LOW LEVELS OF CD10, CD11B, CD13, AND CD16 EXPRESSION IN THE PERIPHERAL BLOOD NEUTROPHILS FROM PATIENTS WITH LOWER RISK MYELODYSPLASTIC SYNDROMES
(Abstract release date: 05/19/16)
EHA Library. Cabral R. 06/09/16; 134813; PB1913
Prof. Renata Cabral
Contributions
Contributions
Abstract
Abstract: PB1913
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic neoplasms characterized by morphologic dysplasia, aberrant hematopoiesis and peripheral blood (PB) cytopenias, and an increased probability of transformation to acute leukemia. Diagnosis of MDS relies on well-defined cytological and cytogenetic criteria but is challenging in a significant number of patients. The detection of abnormal maturation patterns and aberrant antigen expression in the bone marrow (BM) cells has been extensively studied by flow cytometry (FC) and is now considered a promising tool to improve MDS diagnostics. However, the value of immunophenotyping the PB cells from patients with MDS has been largely ignored. Having regard to accessibility of PB samples, it would be useful to establish FC criteria for the diagnosis of MDS in the PB.
Aims
To evaluate the levels of CD10, CD11b, CD13, CD15, CD16 and CD45 expression in PB neutrophils (PB-Neut) from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI).
Methods
Fourteen patients with previously diagnosed 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-Neut count was of 2590/mm3 (365 to 6945), in LR-MDS (<1500/mm3 in 4 cases) and 4181/mm3 (3083 to 8633), in NI (no cases with <1500/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 (CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-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 PB sample. 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-Neut from patients with LR-MDS had significantly lower FSC (p=0.008) and SSC (p<0.001), as compared to those of NI. In addition, the levels of CD10 and CD11b (p<0.001 in both cases), CD16 (p=0.002) and CD13 (p=0.022) expression in PB-Neut from patients with LR-MDS were significantly decreased in patients with LR-MDS, as compared with NI. No significant differences were observed for CD15 and CD45 expression (p>0.05).
Conclusion
PB-Neut immunophenotyping may provide useful information for the diagnosis of MDS, as a complement to cytomorphology. Each center should establish its own normal reference values, on the basis of monoclonal antibodies (clones, fluorochromes) and experimental conditions used. In addition, should be given special attention to the conditions of calibration and stability of the cytometer.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Neutrophil
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic neoplasms characterized by morphologic dysplasia, aberrant hematopoiesis and peripheral blood (PB) cytopenias, and an increased probability of transformation to acute leukemia. Diagnosis of MDS relies on well-defined cytological and cytogenetic criteria but is challenging in a significant number of patients. The detection of abnormal maturation patterns and aberrant antigen expression in the bone marrow (BM) cells has been extensively studied by flow cytometry (FC) and is now considered a promising tool to improve MDS diagnostics. However, the value of immunophenotyping the PB cells from patients with MDS has been largely ignored. Having regard to accessibility of PB samples, it would be useful to establish FC criteria for the diagnosis of MDS in the PB.
Aims
To evaluate the levels of CD10, CD11b, CD13, CD15, CD16 and CD45 expression in PB neutrophils (PB-Neut) from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI).
Methods
Fourteen patients with previously diagnosed 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-Neut count was of 2590/mm3 (365 to 6945), in LR-MDS (<1500/mm3 in 4 cases) and 4181/mm3 (3083 to 8633), in NI (no cases with <1500/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 (CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-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 PB sample. 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-Neut from patients with LR-MDS had significantly lower FSC (p=0.008) and SSC (p<0.001), as compared to those of NI. In addition, the levels of CD10 and CD11b (p<0.001 in both cases), CD16 (p=0.002) and CD13 (p=0.022) expression in PB-Neut from patients with LR-MDS were significantly decreased in patients with LR-MDS, as compared with NI. No significant differences were observed for CD15 and CD45 expression (p>0.05).
