DEPLETION OF PROINFLAMMATORY MONOCYTES AND INCREASED LEVELS OF CD56 EXPRESSION ON CLASSICAL MONOCYTES IN THE PERIPHERAL FROM PATIENTS WITH LOWER RISK MYELODYSPLASTIC SYNDROMES
(Abstract release date: 05/19/16)
EHA Library. Cabral R. 06/09/16; 134806; PB1906
Prof. Renata Cabral
Contributions
Contributions
Abstract
Abstract: PB1906
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are hematologic neoplasms characterized by morphologic dysplasia and peripheral blood (PB) cytopenias. Flow cytometry (FC) studies have revealed that bone marrow monocytes (Mon) from MDS patients have abnormal maturation patterns and aberrant antigen expression. However, the immunophenotypic alterations in PB monocytes (PB-Mon) have been much less explored.
Aims
To evaluate the levels of CD11b, CD11c, CD13, CD14, CD15, CD16, CD64 and HLA-DR expression in PB-Mon from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI). To quantify the proinflammatory (CD14+loCD16+) and classical (CD14+hiCD16-) PB-Mon subsets, and to search for abnormal CD56 expression on PB-Mon from LR-MDS patients.
Methods
Fourteen patients with LR-MDS (8 males, median age 76 years), and an equal number of NI (blood donors, 8 males, median age 55 years) were studied. Patients who were being treated with myeloid growth factors were excluded, as did patients with active infections and other concomitant neoplasms. The median time from the diagnosis was 7.6 years, ranging from 0.5 to 12.6 years. Seven patients had refractory anemia with ringed sideroblasts (RARS), 4 patients had refractory cytopenias with unilineage dysplasia (RCUD), and 3 patients had refractory cytopenias with multilineage dysplasia (RCMD). Eight patients had low IPSS risk and 6 patients had intermediate 1 IPSS risk. The median PB-Mon count was of 426/mm3 (17 to 1132), in LR-MDS (>1000/mm3 in 1 case) and 482/mm3 (294 to 937), in NI (no cases with >1000/mm3).PB samples were collected into EDTA-K3 containing tubes. Cell immunophenotyping was performed by 8-color FC using fluorochrome conjugated monoclonal antibodies with different specificities (T1: CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-APC, CD14-APC-H7, CD16-V450, CD45-KO; T2: HLA-DR-FITC, CD64-PE, CD34-PerCPCy5.5, CD56-PC7, CD11c-APC, CD14-APC-H7, CD16-V450, CD45-KO), and a whole blood stain-lyse-and-then wash method (FACSLysing, Becton Dickinson–BD). A normal PB sample was run in parallel with each patient. Sample acquisition was performed in a FACSCanto II flow cytometer (BD), calibrated according to the Euroflow SOP. Data analysis was done with Infinicyt (Cytognos). Results are expressed as median, minimum and maximum values of the median fluorescence intensity observed for each marker. P values < 0.05 were considered statistically significant (Mann-Whitney U test).
Results
PB-Mon from patients with LR-MDS had FSC and SSC similar to the PB-Mon from NI (p>0.05). In the same way, the overall levels of CD13, CD14, CD15, CD45 and CD64 expression on PB-Mon from patients with LR-MDS did not differ significantly from those observed in NI (p>0.05). In contrast, PB-Mon from LR-MDS patients had significantly higher levels of CD56 (p=0.006), and lower levels of CD11c (p=0.004), CD16 (p=0.005) and HLA-DR (p=0.042), and showed a tendency for a lower CD11b expression (p=0.089), as compared to NI. Additionally, PB-Mon from LR-MDS patients had a marked decrease in the fraction of CD14+loCD16+ subset (3.1% ranging from 0.1 to 8.1% and 11.5% ranging from 4.1 to 27.7%, respectively; p<0.001). Moreover, CD14+hiCD16- PB-Mon from LR-MDS patients had a significantly higher % of CD56+ cells, as compared to NI (14.8% ranging from 0.1 to 98.7% and 7.1% ranging from 0.1 to 14.9%, respectively; p=0.026).
