Predictors for prognosis of chronic myelocytic leukemia.
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Bone marrows from 30 newly diagnosed Ph+chronic myelocytic leukemia (CML) (21 in chronic phase, CML-CP, 9 in accelerate phase, CML-AP) and 8 followed up patients in blast crisis (CML-BC) were tested by DNA strand breaks (%D value), DNA-aneuploidy, flow cytometry-cell surface antigen expression for CD15 and HLA-DR ratio. Our results showed that all these three parameters changed as the disease escalated. A very low value of %D and DNA-aneuploidy occurrences were high risk factors. Cell surface antigen expression CD15 and HLA-DR ratio measurement was simple and reliable. The ratio < 1.0 appeared earlier than morphology clarifying CML-AP and should be followed up regularly.Keywords:
CD15
Myelocytic leukemia
Blast Crisis
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CD15
Cluster of differentiation
Immunofluorescence
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Bone marrows from 30 newly diagnosed Ph+chronic myelocytic leukemia (CML) (21 in chronic phase, CML-CP, 9 in accelerate phase, CML-AP) and 8 followed up patients in blast crisis (CML-BC) were tested by DNA strand breaks (%D value), DNA-aneuploidy, flow cytometry-cell surface antigen expression for CD15 and HLA-DR ratio. Our results showed that all these three parameters changed as the disease escalated. A very low value of %D and DNA-aneuploidy occurrences were high risk factors. Cell surface antigen expression CD15 and HLA-DR ratio measurement was simple and reliable. The ratio < 1.0 appeared earlier than morphology clarifying CML-AP and should be followed up regularly.
CD15
Myelocytic leukemia
Blast Crisis
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Objective To detect the cell membrane and cell plasma antigens by flow cytometry and(evaluate) the role of these antigens in acute leukemia(AL) immunophenotyping.Methods The bone marrow or blood in 290 cases of patients with AL were tested by three color direct immunofluorescence staining to determine the immunophenotype by flow cytometry. Results The positive expression rate of CD_(13) and CD_(33) found in acute myelogenous leukemia(AML)-M_1,AML-M_2,AML-M_3,AML-M_5 was 89.6% and 77.6% and myeloperoxidase(MPO) expressions were 99.25% in 134 cases with AML.In 125 B-ALL patients, the positive rate in CyCD_(79a) was 97.6%,CD_(19) 84%,CD_(22) 84%,CD_(10) 68.8%.In 31 T-ALL patients,the positive rate in CyCD_3 was 96.8%;CD_2 71%;CD_3 80.6%;CD_5 87.1%;CD_7 83.9%.The positive expression rate of CD_(34) was 47% and 62.8% in 134 cases with AML and 156 cases with ALL.Conclusion Detection of membrane antigens and cytoplasmic antigens by flow cytometry will benefit the classification of acute leukemia.
Immunophenotyping
Immunofluorescence
Cytometry
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Myelocytic leukemia
Blast Crisis
Acute myeloblastic leukemia
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It is of common knowledge that most leukemic patients succumb to infectious complications beside dysfunction in proliferation/differentiation of hematopoietic cells.Recently, it has been shown by several groups that there is a locus specific HLA class I downregulation in the leukemic cells.However, the HLA status of the phenotypically/morphologically normal granulocytes which can cope up with the infectious complications are not known.Therefore it may be worthwhile to study the HLA status in these cell types.We, therefore, investigated the status of HLA-ABC and HLA-DR in the CD15 + granulocytes and observed a higher expression of HLA-DR in several leukemic samples in comparison to normal volunteers (NV).Our data also suggest that only CD15 + granulocytes of myeloid leukemia, a clonal stem cell disorder, have a tendency of decreased HLA class Ia antigen expression.Moreover CD15 + granulocytes of NV showed an enhanced HLA-DR expression in presence of leukemic cells.Interestingly, CD15 + granulocytes collected from normal volunteers were observed to have phagocytic oxidative burst activity towards HLA class Ia downregulated primary leukemic cells.We therefore suggest that neutrophil transplantation may be used for the treatment of leukemia.
