CD45-negative acute leukemia in adulthood (original) (raw)
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Flow cytometry CD45 gating for immunophenotyping of acute myeloid leukemia
Leukemia, 1997
A flow cytometry method has been introduced into the routine age values within the malignant blast cell populations or investigation of whole bone marrow samples following red within the total nucleated cells present in each sample. In blood cell lysis on the basis of a primary CD45/side scatter order to give numbers of malignant cells and proportions of (SSC) gating procedure. Blast cells were first identified by cells within this cohort, a reliable practical discrimination CD45/SSC gating in 74 cases of acute myeloid leukemia (AML) between malignant blasts and normal cell types would be and the results were compared to a conventional FSC/SSC gating procedure and to MGG-staining smears. The percentages required. In this paper, as previously recommended by of blast cells in these samples as defined by the morphological Borowitz et al, 6 we suggest that such a discrimination can be analysis of MGG smears correlated better with the values readily facilitated by introducing primary gating for CD45 determined by CD45/SSC gating (r ؍ 0.94) than with the blast antigen expression and side scatter (CD45/SSC). We also sugcell counts recorded with FSC/SSC gating (r ؍ 0.76). These gest that this step should replace the first gating step for forfindings were not surprising because while CD45 expression ward scatter and side scatter (FSC/SSC), as this latter procedure was regularly lower on leukemic blasts than on normal lymphoid and monocytic cells, the FCS/SSC characteristics of these does not discriminate well between leukemic blasts, lymphopopulations were overlapping. In 53 samples, the blast cell cytes and monocytes. populations were also analyzed with a panel of FITC-conju-In this paper we present the application of double and triple gated monoclonal antibodies that were utilized in double labeimmunofluorescence (IF) analysis of bone marrow samples ling with CD45-PE. We show that the CD45/SSC gating protaken from patients during the presentation of AML. With the cedure improved phenotypic determination of the blast cells in systematic use of leukocyte common antigen (CD45) marker three ways: (1) by discriminating between leukemic blast cells and residual normal cells; (2) by excluding normal cells from in combination with lineage-specific markers, a good disthe phenotypic analysis of leukemic blast cells; and (3) by crimination can be achieved between the blast cell popuidentifying blast cell heterogeneity in many cases of leukemia lations and the normal cells. This discrimination is based on on the basis of different CD45 display. Moreover, this immunothe fact that the precursor cells in the bone marrow, as well phenotyping procedure on whole bone marrow samples also as the leukemias which derive from these cell types, express allowed an efficient discrimination between the various cell linlow and intermediate values of CD45, while lymphocytes and eages and facilitated the analysis of leukemic blasts present in low proportions. monocytes express high levels of CD45. 7
Side scatter versus CD45 flow cytometric plot can distinguish acute leukaemia subtypes
Indian Journal of Medical Research, 2016
Background & objectives: Flow cytometry is an important tool to diagnose acute leukaemia. Attempts are being made to find the minimal number of antibodies for correctly diagnosing acute leukaemia subtypes. The present study was designed to evaluate the analysis of side scatter (SSC) versus CD45 flow dot plot to distinguish acute myeloid leukaemia (AML) from acute lymphoblastic leukaemia (ALL), with minimal immunological markers. Methods: One hundred consecutive cases of acute leukaemia were evaluated for blast cluster on SSC versus CD45 plots. The parameters studied included visual shape, CD45 and side scatter expression, continuity with residual granulocytes/lymphocytes/monocytes and ratio of maximum width to maximum height (w/h). The final diagnosis of ALL and AML and their subtypes was made by morphology, cytochemistry and immunophenotyping. Two sample Wilcoxon rank-sum (Mann Whitney) test and Kruskal-Wallis equality-of-populations rank tests were applied to elucidate the significance of the above ratios of blast cluster for diagnosis of ALL, AML and their subtypes. Receiver operating characteristic (ROC) curves were generated and the optimal cutoffs of the w/h ratio to distinguish between ALL and AML determined. Results: Of the 100 cases, 57 of ALL and 43 cases of AML were diagnosed. The median w/h ratio of blast population was 3.8 for ALL and 1 for AML (P<0.001). ROC had area under curve of 0.9772.The optimal cutoff of the w/h ratio for distinction of ALL from AML was found to be 1.6. Interpretation & conclusions: Our findings suggest that if w/h ratio on SSC versus CD45 plot is less than 1.6, AML may be considered, and if it is more than 1.6, ALL may be diagnosed. Using morphometric analysis of the blast cluster on SSC versus CD45, it was possible to distinguish between ALL and AML, and their subtypes.
