Article

Craig FE, Foon KA. Flow cytometric immunophenotyping for hematologic neoplasms

Division of Hematopathology, Department of Pathology, University of Pittsburgh School of Medicine, PA, USA.
Blood (Impact Factor: 10.45). 05/2008; 111(8):3941-67. DOI: 10.1182/blood-2007-11-120535
Source: PubMed

ABSTRACT

Flow cytometric immunophenotyping remains an indispensable tool for the diagnosis, classification, staging, and monitoring of hematologic neoplasms. The last 10 years have seen advances in flow cytometry instrumentation and availability of an expanded range of antibodies and fluorochromes that have improved our ability to identify different normal cell populations and recognize phenotypic aberrancies, even when present in a small proportion of the cells analyzed. Phenotypically abnormal populations have been documented in many hematologic neoplasms, including lymphoma, chronic lymphoid leukemias, plasma cell neoplasms, acute leukemia, paroxysmal nocturnal hemoglobinuria, mast cell disease, myelodysplastic syndromes, and myeloproliferative disorders. The past decade has also seen refinement of the criteria used to identify distinct disease entities with widespread adoption of the 2001 World Health Organization (WHO) classification. This classification endorses a multiparametric approach to diagnosis and outlines the morphologic, immunophenotypic, and genotypic features characteristic of each disease entity. When should flow cytometric immunophenotyping be applied? The recent Bethesda International Consensus Conference on flow cytometric immunophenotypic analysis of hematolymphoid neoplasms made recommendations on the medical indications for flow cytometric testing. This review discusses how flow cytometric testing is currently applied in these clinical situations and how the information obtained can be used to direct other testing.

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    • "It has been widely used for many decades to monitor immune responses to vaccination and infection in bulk cell populations as well as to track the phenotypic and functional characteristics of antigen-specific cells, but it has also been largely applied in routine clinical settings for the diagnosis, prognosis and monitoring of disease. For example, it helps in the diagnosis and staging of patients with a hematological diseases [32]; for the detection of minimal residual disease (disease beyond the limit of morphological detection using conventional microscopy) [33]; for stem cell enumeration during immunosuppressive therapies [34]; in solid organ transplantation to evaluate T cell cross-match [35]; to monitor changes in cell populations after cardiopulmonary bypass surgery for the prediction of infections in risk patients [36]; in HIV for the determination of CD4 + T cell counts [37]; to predict hemolytic disease [38]; in primary [39] and secondary [40] immunodeficiencies; and largely used in blood transfusion [41]. The immunogenicity of vaccination and infection, and direct monitoring of the innate and adaptive immune responses can be measured by different methodologies. "

    Full-text · Article · Sep 2015 · Vaccine
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    • "Flow cytometry was performed using commercially available reagents (Becton- Dickinson Biosciences) following recommendations by Davis et al. [26] and Craig & Foon [27]. The panel of monoclonal antibodies used to determine acute leukemia lineage consisted of: CD45, CD34, HLA-DR, CD38, "
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    ABSTRACT: Introduction: Minimal residual disease is an important independent prognostic factor that can identify poor responders among patients with acute lymphoblastic leukemia. Objective: The aim of this study was to analyze minimal residual disease using immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements by conventional polymerase chain reaction followed by homo-heteroduplex analysis and to compare this with real-time polymerase chain reaction at the end of the induction period in children with acute lymphoblastic leukemia. Methods: Seventy-four patients diagnosed with acute lymphoblastic leukemia were enrolled. Minimal residual disease was evaluated by qualitative polymerase chain reaction in 57 and by both tests in 44. The Kaplan-Meier and multivariate Cox methods and the log-rank test were used for statistical analysis. Results: Nine patients (15.8%) were positive for minimal residual disease by qualitative polymerase chain reaction and 11 (25%) by real-time polymerase chain reaction considering a cut-off point of 1×10(-3) for precursor B-cell acute lymphoblastic leukemia and 1×10(-2) for T-cell acute lymphoblastic leukemia. Using the qualitative method, the 3.5-year leukemia-free survival was significantly higher in children negative for minimal residual disease compared to those with positive results (84.1%±5.6% versus 41.7%±17.3%, respectively; p-value=0.004). There was no significant association between leukemia-free survival and minimal residual disease by real-time polymerase chain reaction. Minimal residual disease by qualitative polymerase chain reaction was the only variable significantly correlated to leukemia-free survival. Conclusion: Given the difficulties in the implementation of minimal residual disease monitoring by real-time polymerase chain reaction in most treatment centers in Brazil, the qualitative polymerase chain reaction strategy may be a cost-effective alternative.
    Full-text · Article · Sep 2015 · Revista Brasileira de Hematologia e Hemoterapia
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    • "It has been widely used for many decades to monitor immune responses to vaccination and infection in bulk cell populations as well as to track the phenotypic and functional characteristics of antigen-specific cells, but it has also been largely applied in routine clinical settings for the diagnosis, prognosis and monitoring of disease. For example, it helps in the diagnosis and staging of patients with a hematological diseases [32]; for the detection of minimal residual disease (disease beyond the limit of morphological detection using conventional microscopy) [33]; for stem cell enumeration during immunosuppressive therapies [34]; in solid organ transplantation to evaluate T cell cross-match [35]; to monitor changes in cell populations after cardiopulmonary bypass surgery for the prediction of infections in risk patients [36]; in HIV for the determination of CD4 + T cell counts [37]; to predict hemolytic disease [38]; in primary [39] and secondary [40] immunodeficiencies; and largely used in blood transfusion [41]. The immunogenicity of vaccination and infection, and direct monitoring of the innate and adaptive immune responses can be measured by different methodologies. "
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    ABSTRACT: The immune system is a network of specialized cell types and tissues that communicates via cytokines and direct contact, to orchestrate specific types of defensive responses. Until recently, we could only study immune responses in a piecemeal, highly focused fashion, on major components like antibodies to the pathogen. But recent advances in technology and in our understanding of the many components of the system, innate and adaptive, have made possible a broader approach, where both the multiple responding cells and cytokines in the blood are measured. This systems immunology approach to a vaccine response or an infection gives us a more holistic picture of the different parts of the immune system that are mobilized and in turn, may allow us to better understand the mechanisms of such responses, as well as to predict vaccine efficacy in different populations well in advance of efficacy studies. Here we summarize the different technologies and methods and discuss how they can inform us about the differences between diseases and vaccines, and how they can greatly accelerate vaccine development. Copyright © 2015. Published by Elsevier Ltd.
    Full-text · Article · Jul 2015 · Vaccine
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