Conclusion
PB-Mon from patients with LR-MDS have abnormal levels of CD11b, CD11c, CD16 and CD56 expression and a high classical / proinflammatory Mon subset ratio.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Monocyte
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are hematologic neoplasms characterized by morphologic dysplasia and peripheral blood (PB) cytopenias. Flow cytometry (FC) studies have revealed that bone marrow monocytes (Mon) from MDS patients have abnormal maturation patterns and aberrant antigen expression. However, the immunophenotypic alterations in PB monocytes (PB-Mon) have been much less explored.
Aims
To evaluate the levels of CD11b, CD11c, CD13, CD14, CD15, CD16, CD64 and HLA-DR expression in PB-Mon from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI). To quantify the proinflammatory (CD14+loCD16+) and classical (CD14+hiCD16-) PB-Mon subsets, and to search for abnormal CD56 expression on PB-Mon from LR-MDS patients.
Methods
Fourteen patients with LR-MDS (8 males, median age 76 years), and an equal number of NI (blood donors, 8 males, median age 55 years) were studied. Patients who were being treated with myeloid growth factors were excluded, as did patients with active infections and other concomitant neoplasms. The median time from the diagnosis was 7.6 years, ranging from 0.5 to 12.6 years. Seven patients had refractory anemia with ringed sideroblasts (RARS), 4 patients had refractory cytopenias with unilineage dysplasia (RCUD), and 3 patients had refractory cytopenias with multilineage dysplasia (RCMD). Eight patients had low IPSS risk and 6 patients had intermediate 1 IPSS risk. The median PB-Mon count was of 426/mm3 (17 to 1132), in LR-MDS (>1000/mm3 in 1 case) and 482/mm3 (294 to 937), in NI (no cases with >1000/mm3).PB samples were collected into EDTA-K3 containing tubes. Cell immunophenotyping was performed by 8-color FC using fluorochrome conjugated monoclonal antibodies with different specificities (T1: CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-APC, CD14-APC-H7, CD16-V450, CD45-KO; T2: HLA-DR-FITC, CD64-PE, CD34-PerCPCy5.5, CD56-PC7, CD11c-APC, CD14-APC-H7, CD16-V450, CD45-KO), and a whole blood stain-lyse-and-then wash method (FACSLysing, Becton Dickinson–BD). A normal PB sample was run in parallel with each patient. Sample acquisition was performed in a FACSCanto II flow cytometer (BD), calibrated according to the Euroflow SOP. Data analysis was done with Infinicyt (Cytognos). Results are expressed as median, minimum and maximum values of the median fluorescence intensity observed for each marker. P values < 0.05 were considered statistically significant (Mann-Whitney U test).
Results
PB-Mon from patients with LR-MDS had FSC and SSC similar to the PB-Mon from NI (p>0.05). In the same way, the overall levels of CD13, CD14, CD15, CD45 and CD64 expression on PB-Mon from patients with LR-MDS did not differ significantly from those observed in NI (p>0.05). In contrast, PB-Mon from LR-MDS patients had significantly higher levels of CD56 (p=0.006), and lower levels of CD11c (p=0.004), CD16 (p=0.005) and HLA-DR (p=0.042), and showed a tendency for a lower CD11b expression (p=0.089), as compared to NI. Additionally, PB-Mon from LR-MDS patients had a marked decrease in the fraction of CD14+loCD16+ subset (3.1% ranging from 0.1 to 8.1% and 11.5% ranging from 4.1 to 27.7%, respectively; p<0.001). Moreover, CD14+hiCD16- PB-Mon from LR-MDS patients had a significantly higher % of CD56+ cells, as compared to NI (14.8% ranging from 0.1 to 98.7% and 7.1% ranging from 0.1 to 14.9%, respectively; p=0.026).