American journal of clinical pathology, 1993
This article describes a procedure for performing routine three-color flow cytometric analysis for acute leukemia on lysed whole bone marrow preparations. This technique uses the combination of CD45 intensity and right-angle light scatter (RALS) to distinguish leukemic cells from normal lymphocytes, monocytes, neutrophils, eosinophils, and nucleated red blood cells. On this display, leukemic cells occupy a unique blast region characterized by intermediate CD45 density and low RALS, which, in normal marrows, contains less than 5% of the total cells. This approach was applied to 39 cases of acute leukemia and 8 cases of myelodysplasia or myeloproliferative disorders. The estimate of blasts by flow cytometric analysis was correlated highly with morphologic leukemic cell counts over a wide range. Moreover, the pattern seen on the CD45-RALS display was different for different French-American-British subtypes of leukemia, suggesting that this pattern might be useful for categorization. Wh...
The properties of CD 45 / SS in the blast Population of AML Sudanese patients
2015
Background: Leukemia is a group of disorders characterized by production of excessive numbers of abnormal white blood cells in the bone marrow and blood. Acute leukemia is a rapid progressive and fatal leukemia. Acute Myeloid Leukemia (AML) accounts for Sahar M. Almahal, Enaam A. Abdelgader, Osama A. Altayeb, Eman Abbass F., Amin A. Al-Amin, Rasha Abdelgleel, Gada M. A. Merghani, Tarig M. Karfis, Eldirdiri M. Abdelrhman, Osman H. Musa, Mohammed A. AbdallaThe properties of CD45/ SS in the blast Population of AML Sudanese patients EUROPEAN ACADEMIC RESEARCH Vol. III, Issue 6 / September 2015 6555 approximately 20% of acute leukemia in children and 80% of acute leukemia in adults. Immunophenotyping has become extremely important not only in diagnosis and sub classification of AML but also in the detection of the minimal residual disease. The aim of this paper was to study the role of CD45 gating strategy and its properties using flowcytometry in acute myeloid leukemia and to achieve go...
Role of blast cell immunophenotyping for the diagnosis and prognosis of acute myeloid leukemia
Biology of the Cell, 1992
Bone marrow blast cell antigen expression from 86 patients with cle n o w acute myeloid leukemias (AML) was studied and corrclated with FAB classification and clinical outcome. Among a panel of 14 monoclonal antibodies routinely used for the diagnosis of acute leukemias we studied the expression of six antibodies (CD13, CD15, VIM2, CU33, CD14, CD34) of the granulomonocytic lineage and found that some of them were useful for diagnosis and/or prognosis. For FAB subclassification of AML, the CD13 or VIM2 antigen expression was of no benefit. Monocytic leukemias (M4+ M5PD+ M5WD) more frequently expressed CD34 antigen (28/31) than granulocytic (MI +M2+M3) subtypes (33/53) (P<O.OI). Finally, the most striking differences were found with CD14 antigen expression: CD14 antigcn was more frequently expressed in M4+ M5 leukemias (21/31) than in M1 + M 2 + M3 subtypes (12/33) (P<O.OI). The mean percentage of CD14 positive blast cells was accordingly higher in monocytic leukemias than in granulocytic leukemias and the difference was highly significant (P<O.OOOI). The CD15 antigen was more frequently expressed in differentiated leukemias (M2 + M3 + M4+ M5WD) (35/44) than in poorly differentiated forms (M 1 + M5PD) (17/37) (P< 0.001). The statistical difference was higher when the mean percentage of CD15 positive blast cells were compared (P<0-0003). Moreover these latter percentages were different in M1 and M2 subtypes (P < 0.003). The blast cell expression of CDI 3, CD14, C D I5 or CD33 was not predictive of the length of CR or survival. Moreover, our results support previously published findings suggesting a longer overall survival duration for patients whose leukemic cells do not express the CD34 antigen (P<O.OI). We also confirm that patients with the more differentiated subtypes of AML (CD13-, CD34+) tend to survive longer than patients with the less differentiated subtypes ofAML (CD13--, CD34+) (P<O-OOl).
