CHARACTERISTICS OF BLASTS IN THE PERIPHERAL BLOOD AND BONE MARROW SAMPLES OF THE PATIENTS WITH MDS TREATED WITH 5-AZACITADINE
(Abstract release date: 05/21/15)
EHA Library. Mya H. 06/12/15; 102742; PB1834
Disclosure(s): Hospital Universitario de SalamancaHaematology
Hae Tha Mya
Contributions
Contributions
Abstract
Abstract: PB1834
Type: Publication Only
Background
Identification of the blast population in the patients with MDS has been improved in the recent years with the application of multiparametric flow cytometry (MFC).
Aims
The aim of our study was to fully characterize the blasts in the peripheral blood (PB) and bone marrow (BM) of the patients with MDS, by MFC before and after the treatment with 5-Azacitadine (5-AZA).
Methods
Patients with MDS, suitable for the treatment with 5-AZA were selected by the participating centers after the approval of the ethic committee. BM and PB samples were analyzed by BD FACSCanto flow cytometer before and after the treatment with 5-AZA. For each sample, detailed analysis with Euroflow leukemia orientation tube and acute myeloid leukaemia panels were performed (Table 1). A minimum of two million events were acquired. Infinicyt software version 1.8 (Euroflow member version) was used to analyze the acquired data files.
Results
Thirteen patients have consented to participate in the study from January 2011 to December 2014. Eight patients had normal cytogenetics and the remainder had one to three abnormal clones with del 5q, del 20q, trisomy 8 and del 11q. All patients had advanced MDS, either RAEB-1 or RAEB2 at 5-AZA treatment and received at least 3 cycles of 5-AZA before the reassessment.There were 13 pre and post 5-AZA BM samples and 8 pre and post 5-AZA PB samples. The pre 5-AZA blast count by flow cytometry ranged from 2.5 to 14% in the bone marrow and post 5-AZA blast count from 0.48% to 13%. By applying the Euroflow panels, the blast population with the same expression pattern as in the marrow was detected in the peripheral blood samples of all patients, with the lowest post treatment peripheral blood blast count being 0,02%. The blasts were uniformly positive for CD117, HLADR, dim CD45, CD13, CD33 and CD34 except one patient with negative CD34 expression. They did not express granulocytic or monocytic maturation markers in all patients, with MPO expression found only in 28% of the tested samples. Dim CD71 and dim CD123 expression was noted in ten patients whereas aberrant CD7 expression in three patients, CD56 in one patient and CD22 in one patient were noted. None of the patients had cCD3 or CD19 expression.
Summary
The myeloblasts in the study patients demonstrated more immature immunophenotypic features with rare expression of maturation markers, consistent with the previous findings in advanced MDS. MFC can accurately identify the blast population of the same immunophenotypic features as that of the BM samples, in the PB samples at diagnosis and post treatment. Assessing the evolution of the characteristic blast population in the PB samples by multiparameter flow-cytometry may be an interesting and useful approach in the patients with MDS during the post-treatment period.
Keyword(s): Flow cytometry, Myelodysplasia
Type: Publication Only
Background
Identification of the blast population in the patients with MDS has been improved in the recent years with the application of multiparametric flow cytometry (MFC).
Aims
The aim of our study was to fully characterize the blasts in the peripheral blood (PB) and bone marrow (BM) of the patients with MDS, by MFC before and after the treatment with 5-Azacitadine (5-AZA).
Methods
Patients with MDS, suitable for the treatment with 5-AZA were selected by the participating centers after the approval of the ethic committee. BM and PB samples were analyzed by BD FACSCanto flow cytometer before and after the treatment with 5-AZA. For each sample, detailed analysis with Euroflow leukemia orientation tube and acute myeloid leukaemia panels were performed (Table 1). A minimum of two million events were acquired. Infinicyt software version 1.8 (Euroflow member version) was used to analyze the acquired data files.
