Immunophenotypic characteristics of acute leukaemia after myelodysplastic syndromes.
Margarida LimaTeixeira Mdos ASara MoráisMarina Gabriela Monteiro Carvalho Mori da CunhaJ. O. CoutinhoLeandro Kasuki Jomori de PinhoPatrícia RibeiroB. Justiça
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To analyse the immunophenotype of acute leukaemia (AL) after myelodysplastic syndromes (MDS) (MDS-AL) and to compare the immunophenotypic profile of acute myeloblastic leukaemia (AML) secondary to MDS (MDS-AML) with that of "de novo"-AML.Twenty patients with MDS-AL and 29 patients with "de novo"-AML were studied. Morphocytochemical and flow cytometric studies were done in each case.All the MDS-AL studied displayed a myeloid phenotype (MDS-AML). The main difference between MDS-AML and "de novo"-AML was a significantly higher frequency of CD34 expression in the first group. Differences concerning the expression of other non-lineage related or myeloid-associated markers were not statistically significant, although the percentage of cases CD15(+) was lower in MDS-AML. The overall frequency of expression of lymphoid-associated markers was similar in both groups, T-cell markers being more frequently detected.Our findings support the usefulness of immunophenotyping studies to characterize MDS-AL and suggest some immunophenotyping differences between MDS-AML and "de novo"-AML which might have biological and prognostic significance.Keywords:
Immunophenotyping
CD15
Acute myeloblastic leukemia
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To assess if the in vitro behaviour of leukaemic colony-forming units (CFU-L) from secondary leukaemias is similar to that of the de novo acute myeloblastic leukaemia. An attempt was also made to verify if such behaviour correlated with the characteristics of the disease or with the cell-surface markers of the leukaemic population.The study was carried out on 21 patients with secondary acute leukaemia (12 had previous myelodysplastic syndrome, MDS-AL, and 9 had previous chronic myeloproliferative syndromes, MPS-AL). Peripheral blood mononucleated cells were cultured on methyl-cellulose with MCL-PHA stimulation. The dishes were examined after 7 days of incubation in humid 37 degrees C environment with 5% CO2. Direct or indirect immunofluorescence was used for the immunophenotypic analysis and the patients were divided in two groups: 1) immature phenotype, which include those cases expressing only precursor (CD34) or pan-myeloid (CD33/13) antigens, and (2) mature phenotype, comprising the caes with granulomonocytic (CD15, CD14), erythroid (glycophorin) or megakaryocytic (CD61) differentiation. The statistical analysis was done with the BMDP programme.Up to 95% of the secondary acute leukaemias proliferate in vitro, as opposed to 82% of the de novo ones, the difference not being significant (p = 0.14). Successful cell showing was clearly superior in the former (p = 0.02), mostly due to higher proliferation of the MPS-AL cells. Neither the clinico-biologic characteristics of the patients nor the phenotype of the blast cells correlated with the in vitro behaviour of CFU-L. Only CD19 antigen expression and nuclear TdT provided a lesser in vitro growth (p = 0.05 and p-0.06, respectively).In general terms, secondary leukaemias, especially MPS-AL, show higher in vitro growth than de novo acute myeloblastic leukaemias.
Acute myeloblastic leukemia
Clonogenic assay
Immunophenotyping
CD15
Precursor cell
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Immunophenotyping
Univariate analysis
CD15
Chromosome abnormality
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to evaluate the biological and clinical implications of immunophenotypic abnormalities of bone marrow myeloid cell compartment in patients with primary myelodysplastic syndromes (MDS).analysis of cell surface antigen profiles was performed by flow cytometric immunophenotyping on bone marrow mononuclear cell (BMMNCs) specimens from 39 adult MDS patients and 5 healthy individuals. Expression of cell surface antigen profiles was correlated with FAB subtype, cytogenetics, CFU-GM colony growth, overall survival (OS) and leukemic transformation (LT).expression levels of early differentiation and myelo-monocytic antigens (CD38, CD13, CD33, CD14 and CD15) on myeloid cell compartment of BMMCs were significantly higher in MDS patients in comparison to healthy control group, suggesting maturational left shift of bone marrow myeloid cell compartment in MDS. CD34 antigen expression was in a positive linear correlation with HLA-DR antigen expression (r=0.652; p=0.0004), and in negative correlation with the expression of CD11b and CD15 antigens (r=-0.48; p=0.014 and r=-0.564; p=0.0033, respectively). Myeloid antigen ratio (HLA-DR/CD11b) was 2.5 fold higher in patients with MDS in comparison to control group. Patients with advanced disease had significantly higher myeloid antigen ratio than patients with low risk MDS (p<0.05). The type of CFU-GM colony growth and the presence of chromosomal aberrations were unrelated to the proportion of CD34(+) cells and elevated myeloid ratio. Patients with elevated proportion of CD34(+) BMMNCs or elevated myeloid ratio had significantly shorter OS and higher LT rate in comparison to patients whose proportion of CD34(+) BMMNCs and myeloid ratio were within normal range.the presence of abnormalities in antigen expression profiles of bone marrow myeloid cell compartment has clinical implication in MDS, with particular contribution in predicting the patient outcome. Elevated proportion of CD34(+) BMMNCs and elevated myeloid ratio of bone marrow myeloid compartment are useful immunophenotypic tools for estimation of prognosis in this very heterogeneous disorder group.