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HLA-DR
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Cluster of differentiation
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Abstract The aim of this study was to investigate to which extent acute leukemias could be monitored for residual disease by using atypical antigen combinations as leukemia‐related markers. Atypical antigenic features were determined by double color flow cytometry and included coexpression of lymphoid and myeloid related antigens, unnphysiological coexpression of immature and mature antigens, and lack of an antigen that is normally expressed during maturation. Atypical immunophenotypes were detected in 35 of 68 patients with AML (51.5%) and 15 of 24 patients with ALL (62.5%). When 12 patients with leukemia‐associated markers were again analyzed at relapse, the relevant antigen combinations were retained in 11 of them. The sensitivity of this two color flow cytometric assay as determined in dilution experiments was 1 in 10 3 to 10 4 cells Follow‐up studies of bone marrow samples revealed that, after induction chemotherapy cells with leukemia‐associated markers were detectable in several patients at a frequency of 0.5 to 4%, but only patients in whom the cells with atypical antigens never disappeared suffered from relapse. In contrast, patients who became negative for the atypical cells remained in complete remission (median remission duration after the first negative bone marrow assessment by flow cytometry 52 weeks, range 20–102) We conclude that atypical antigen combinations, which are present in a meaningful number of acute leukemias, are a valuable means of monitoring acute leukemia patients during follow‐up. This flow cytometric approach can complement other strategies to get a more accurate definition of remission in acute leukemia. © 1992 Wiley‐Liss, Inc.
Minimal Residual Disease
Immunophenotyping
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Bone marrow cellular DNA content was measured by flow cytometry in 107 adult patients with various types of acute leukemia. DNA-aneuploidy was detected in 28 of 107 patients (26%). In 18 patients with DNA-aneuploidy bone marrow was examined throughout 2-36 months. DNA-flow cytometry of bone marrow allows to determine DNA content in leukemia cells and distinguish hypodiploid, diploid, hyperdiploid and tetraploid DNA clones in patients with acute leukemia. In some cases this method allows monitoring of the minimal residual disease.
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The immunologic phenotypes of 68 cases of acute nonlymphoblastic leukemia (ANLL) were analysed with a panel of monoclonal antibodies. The results showed that the frequencies of myeloid antigens expressed on ANLL cells are CD15 > CD33 > CD13 > CD14 and there were few cases expressing lymphoid antigens. The expression of myeloid antigen on leukemia cells is not only the evidence for diagnosis ANLL, but also is valuable predicting prognosis. In cases with the ratio of CD33/CD13 < 1, the complete remission (CR) rate was higher than those with CD33/CD13 > 1 (P < 0.05). Patients with expression of CD15 antigen on their leukemia cells appeared to be associated with long survival.
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The presence of alloantibodies against human leucocyte antigen (HLA)‐I and HLA‐II antigens has been associated with hyperacute and accelerated graft rejections. However, occasionally these rejections occur in patients without donor‐specific anti‐HLA antibodies, suggesting the presence of other antigenic complexes that are shared by the graft and other cell populations. Usually, these antibodies are not routinely studied and their role in graft rejection is poorly understood. For this reason, we tested, by flow cytometry, the presence of panel‐reactive alloantibodies (PRA) using different cell populations in 30 pre‐transplant sera of kidney graft recipients. The patients studied had or had not hyperacute and accelerated rejection episodes (HARE) and did not have alloantibodies against HLA of their specific donors. We found that IgG and IgM alloantibodies directed against HLA‐I antigens, different to the HLA‐I antigens of the specific donors, as well as IgG against endothelium/monocyte antigens, IgM against platelets, and IgM against T cells are significantly associated with HARE, independently of the percentage of PRA. Our findings suggest that the detection of antibodies by flow cytometry against non‐major histocompatibilty complex antigens may be useful as a pre‐transplant crossmatch in living related donor kidney transplants to diminish the incidence of HARE.
Panel reactive antibody
Isoantibodies
Histocompatibility
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