ Lin- CD11b+ CD14- CD33+ CD16- HLA-DR-PD-L1+) and MoMDSC (CD45+ CD11b+ CD14+ CD33+ CD16- HLA-DR- PD-L1+) in BM, PB and PSC sample by flow cytometry. Percentage and absolute counts CD34 and CD3 positive cells in G-CSF treated donors have been also collected. Differences between groups were tested for significance with the Student t-test.
Results
White blood cell count was 5.09 +/- 2.55 G/L and 45.77 +/- 25.5 G/L before and after G-CSF treatment. Levels of PD-L1+ HLA DR- MoMDSCs and CD33+ Lin- PDL1+ GrMDSCs were 0.0013 +/- 0.0027 G/L (0.023% +/- 0.042%) and 0.058 +/-0.048 G/L (1.15% +/- 0.91%) in PB before and were 0.078 +/- 0.085 G/L (0.21% +/- 0.25%) and 12.31 +/- 6.52 G/L (28% +/- 13.3%) after G-CSF treatment (p=0.002 and <10-4 respectively). Percentages of MoMDSCs and GrMDSCs with PDL1 expression in bone marrow were 0.0033% +/- 0.0071% and 0.84% +/- 1.21%. Apheresis treatment was associated with enrichment in PDL1+ MoMDCs but with a loss of PDL1+GrMDSCs. Nevertheless, when compared to bone marrow, percentages and total numbers of both PDL1+ MoMDSC and GrMDSC in apheresis products were still much higher than in bone marrow (p<0.0001 and p=0.0006). There was no correlation between levels of CD34+ stem cells, CD3+ or CD19+ lymphocytes and MDSCs after G-CSF treatment.
Conclusion
In this study we have shown that PDL1+ MDSCs were virtually absent in normal bone marrow and that G-CSF treatment induced a strong MDSC production. We have also demonstrated that, compared to PB, the product of apheresis was even more enriched in MoMDSC but was partially depleted in GrMDSC. These results point a major difference between bone marrow HSCs and PSCs in terms of immunomodulation capacities of the engraftment product.
Session topic: E-poster
Keyword(s): G-CSF, Myeloid differentiation
Type: Eposter Presentation
Background
For its two main adverse effects which engage the vital prognosis, graft versus host disease (GVHD) and graft versus leukemia (GVL), tuning the immune system is the major challenge for successful cure of the patient who benefits from Hematopoietic Stem Cell (HSC) transplantation. Myeloid derived suppressor cells (MDSCs) are supposed to be an immature leukocyte subset composed of granulocytic cells (GrMDSC) and monocytic cells (MoMDSC). They are well described in different pathologies such as cancer, burns and sepsis. Few publications suggest that G-CSF induces bone marrow release of myeloid derived suppressive cells (MDSCs). But, phenotypically, it is hardly possible to distinguish them from other myeloid precursors with the classical lineage markers. Recently, immunosuppressive action of MDSCs was reported to be due to the PD-L1 molecule that renders these cells able to kill T-cells through the PD-1/ PD-L1 axis.
Aims
Using PD-L1 as an additional marker, the aim of this study was to quantify MDSCs subsets (GrMDSC and MoMDSC) before and after G-CSF treatment as well as in bone marrow of donors prior Hematopoietic Stem Cell (HSC) transplantation.
Methods
Recruitment of donors was done in two hospitals university centers (Bordeaux and Limoges, France) .We analyzed 10 samples of peripheral blood (PB) before and 17 after G-CSF treatment, 27 samples of cells collected after apheresis (PSC) and 5 samples of bone marrow (BM) as control. We measured the rate of GrMDSC (CD45+ Lin- CD11b+ CD14- CD33+ CD16- HLA-DR-PD-L1+) and MoMDSC (CD45+ CD11b+ CD14+ CD33+ CD16- HLA-DR- PD-L1+) in BM, PB and PSC sample by flow cytometry. Percentage and absolute counts CD34 and CD3 positive cells in G-CSF treated donors have been also collected. Differences between groups were tested for significance with the Student t-test.