Results
Thirteen patients have consented to participate in the study from January 2011 to December 2014. Eight patients had normal cytogenetics and the remainder had one to three abnormal clones with del 5q, del 20q, trisomy 8 and del 11q. All patients had advanced MDS, either RAEB-1 or RAEB2 at 5-AZA treatment and received at least 3 cycles of 5-AZA before the reassessment.There were 13 pre and post 5-AZA BM samples and 8 pre and post 5-AZA PB samples. The pre 5-AZA blast count by flow cytometry ranged from 2.5 to 14% in the bone marrow and post 5-AZA blast count from 0.48% to 13%. By applying the Euroflow panels, the blast population with the same expression pattern as in the marrow was detected in the peripheral blood samples of all patients, with the lowest post treatment peripheral blood blast count being 0,02%. The blasts were uniformly positive for CD117, HLADR, dim CD45, CD13, CD33 and CD34 except one patient with negative CD34 expression. They did not express granulocytic or monocytic maturation markers in all patients, with MPO expression found only in 28% of the tested samples. Dim CD71 and dim CD123 expression was noted in ten patients whereas aberrant CD7 expression in three patients, CD56 in one patient and CD22 in one patient were noted. None of the patients had cCD3 or CD19 expression.
Table 1. Monoclonal antibodies used to analyze the blasts of in bone marrow and peripheral blood samples
Tubes | FITC | PE | PerCP5.5 | PECy7 | APC | APCH7 | PB | PO |
Tube 1 | CD16 | CD13 | CD34 | CD117 | CD11b | CD10 | HLA-DR | CD45 |
Tube 2 | CD35 | CD64 | CD34 | CD117 | IREM-2 | CD14 | HLA-DR | CD45 |
Tube 3 | CD36 | CD105 | CD34 | CD117 | CD71 | CD33 | HLA-DR | CD45 |
Tube 4 | cTDT | CD56 | CD34 | CD117 | CD7 | CD19 | HLA-DR | CD45 |
Tube 5 | CD15 | 7.1 | CD34 | CD117 | CD22 | CD38 | HLA-DR | CD45 |
Tube 6 | CD42b/CD61 | CD203c | CD34 | CD117 | CD123 | CD4 | HLA-DR | CD45 |
Tube 7 | cMPO | cCD79b | CD34 | CD19 | CD7 | cCD3 | HLA-DR | CD45 |
Summary
The myeloblasts in the study patients demonstrated more immature immunophenotypic features with rare expression of maturation markers, consistent with the previous findings in advanced MDS. MFC can accurately identify the blast population of the same immunophenotypic features as that of the BM samples, in the PB samples at diagnosis and post treatment. Assessing the evolution of the characteristic blast population in the PB samples by multiparameter flow-cytometry may be an interesting and useful approach in the patients with MDS during the post-treatment period.
Keyword(s): Flow cytometry, Myelodysplasia
Abstract: PB1834
Type: Publication Only
Background
Identification of the blast population in the patients with MDS has been improved in the recent years with the application of multiparametric flow cytometry (MFC).
Aims
The aim of our study was to fully characterize the blasts in the peripheral blood (PB) and bone marrow (BM) of the patients with MDS, by MFC before and after the treatment with 5-Azacitadine (5-AZA).
Methods
Patients with MDS, suitable for the treatment with 5-AZA were selected by the participating centers after the approval of the ethic committee. BM and PB samples were analyzed by BD FACSCanto flow cytometer before and after the treatment with 5-AZA. For each sample, detailed analysis with Euroflow leukemia orientation tube and acute myeloid leukaemia panels were performed (Table 1). A minimum of two million events were acquired. Infinicyt software version 1.8 (Euroflow member version) was used to analyze the acquired data files.
Results
Thirteen patients have consented to participate in the study from January 2011 to December 2014. Eight patients had normal cytogenetics and the remainder had one to three abnormal clones with del 5q, del 20q, trisomy 8 and del 11q. All patients had advanced MDS, either RAEB-1 or RAEB2 at 5-AZA treatment and received at least 3 cycles of 5-AZA before the reassessment.There were 13 pre and post 5-AZA BM samples and 8 pre and post 5-AZA PB samples. The pre 5-AZA blast count by flow cytometry ranged from 2.5 to 14% in the bone marrow and post 5-AZA blast count from 0.48% to 13%. By applying the Euroflow panels, the blast population with the same expression pattern as in the marrow was detected in the peripheral blood samples of all patients, with the lowest post treatment peripheral blood blast count being 0,02%. The blasts were uniformly positive for CD117, HLADR, dim CD45, CD13, CD33 and CD34 except one patient with negative CD34 expression. They did not express granulocytic or monocytic maturation markers in all patients, with MPO expression found only in 28% of the tested samples. Dim CD71 and dim CD123 expression was noted in ten patients whereas aberrant CD7 expression in three patients, CD56 in one patient and CD22 in one patient were noted. None of the patients had cCD3 or CD19 expression.