Conclusion
PB-Neut immunophenotyping may provide useful information for the diagnosis of MDS, as a complement to cytomorphology. Each center should establish its own normal reference values, on the basis of monoclonal antibodies (clones, fluorochromes) and experimental conditions used. In addition, should be given special attention to the conditions of calibration and stability of the cytometer.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Neutrophil
Abstract: PB1913
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic neoplasms characterized by morphologic dysplasia, aberrant hematopoiesis and peripheral blood (PB) cytopenias, and an increased probability of transformation to acute leukemia. Diagnosis of MDS relies on well-defined cytological and cytogenetic criteria but is challenging in a significant number of patients. The detection of abnormal maturation patterns and aberrant antigen expression in the bone marrow (BM) cells has been extensively studied by flow cytometry (FC) and is now considered a promising tool to improve MDS diagnostics. However, the value of immunophenotyping the PB cells from patients with MDS has been largely ignored. Having regard to accessibility of PB samples, it would be useful to establish FC criteria for the diagnosis of MDS in the PB.
Aims
To evaluate the levels of CD10, CD11b, CD13, CD15, CD16 and CD45 expression in PB neutrophils (PB-Neut) from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI).
Methods
Fourteen patients with previously diagnosed 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-Neut count was of 2590/mm3 (365 to 6945), in LR-MDS (<1500/mm3 in 4 cases) and 4181/mm3 (3083 to 8633), in NI (no cases with <1500/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 (CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-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 PB sample. 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-Neut from patients with LR-MDS had significantly lower FSC (p=0.008) and SSC (p<0.001), as compared to those of NI. In addition, the levels of CD10 and CD11b (p<0.001 in both cases), CD16 (p=0.002) and CD13 (p=0.022) expression in PB-Neut from patients with LR-MDS were significantly decreased in patients with LR-MDS, as compared with NI. No significant differences were observed for CD15 and CD45 expression (p>0.05).
Conclusion
PB-Neut immunophenotyping may provide useful information for the diagnosis of MDS, as a complement to cytomorphology. Each center should establish its own normal reference values, on the basis of monoclonal antibodies (clones, fluorochromes) and experimental conditions used. In addition, should be given special attention to the conditions of calibration and stability of the cytometer.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Neutrophil
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic neoplasms characterized by morphologic dysplasia, aberrant hematopoiesis and peripheral blood (PB) cytopenias, and an increased probability of transformation to acute leukemia. Diagnosis of MDS relies on well-defined cytological and cytogenetic criteria but is challenging in a significant number of patients. The detection of abnormal maturation patterns and aberrant antigen expression in the bone marrow (BM) cells has been extensively studied by flow cytometry (FC) and is now considered a promising tool to improve MDS diagnostics. However, the value of immunophenotyping the PB cells from patients with MDS has been largely ignored. Having regard to accessibility of PB samples, it would be useful to establish FC criteria for the diagnosis of MDS in the PB.
Aims
To evaluate the levels of CD10, CD11b, CD13, CD15, CD16 and CD45 expression in PB neutrophils (PB-Neut) from patients with lower risk MDS (LR-MDS), as compared to normal individuals (NI).
Methods
Fourteen patients with previously diagnosed 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-Neut count was of 2590/mm3 (365 to 6945), in LR-MDS (<1500/mm3 in 4 cases) and 4181/mm3 (3083 to 8633), in NI (no cases with <1500/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 (CD15-FITC, CD13-PE, CD34-PerCPCy5.5, CD10-PC7, CD11b-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 PB sample. 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-Neut from patients with LR-MDS had significantly lower FSC (p=0.008) and SSC (p<0.001), as compared to those of NI. In addition, the levels of CD10 and CD11b (p<0.001 in both cases), CD16 (p=0.002) and CD13 (p=0.022) expression in PB-Neut from patients with LR-MDS were significantly decreased in patients with LR-MDS, as compared with NI. No significant differences were observed for CD15 and CD45 expression (p>0.05).
Conclusion
PB-Neut immunophenotyping may provide useful information for the diagnosis of MDS, as a complement to cytomorphology. Each center should establish its own normal reference values, on the basis of monoclonal antibodies (clones, fluorochromes) and experimental conditions used. In addition, should be given special attention to the conditions of calibration and stability of the cytometer.
Session topic: E-poster
Keyword(s): Flow cytometry, MDS, Neutrophil
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