Results
White blood cell count was 5.09 +/- 2.55 G/L and 45.77 +/- 25.5 G/L before and after G-CSF treatment. Levels of PD-L1+ HLA DR- MoMDSCs and CD33+ Lin- PDL1+ GrMDSCs were 0.0013 +/- 0.0027 G/L (0.023% +/- 0.042%) and 0.058 +/-0.048 G/L (1.15% +/- 0.91%) in PB before and were 0.078 +/- 0.085 G/L (0.21% +/- 0.25%) and 12.31 +/- 6.52 G/L (28% +/- 13.3%) after G-CSF treatment (p=0.002 and <10-4 respectively). Percentages of MoMDSCs and GrMDSCs with PDL1 expression in bone marrow were 0.0033% +/- 0.0071% and 0.84% +/- 1.21%. Apheresis treatment was associated with enrichment in PDL1+ MoMDCs but with a loss of PDL1+GrMDSCs. Nevertheless, when compared to bone marrow, percentages and total numbers of both PDL1+ MoMDSC and GrMDSC in apheresis products were still much higher than in bone marrow (p<0.0001 and p=0.0006). There was no correlation between levels of CD34+ stem cells, CD3+ or CD19+ lymphocytes and MDSCs after G-CSF treatment.
Conclusion
In this study we have shown that PDL1+ MDSCs were virtually absent in normal bone marrow and that G-CSF treatment induced a strong MDSC production. We have also demonstrated that, compared to PB, the product of apheresis was even more enriched in MoMDSC but was partially depleted in GrMDSC. These results point a major difference between bone marrow HSCs and PSCs in terms of immunomodulation capacities of the engraftment product.
Session topic: E-poster
Keyword(s): G-CSF, Myeloid differentiation
Abstract: E1135
Type: Eposter Presentation
Background
For its two main adverse effects which engage the vital prognosis, graft versus host disease (GVHD) and graft versus leukemia (GVL), tuning the immune system is the major challenge for successful cure of the patient who benefits from Hematopoietic Stem Cell (HSC) transplantation. Myeloid derived suppressor cells (MDSCs) are supposed to be an immature leukocyte subset composed of granulocytic cells (GrMDSC) and monocytic cells (MoMDSC). They are well described in different pathologies such as cancer, burns and sepsis. Few publications suggest that G-CSF induces bone marrow release of myeloid derived suppressive cells (MDSCs). But, phenotypically, it is hardly possible to distinguish them from other myeloid precursors with the classical lineage markers. Recently, immunosuppressive action of MDSCs was reported to be due to the PD-L1 molecule that renders these cells able to kill T-cells through the PD-1/ PD-L1 axis.
Aims
Using PD-L1 as an additional marker, the aim of this study was to quantify MDSCs subsets (GrMDSC and MoMDSC) before and after G-CSF treatment as well as in bone marrow of donors prior Hematopoietic Stem Cell (HSC) transplantation.
Methods
Recruitment of donors was done in two hospitals university centers (Bordeaux and Limoges, France) .We analyzed 10 samples of peripheral blood (PB) before and 17 after G-CSF treatment, 27 samples of cells collected after apheresis (PSC) and 5 samples of bone marrow (BM) as control. We measured the rate of GrMDSC (CD45+ Lin- CD11b+ CD14- CD33+ CD16- HLA-DR-PD-L1+) and MoMDSC (CD45+ CD11b+ CD14+ CD33+ CD16- HLA-DR- PD-L1+) in BM, PB and PSC sample by flow cytometry. Percentage and absolute counts CD34 and CD3 positive cells in G-CSF treated donors have been also collected. Differences between groups were tested for significance with the Student t-test.
Results
White blood cell count was 5.09 +/- 2.55 G/L and 45.77 +/- 25.5 G/L before and after G-CSF treatment. Levels of PD-L1+ HLA DR- MoMDSCs and CD33+ Lin- PDL1+ GrMDSCs were 0.0013 +/- 0.0027 G/L (0.023% +/- 0.042%) and 0.058 +/-0.048 G/L (1.15% +/- 0.91%) in PB before and were 0.078 +/- 0.085 G/L (0.21% +/- 0.25%) and 12.31 +/- 6.52 G/L (28% +/- 13.3%) after G-CSF treatment (p=0.002 and <10-4 respectively). Percentages of MoMDSCs and GrMDSCs with PDL1 expression in bone marrow were 0.0033% +/- 0.0071% and 0.84% +/- 1.21%. Apheresis treatment was associated with enrichment in PDL1+ MoMDCs but with a loss of PDL1+GrMDSCs. Nevertheless, when compared to bone marrow, percentages and total numbers of both PDL1+ MoMDSC and GrMDSC in apheresis products were still much higher than in bone marrow (p<0.0001 and p=0.0006). There was no correlation between levels of CD34+ stem cells, CD3+ or CD19+ lymphocytes and MDSCs after G-CSF treatment.
