Identification of Individualized Minimal Residual Disease in Acute Myeloid Leukemia By Flow Cytometry and Its Application of Prognostic Significance
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Minimal Residual Disease
Clinical Significance
Immunophenotyping
The study of minimal residual disease (MRD) is an attempt to detect and define the significance of leukemia invisible to normal morphologic examination. In many circumstances the clinical significance of MRD detection is unclear, because the technical ability to detect and quantify it has outpaced studies demonstrating its clinical significance. The detection of minimal residual disease most consistently has been associated with relapse in acute lymphoblastic leukemia, t(15;17) acute myeloid leukemia, and chronic myeloid leukemia posttransplant, especially after T-cell depletion. But, in many types of leukemia, including acute myeloid leukemia and acute lymphoblastic leukemia, MRD can be detected in long-term remission patients without subsequent relapse. The study of MRD is evolving from detecting residual disease and predicting relapse to the study of the mechanisms that explain how minimum residual disease can coexist in a “cured” patient.
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This chapter discusses how minimal residual disease (MRD) is detected and managed in acute myeloid leukemia (AML) patients. The most commonly used techniques to detect residual leukemia in patients in complete remission (CR) are quantitative PCR (qPCR) and multicolor flow cytometry (MFC). While qPCR techniques have been extensively validated in multicenter efforts, and MRD detection using qPCR targets RNA or DNA sequences, which are derived from fusion genes, mutated genes, or genes overexpressed in AML cells, less formalized testing has been performed with MFC. For all major MRD markers, prognosis has been shown to be affected by the MRD level after the first course of chemotherapy. The applicability of MRD markers varies considerably. The majority of AML relapses occur within the first 2 years from diagnosis. That is not to say that relapse cannot occur later; but, on closer molecular characterization, late relapses will often turn out to be cases of secondary AML.
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Immunophenotyping
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Lymphoblast
Acute lymphocytic leukemia
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Multiple myeloma (MM) is a haematological malignancy characterized by the accumulation of monoclonal plasma cells in the bone marrow and remained incurable. Flow cytometry has been widely used in the detection of immunophenotype and minimal residual disease, diagnosis, monitoring and prognosis of MM. Normal plasma cells and malignant plasma cells can be distinguished according to different cell surface antigen expression. The clinical significane of many immune markes has been elucidated. However, the clinical significance of some phenotype remains controversial, the detection scheme and gating strategy are not unified. This review discusses the recent research progress on detection of MM immunophenotype and minimal residual disease by flow cytovetry.
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Minimal residual disease (MRD) following treatment is a robust prognostic marker in B lymphoblastic leukemia. However, the detection of MRD by flow cytometric immunophenotyping is technically challenging, and an automated method to detect MRD is therefore desirable. viSNE, a recently developed computational tool based on the t-Distributed Stochastic Neighbor Embedding (t-SNE) algorithm, has been shown to be capable of detecting synthetic "MRD-like" populations of leukemic cells created in vitro, but whether viSNE can facilitate the immunophenotypic detection of MRD in clinical samples has not been evaluated.
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The detection of minimal residual disease (MRD) is an important prognostic factor in childhood acute lymphoblastic leukemia (ALL) providing crucial information on the response to treatment and risk of relapse. However, the high cost of these techniques restricts their use in countries with limited resources. Thus, we prospectively studied the use of flow cytometry (FC) with a simplified 3-color assay and a limited antibody panel to detect MRD in the bone marrow (BM) and peripheral blood (PB) of children with ALL. BM and PB samples from 40 children with ALL were analyzed on days (d) 14 and 28 during induction and in weeks 24-30 of maintenance therapy. Detectable MRD was defined as > 0.01% cells expressing the aberrant immunophenotype as characterized at diagnosis among total events in the sample. A total of 87% of the patients had an aberrant immunophenotype at diagnosis. On d14, 56% of the BM and 43% of the PB samples had detectable MRD. On d28, this decreased to 45% and 31%, respectively. The percentage of cells with the aberrant phenotype was similar in both BM and PB in T-ALL but about 10 times higher in the BM of patients with B-cell-precursor ALL. Moreover, MRD was detected in the BM of patients in complete morphological remission (44% on d14 and 39% on d28). MRD was not significantly associated to gender, age, initial white blood cell count or cell lineage. This FC assay is feasible, affordable and readily applicable to detect MRD in centers with limited resources.
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Minimal residual disease (MRD) as a tool to monitor response to therapy is both a criterion for detailed risk stratification and an independent prognostic factor in childhood acute lymphoblastic leukemia (ALL). Immunological assays particularly flow cytometry (FC) are priority methods in MRD monitoring. Multicolor flow cytometry makes it possible to most fully characterize the immunophenotype of tumor B lymphoblasts and reveal leukemia-associated immunophenotypes not only according to the CD58 and CD38 antigens but also as an additional criterion of aberrancy. This allows you to identify and select individual criteria for further monitoring of minimal residual disease for each patient with ALL. The aim of this chapter is to compare immunophenotyping features of normal B-cell precursors and B-lymphoblasts in acute leukemia and to show possibilities of use of a leukemia-associated immunophenotype in monitoring of the MRD.
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Lymphoblast
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Objective:To investigate the expression of CD 56 on acute leukemia and its clinical manifestation.Methods:Immunophenotype of 92 cases with acute leukemia was measured by flow cytometry,the morphology,immunophenotype and clinic of CD 56 positive patients was analyzed.Results:15/92(16.3%)cases with acute leukemia expressed CD 56 ,one patient has been tesified to be myeloid/NK cell precursor acute leukemia,one patient has been testified to be blastic NK cell leukemia,one patient has been testified to be NK like T cell lymphoma/leukemia,12 patients are like to be acute myeloid leukemia with NK antigen expression or myeloid/NK cell acute leukemia.Conclusion:CD 56 positive acute leukemia is heterogeneous,its morphology,immunophenotype and clinical manifestation is different,they were associated with M 2?M 5?L 2;they had more infiltration out marrow and prognosis was poor.
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