Blood, 1996
During the immunodiagnosis of 517 cases of acute myelogenous leukemia (AML) entered into the Medical Research Council (MRC) AML 10 trials, we have observed the CD34 precursor cell antigen more frequently in AML of M2 morphology, especially in the 84% of cases with the t(8;21) chromosomal translocation, than in any other French-American-British classification group. CD34 expression was then quantified (using QIFI and Quantum Simply Cellular beads [Flow Cytometry Standards, Research Triangle Park, NC] and CD34+ standard cells). When CD34 antibody-binding capacity (ABC) of normal bone marrow (BM) precursors and leukemic blasts was compared, it was shown that AML M2 cases with t(8;21) not only had the highest percentages of CD34+ blasts, but in > 80% of CD34+ cases the individual blasts expressed higher than normal levels of CD34 antigen (> 60 x 10(3) ABC per cell). In addition, in 73% of this group CD34 antigen was overexpressed in an asynchronous combination with cytoplasmic mye...
Assessment of the normal or leukemic nature of CD34+ cells in acute myeloid
2003
Background and Objectives. The percentages of CD34 + cells in the bone marrow of patients with acute myeloid leukemia (AML) vary widely. Especially in the low range (<5% CD34 + cells), the nature (normal or malignant) of the CD34 + cells is uncertain. Since only in a minority of cases are molecular techniques applicable, in this study we explored a multiparameter approach using phenotypic and functional characteristics to discriminate normal CD34 + cells from malignant ones.
Clinical Significance of CD200 and CD56 in Patients with Acute Lymphoblastic Leukemia
Scientific Journal for Damietta Faculty of Science, 2016
To analyze CD200 and CD56 expression by flow cytometry in acute lymphoblastic leukemia patients and whether their overexpression had prognostic impact individually and in combination with each other. Seventy newly diagnosed patients were assessed, of whom 27 were adult ALL and 43 pediatric ALL. Forty seven of 70 cases (67%) showed CD200 expression, and 7 cases (10%) showed CD56 expression but only 3 cases (4.3%) had expression for CD200+ and CD56+. Significance differences were found between CD200& CD56 expression in whole ALL patients group compared to control group (P<0.0001and P<0.001respectively). Splenomegaly and lower hemoglobin and platelet were more frequently observed in CD200+ patients whose also associated with significant increase of myeloid marker CD34 frequency. There were significant differences in overall survival (P= 0.042, P= 0.006) and disease-free survival (P= 0.005, P= 0.002) between the CD200+ and CD200-patients in total ALL patients and adult ALL. Whereas, in pediatric ALL OS of CD200+ patients 35.7% compared to 86.2% in CD200-, P= 0.032 but DFS showed nonsignificant difference (P=0.099). On the other hand, CD56+ patients had lower complete remission rate (14.3% vs. 63.5%, P= 0.018). CD56+ had significant influence on overall than those of CD56-(28.6% with mean 4.7 months vs. 41.1% with mean=11.8 months, P= 0.003) and disease free survival (40% with mean=6.26months vs. 54.9% with mean=14.16, P = 0.006). In respect to combination of two CDs positive had very inferior OS and DFS (mean =0.533 month and 0.150 month). In addition to, increased frequency of CD34 was associated with CD200+, CD56+ patients.
Study of Bone Marrow Lymphocyte Subset in Acute Myeloid Leukemia
Journal of Laboratory Physicians, 2021
Introduction Acute myeloid leukemia (AML) is a heterogenous disorder consisting of clonal expansion of myeloblasts. Tumor immunity plays an important part in the pathobiology of AML. Understanding the components of tumor immunity is important for understanding tumor pathogenesis and the principles of immunotherapy. Methods We studied 41 patients with AML, for total lymphocyte, CD4 positive helper T cells, CD8 positive cytotoxic T cells, and CD16/56 positive natural killer (NK) cells proportion. Quantification was done on bone marrow aspirate sample by flowcytometry. Whenever available, post induction bone marrow was also analyzed for the lymphocyte subset. Results No significant difference was noted in the percentage of blasts among the three risk categories: favorable, intermediate, and adverse. However, there was significant difference in the total lymphocyte among the risk stratification groups, being highest in the favorable group and lowest in the adverse group. CD8 positive cy...