Conclusion
In this study we have shown that PDL1+ MDSCs were virtually absent in normal bone marrow and that G-CSF treatment induced a strong MDSC production. We have also demonstrated that, compared to PB, the product of apheresis was even more enriched in MoMDSC but was partially depleted in GrMDSC. These results point a major difference between bone marrow HSCs and PSCs in terms of immunomodulation capacities of the engraftment product.
Session topic: E-poster
Keyword(s): G-CSF, Myeloid differentiation
Type: Eposter Presentation
Background
For its two main adverse effects which engage the vital prognosis, graft versus host disease (GVHD) and graft versus leukemia (GVL), tuning the immune system is the major challenge for successful cure of the patient who benefits from Hematopoietic Stem Cell (HSC) transplantation. Myeloid derived suppressor cells (MDSCs) are supposed to be an immature leukocyte subset composed of granulocytic cells (GrMDSC) and monocytic cells (MoMDSC). They are well described in different pathologies such as cancer, burns and sepsis. Few publications suggest that G-CSF induces bone marrow release of myeloid derived suppressive cells (MDSCs). But, phenotypically, it is hardly possible to distinguish them from other myeloid precursors with the classical lineage markers. Recently, immunosuppressive action of MDSCs was reported to be due to the PD-L1 molecule that renders these cells able to kill T-cells through the PD-1/ PD-L1 axis.
Aims
Using PD-L1 as an additional marker, the aim of this study was to quantify MDSCs subsets (GrMDSC and MoMDSC) before and after G-CSF treatment as well as in bone marrow of donors prior Hematopoietic Stem Cell (HSC) transplantation.
Methods
Recruitment of donors was done in two hospitals university centers (Bordeaux and Limoges, France) .We analyzed 10 samples of peripheral blood (PB) before and 17 after G-CSF treatment, 27 samples of cells collected after apheresis (PSC) and 5 samples of bone marrow (BM) as control. We measured the rate of GrMDSC (CD45+ Lin- CD11b+ CD14- CD33+ CD16- HLA-DR-PD-L1+) and MoMDSC (CD45+ CD11b+ CD14+ CD33+ CD16- HLA-DR- PD-L1+) in BM, PB and PSC sample by flow cytometry. Percentage and absolute counts CD34 and CD3 positive cells in G-CSF treated donors have been also collected. Differences between groups were tested for significance with the Student t-test.
Results
White blood cell count was 5.09 +/- 2.55 G/L and 45.77 +/- 25.5 G/L before and after G-CSF treatment. Levels of PD-L1+ HLA DR- MoMDSCs and CD33+ Lin- PDL1+ GrMDSCs were 0.0013 +/- 0.0027 G/L (0.023% +/- 0.042%) and 0.058 +/-0.048 G/L (1.15% +/- 0.91%) in PB before and were 0.078 +/- 0.085 G/L (0.21% +/- 0.25%) and 12.31 +/- 6.52 G/L (28% +/- 13.3%) after G-CSF treatment (p=0.002 and <10-4 respectively). Percentages of MoMDSCs and GrMDSCs with PDL1 expression in bone marrow were 0.0033% +/- 0.0071% and 0.84% +/- 1.21%. Apheresis treatment was associated with enrichment in PDL1+ MoMDCs but with a loss of PDL1+GrMDSCs. Nevertheless, when compared to bone marrow, percentages and total numbers of both PDL1+ MoMDSC and GrMDSC in apheresis products were still much higher than in bone marrow (p<0.0001 and p=0.0006). There was no correlation between levels of CD34+ stem cells, CD3+ or CD19+ lymphocytes and MDSCs after G-CSF treatment.
Conclusion
In this study we have shown that PDL1+ MDSCs were virtually absent in normal bone marrow and that G-CSF treatment induced a strong MDSC production. We have also demonstrated that, compared to PB, the product of apheresis was even more enriched in MoMDSC but was partially depleted in GrMDSC. These results point a major difference between bone marrow HSCs and PSCs in terms of immunomodulation capacities of the engraftment product.
Session topic: E-poster
Keyword(s): G-CSF, Myeloid differentiation
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