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CD15
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OBJECTIVE To investigate the quantitative expression of immunophenotype of CD34+ myeloid precursor cells in myelodysplastic syndrome (MDS) patients and its correlation with clinical characteristics, and understand the effect of quantitative expression of CD7 and CD117 on the prognosis of low-risk MDS patients. METHODS Multi-parameter flow cytometry (FCM) was used to detect the proportion and mean fluorescence intensity (MFI) of each antigen of bone marrow CD34+ myeloid precursor cells in 79 MDS patients. The correlation between the expression level of each immune marker and clinical characteristics was compared. The effects of quantitative expressions of CD7 and CD117 on the overall survival rate of low-risk patients were explored. RESULTS Bone marrow blast cell proportion (P<0.01), RBC level (P<0.01), and Hb level (P<0.05) of high-risk MDS patients were higher, while EPO level (P<0.05) was lower than those of low-risk patients. The proportion of CD34+ blast cells (P<0.01), the proportion of CD117 (P<0.05) and the MFI of CD7 (P<0.05) were higher in high-risk patients than those in low-risk patients, but the MFI of CD123 was lower (P<0.05). In high-risk MDS patients, CD15/CD34 (MFI) and CD19/CD34 (MFI) positively correlated with the proportion of total T cells (r=0.458; r=0.505), while CD19/CD34 (%) and CD19/CD34 (MFI) negatively correlated with WBC levels (r=-0.469; r=-0.503). In low-risk MDS patients, CD34+ (%) positively correlated with bone marrow erythrocyte proportion, PLT level and neutrophil level (r=0.426; r=0.486; r=0.495), but negatively correlated with LDH level (r=-0.421); WT1 expression level was positively correlated with CD10/CD34 (%), CD10/CD34 (MFI) and CD117/CD34 (MFI) (r=0.745; r=0.800; r=0.434), while negatively correlated with CD11b/CD34 (%)(r=-0.457); CD19/CD34 (%) and CD71/CD34 (MFI) negatively correlated with NK cell proportion (r=-0.786; r=-0.514); CD10/CD34 (%) positively correlated with Th/Ts, while CD7/CD34 (MFI) negatively correlated with the proportion of Th cells (r=0.738; r=-0.513); HLADR/CD34 (%) and HLADR/CD34 (MFI) negatively correlated with PLT level (r=-0.461; r=-0.445), while HLADR/CD34 (MFI) positively correlated with bone marrow NAP fraction (r=0.552). The quantitative expression of CD7 and CD117 had no significant effect on the overall survival rate of low-risk MDS patients. CONCLUSION The immunophenotype of CD34+ myeloid precursor cell in different risk groups in MDS patients is related to clinical characteristics. Bone marrow cell morphology, clinical and laboratory features and immunophenotype will be of great significance to the diagnosis, clinical classification and prognosis evaluation of MDS patients.
Immunophenotyping
CD117
Precursor cell
Interleukin-3 receptor
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The objective of this study was to investigate the biological characteristics and the therapeutic effect in patients with acute erythroleukemia (AML-M(6)). Morphology, immunophenotype and cytogenetics were retrospectively analyzed in 29 patients with AML-M(6) and were compared with 30 AML-M(2) patients. The results showed that there were immature cells (2% - 10%) and erythroblast, and puncture of bone marrow revealed myelodysplastic features involving multiple hemopoietic lineages in bone marrow of 19 patients. Flow cytometry indicated that the expression frequency of Gly-A in M(6) significantly increased (66.67 +/- 23.86)% and higher than that in M(1), M(2), M(3), M(4) and M(5) (P < 0.01). The expression frequencies of HLA-DR (60.00 +/- 24.79%), CD34 (40.00 +/- 24.79%), CD38 (33.33 +/- 23.86%) in M(6) were high, and the frequencies of myeloid immunophenotypes CD13 (66.67 +/- 23.86%), MPO (33.33 +/- 23.86%), CD33 (46.67 +/- 25.25%), CD15 (33.33 +/- 23.86%), CD117 (46.67 +/- 25.25%) were common as well in M(6). Lymphocytic immunophenotypes CD3, CD4, CD19 were detected in part of patients with M(6), and the expression frequencies of CD4 was 26.67%. The expression frequences of CD38, CD33, CD15, MPO in M(6) were less common than that in M(2) (P < 0.01). In 4 out of 9 M(6) patients the chromosomal abnormatility (44.44%) was seen, in one of which complex chromosome abnormality was found. The complete remmision rate of M(6) patients was 29.41%, and lower than that of M(2) patients (68.18%, P < 0.01). It is concluded that Gly-A is a specific immunophenotype in M(6), which can help to distinguish M(6) from other types of acute myeloid leukemia. Poor clinical therapeutic response may correlated with its biological characteristics.