Conclusion
PB-Mon from patients with LR-MDS have abnormal levels of CD11b, CD11c, CD16 and CD56 expression and a high classical / proinflammatory Mon subset ratio.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Monocyte
Abstract: PB1906
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are hematologic neoplasms characterized by morphologic dysplasia and peripheral blood (PB) cytopenias. Flow cytometry (FC) studies have revealed that bone marrow monocytes (Mon) from MDS patients have abnormal maturation patterns and aberrant antigen expression. However, the immunophenotypic alterations in PB monocytes (PB-Mon) have been much less explored.
Aims
To evaluate the levels of CD11b, CD11c, CD13, CD14, CD15, CD16, CD64 and HLA-DR expression in PB-Mon from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI). To quantify the proinflammatory (CD14+loCD16+) and classical (CD14+hiCD16-) PB-Mon subsets, and to search for abnormal CD56 expression on PB-Mon from LR-MDS patients.
Methods
Fourteen patients with LR-MDS (8 males, median age 76 years), and an equal number of NI (blood donors, 8 males, median age 55 years) were studied. Patients who were being treated with myeloid growth factors were excluded, as did patients with active infections and other concomitant neoplasms. The median time from the diagnosis was 7.6 years, ranging from 0.5 to 12.6 years. Seven patients had refractory anemia with ringed sideroblasts (RARS), 4 patients had refractory cytopenias with unilineage dysplasia (RCUD), and 3 patients had refractory cytopenias with multilineage dysplasia (RCMD). Eight patients had low IPSS risk and 6 patients had intermediate 1 IPSS risk. The median PB-Mon count was of 426/mm3 (17 to 1132), in LR-MDS (>1000/mm3 in 1 case) and 482/mm3 (294 to 937), in NI (no cases with >1000/mm3).PB samples were collected into EDTA-K3 containing tubes. Cell immunophenotyping was performed by 8-color FC using fluorochrome conjugated monoclonal antibodies with different specificities (T1: CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-APC, CD14-APC-H7, CD16-V450, CD45-KO; T2: HLA-DR-FITC, CD64-PE, CD34-PerCPCy5.5, CD56-PC7, CD11c-APC, CD14-APC-H7, CD16-V450, CD45-KO), and a whole blood stain-lyse-and-then wash method (FACSLysing, Becton Dickinson–BD). A normal PB sample was run in parallel with each patient. Sample acquisition was performed in a FACSCanto II flow cytometer (BD), calibrated according to the Euroflow SOP. Data analysis was done with Infinicyt (Cytognos). Results are expressed as median, minimum and maximum values of the median fluorescence intensity observed for each marker. P values < 0.05 were considered statistically significant (Mann-Whitney U test).
Results
PB-Mon from patients with LR-MDS had FSC and SSC similar to the PB-Mon from NI (p>0.05). In the same way, the overall levels of CD13, CD14, CD15, CD45 and CD64 expression on PB-Mon from patients with LR-MDS did not differ significantly from those observed in NI (p>0.05). In contrast, PB-Mon from LR-MDS patients had significantly higher levels of CD56 (p=0.006), and lower levels of CD11c (p=0.004), CD16 (p=0.005) and HLA-DR (p=0.042), and showed a tendency for a lower CD11b expression (p=0.089), as compared to NI. Additionally, PB-Mon from LR-MDS patients had a marked decrease in the fraction of CD14+loCD16+ subset (3.1% ranging from 0.1 to 8.1% and 11.5% ranging from 4.1 to 27.7%, respectively; p<0.001). Moreover, CD14+hiCD16- PB-Mon from LR-MDS patients had a significantly higher % of CD56+ cells, as compared to NI (14.8% ranging from 0.1 to 98.7% and 7.1% ranging from 0.1 to 14.9%, respectively; p=0.026).