Summary
The myeloblasts in the study patients demonstrated more immature immunophenotypic features with rare expression of maturation markers, consistent with the previous findings in advanced MDS. MFC can accurately identify the blast population of the same immunophenotypic features as that of the BM samples, in the PB samples at diagnosis and post treatment. Assessing the evolution of the characteristic blast population in the PB samples by multiparameter flow-cytometry may be an interesting and useful approach in the patients with MDS during the post-treatment period.
Keyword(s): Flow cytometry, Myelodysplasia
Type: Publication Only
Background
Identification of the blast population in the patients with MDS has been improved in the recent years with the application of multiparametric flow cytometry (MFC).
Aims
The aim of our study was to fully characterize the blasts in the peripheral blood (PB) and bone marrow (BM) of the patients with MDS, by MFC before and after the treatment with 5-Azacitadine (5-AZA).
Methods
Patients with MDS, suitable for the treatment with 5-AZA were selected by the participating centers after the approval of the ethic committee. BM and PB samples were analyzed by BD FACSCanto flow cytometer before and after the treatment with 5-AZA. For each sample, detailed analysis with Euroflow leukemia orientation tube and acute myeloid leukaemia panels were performed (Table 1). A minimum of two million events were acquired. Infinicyt software version 1.8 (Euroflow member version) was used to analyze the acquired data files.
Results
Thirteen patients have consented to participate in the study from January 2011 to December 2014. Eight patients had normal cytogenetics and the remainder had one to three abnormal clones with del 5q, del 20q, trisomy 8 and del 11q. All patients had advanced MDS, either RAEB-1 or RAEB2 at 5-AZA treatment and received at least 3 cycles of 5-AZA before the reassessment.There were 13 pre and post 5-AZA BM samples and 8 pre and post 5-AZA PB samples. The pre 5-AZA blast count by flow cytometry ranged from 2.5 to 14% in the bone marrow and post 5-AZA blast count from 0.48% to 13%. By applying the Euroflow panels, the blast population with the same expression pattern as in the marrow was detected in the peripheral blood samples of all patients, with the lowest post treatment peripheral blood blast count being 0,02%. The blasts were uniformly positive for CD117, HLADR, dim CD45, CD13, CD33 and CD34 except one patient with negative CD34 expression. They did not express granulocytic or monocytic maturation markers in all patients, with MPO expression found only in 28% of the tested samples. Dim CD71 and dim CD123 expression was noted in ten patients whereas aberrant CD7 expression in three patients, CD56 in one patient and CD22 in one patient were noted. None of the patients had cCD3 or CD19 expression.
Table 1. Monoclonal antibodies used to analyze the blasts of in bone marrow and peripheral blood samples
Tubes | FITC | PE | PerCP5.5 | PECy7 | APC | APCH7 | PB | PO |
Tube 1 | CD16 | CD13 | CD34 | CD117 | CD11b | CD10 | HLA-DR | CD45 |
Tube 2 | CD35 | CD64 | CD34 | CD117 | IREM-2 | CD14 | HLA-DR | CD45 |
Tube 3 | CD36 | CD105 | CD34 | CD117 | CD71 | CD33 | HLA-DR | CD45 |
Tube 4 | cTDT | CD56 | CD34 | CD117 | CD7 | CD19 | HLA-DR | CD45 |
Tube 5 | CD15 | 7.1 | CD34 | CD117 | CD22 | CD38 | HLA-DR | CD45 |
Tube 6 | CD42b/CD61 | CD203c | CD34 | CD117 | CD123 | CD4 | HLA-DR | CD45 |
Tube 7 | cMPO | cCD79b | CD34 | CD19 | CD7 | cCD3 | HLA-DR | CD45 |
Summary
The myeloblasts in the study patients demonstrated more immature immunophenotypic features with rare expression of maturation markers, consistent with the previous findings in advanced MDS. MFC can accurately identify the blast population of the same immunophenotypic features as that of the BM samples, in the PB samples at diagnosis and post treatment. Assessing the evolution of the characteristic blast population in the PB samples by multiparameter flow-cytometry may be an interesting and useful approach in the patients with MDS during the post-treatment period.
Keyword(s): Flow cytometry, Myelodysplasia
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