Immunophenotyping
CD15
CD117
Chromosome abnormality
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Immunophenotyping
CD15
CD117
Acute myeloblastic leukemia
Minimal Residual Disease
Precursor cell
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The phenotypes of the bone marrow cells in various subtypes of myelodysplastic syndromes (MDS) and its clinical implication were explored. The antigen expression of a panel of antigens expressed in marrow cells from 30 patients with subtypes of MDS was assayed by alkaline phosphatase anti-alkaline phosphatase method. The results showed that the expression of myeloid antigens appeared abnormality, CD13 and CD33, found on granulocyte and macrophage precursors, increased, and CD15 decreased. There were no significant changes for monocytic antigen CD14 and lymphoid antigens CD7 and CD10. CD34 was increased in RAEB/RAEB-t and was not increased in RA/RAS patients. CD71, expressed by erythroblast and proliferative cells, was higher in all subtypes of MDS than that in control group. It is suggested that the bone marrow cells from MDS patients showed abnormality of more than two series of immunophenotypes, detection of immunophenotype in MDS cells might be contributed to the diagnosis and predicting prognosis.
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CD15
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To explore the immunophenotype of myeloid granulocytes in myelodsplastic syndromes(MDS)and aplastic anemia(AA),we detected CDl3、CD33、CD15、CD10、CD34 and HLA-DR on the membrane surface of myeloid granulocytes from patients with myelodysplastic syndromes,patients with aplastic anemia and normal subjects by flow cytometry.There was an immunophenotypic misexpression of myeloid granulocytes in MDS patients.So the immunophenotype analysis of myeloid granulocytes might be useful for the diagnosis of MDS patients and AA patients.
Immunophenotyping
Aplastic anemia
CD15
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To analyze hematopoietic cell surface antigen reactivity in acute leukemia (AL) by flow cytometry and identify acute mixed-lineage leukemias (AMLL) employing the most widely accepted criteria.Ninety seven patients with de novo AL were studied. Cell surface antigens were investigated with monoclonal antibodies directed to: B lymphoid (CD10, CD19, CD20, CD21, CD22); T lymphoid (CD2, CD3, CD5, CD7); and myeloid (CD13, CD14, CD15, CD33, CD41) cell lineages. Maturation cell-associated antigens (CD34, HLA-DR and TdT) were also studied.Twelve patients unclassified by cytomorphology could be classified by immunophenotype. Using cytomorphologic, cytochemical and immunophenotypic data, 54 cases corresponded to acute lymphoblastic leukemia (ALL) and 43 were acute myeloblastic leukemia (AML). In All there were 63% B lineage, 15% T, 7% T/B, 6% undifferentiated and 9% mixed-lineage (coexpression of two or more myeloid-associated antigens). In AML, myeloid immunophenotype was observed in 86% undifferentiated in 2%, and mixed-lineage in 12% (coexpression of two or more lymphoid-associated antigens). In addition, 26% of ALL cases and 12% of AML cases expressed a single myeloid and lymphoid antigen respectively. The most common aberrant antigens in ALL and AML were CD13 and CD7 respectively. The highest frequency of CD34 antigen expression (90%) was detected in patients with AMLL.Flow cytometric immunophenotypic analysis allowed to: a) establish diagnosis in cytomorphologically unclassified cases; b) identify AMLL with a frequency similar to that reported in other series; and c) confirm the heterogeneity of AL.
Immunophenotyping
CD5
Acute myeloblastic leukemia
CD15
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To analyse the immunophenotype of acute leukaemia (AL) after myelodysplastic syndromes (MDS) (MDS-AL) and to compare the immunophenotypic profile of acute myeloblastic leukaemia (AML) secondary to MDS (MDS-AML) with that of "de novo"-AML.Twenty patients with MDS-AL and 29 patients with "de novo"-AML were studied. Morphocytochemical and flow cytometric studies were done in each case.All the MDS-AL studied displayed a myeloid phenotype (MDS-AML). The main difference between MDS-AML and "de novo"-AML was a significantly higher frequency of CD34 expression in the first group. Differences concerning the expression of other non-lineage related or myeloid-associated markers were not statistically significant, although the percentage of cases CD15(+) was lower in MDS-AML. The overall frequency of expression of lymphoid-associated markers was similar in both groups, T-cell markers being more frequently detected.Our findings support the usefulness of immunophenotyping studies to characterize MDS-AL and suggest some immunophenotyping differences between MDS-AML and "de novo"-AML which might have biological and prognostic significance.
Immunophenotyping
CD15
Acute myeloblastic leukemia
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