Conclusion
PB-Mon from patients with LR-MDS have abnormal levels of CD11b, CD11c, CD16 and CD56 expression and a high classical / proinflammatory Mon subset ratio.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Monocyte
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are hematologic neoplasms characterized by morphologic dysplasia and peripheral blood (PB) cytopenias. Flow cytometry (FC) studies have revealed that bone marrow monocytes (Mon) from MDS patients have abnormal maturation patterns and aberrant antigen expression. However, the immunophenotypic alterations in PB monocytes (PB-Mon) have been much less explored.
Aims
To evaluate the levels of CD11b, CD11c, CD13, CD14, CD15, CD16, CD64 and HLA-DR expression in PB-Mon from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI). To quantify the proinflammatory (CD14+loCD16+) and classical (CD14+hiCD16-) PB-Mon subsets, and to search for abnormal CD56 expression on PB-Mon from LR-MDS patients.
Methods
Fourteen patients with LR-MDS (8 males, median age 76 years), and an equal number of NI (blood donors, 8 males, median age 55 years) were studied. Patients who were being treated with myeloid growth factors were excluded, as did patients with active infections and other concomitant neoplasms. The median time from the diagnosis was 7.6 years, ranging from 0.5 to 12.6 years. Seven patients had refractory anemia with ringed sideroblasts (RARS), 4 patients had refractory cytopenias with unilineage dysplasia (RCUD), and 3 patients had refractory cytopenias with multilineage dysplasia (RCMD). Eight patients had low IPSS risk and 6 patients had intermediate 1 IPSS risk. The median PB-Mon count was of 426/mm3 (17 to 1132), in LR-MDS (>1000/mm3 in 1 case) and 482/mm3 (294 to 937), in NI (no cases with >1000/mm3).PB samples were collected into EDTA-K3 containing tubes. Cell immunophenotyping was performed by 8-color FC using fluorochrome conjugated monoclonal antibodies with different specificities (T1: CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-APC, CD14-APC-H7, CD16-V450, CD45-KO; T2: HLA-DR-FITC, CD64-PE, CD34-PerCPCy5.5, CD56-PC7, CD11c-APC, CD14-APC-H7, CD16-V450, CD45-KO), and a whole blood stain-lyse-and-then wash method (FACSLysing, Becton Dickinson–BD). A normal PB sample was run in parallel with each patient. Sample acquisition was performed in a FACSCanto II flow cytometer (BD), calibrated according to the Euroflow SOP. Data analysis was done with Infinicyt (Cytognos). Results are expressed as median, minimum and maximum values of the median fluorescence intensity observed for each marker. P values < 0.05 were considered statistically significant (Mann-Whitney U test).
Results
PB-Mon from patients with LR-MDS had FSC and SSC similar to the PB-Mon from NI (p>0.05). In the same way, the overall levels of CD13, CD14, CD15, CD45 and CD64 expression on PB-Mon from patients with LR-MDS did not differ significantly from those observed in NI (p>0.05). In contrast, PB-Mon from LR-MDS patients had significantly higher levels of CD56 (p=0.006), and lower levels of CD11c (p=0.004), CD16 (p=0.005) and HLA-DR (p=0.042), and showed a tendency for a lower CD11b expression (p=0.089), as compared to NI. Additionally, PB-Mon from LR-MDS patients had a marked decrease in the fraction of CD14+loCD16+ subset (3.1% ranging from 0.1 to 8.1% and 11.5% ranging from 4.1 to 27.7%, respectively; p<0.001). Moreover, CD14+hiCD16- PB-Mon from LR-MDS patients had a significantly higher % of CD56+ cells, as compared to NI (14.8% ranging from 0.1 to 98.7% and 7.1% ranging from 0.1 to 14.9%, respectively; p=0.026).
Conclusion
PB-Mon from patients with LR-MDS have abnormal levels of CD11b, CD11c, CD16 and CD56 expression and a high classical / proinflammatory Mon subset ratio.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Monocyte
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