The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Alberto Orfao - One of the best experts on this subject based on the ideXlab platform.
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euroflow resetting leukemia and lymphoma Immunophenotyping basis for companion diagnostics and personalized medicine
Leukemia, 2012Co-Authors: J J M Van Dongen, Alberto OrfaoAbstract:Laboratory diagnostics in patients with a hematological malignancy has three major applications: establishing the diagnosis, prognostic classification and evaluation of treatment effectiveness.1, 2 Immunophenotyping is currently recognized to provide essential information for all three applications.2, 3, 4, 5, 6, 7, 8, 9 Expression of individual immunophenotypic markers was initially assessed by microscopic techniques, but since the 90's multiparameter flow cytometric Immunophenotyping has become the technique of choice, as it is the sole technique that fulfills the requirements for high speed, broad applicability at diagnosis and during follow-up, and accurate focusing on the malignant cell population using membrane-bound and intracellular proteins as targets.9, 10, 11, 12, 13, 14 Despite the objectivity of flow cytometric measurements, flow cytometry is perceived as a technique that is highly dependent on expertise and is regarded to have limited reproducibility in multicenter studies.15, 16, 17, 18 This probably relates to the increasing number of antibodies and fluorochromes that are used and the corresponding progressively larger complexity of the multivariate data analyses of both major and minor cell populations, together with limited standardization of the laboratory procedures and instrument settings. In this regard, the weakest points of multiparameter flow cytometry relate to: (i) the design of the panels of markers to be applied, (ii) the evaluation of new versus ‘classical' markers, (iii) the analysis of the data obtained from the flow cytometric measurements and (iv) interpretation of the results. In addition, it is a technological field that is continuously evolving, but where many traditional procedures are still in use, for example, for data analysis. Whereas industry invested significantly in developing and implementing further innovation of the flow cytometry instruments, the innovation in Immunophenotyping reagents and in software for analysis of the progressively larger and complex data sets was much more limited or virtually absent, particularly in the area of leukemia and lymphoma typing. Consequently, we concluded in 2005 that major innovations are required to adequately advance the field of flow cytometric Immunophenotyping. Therefore, we initiated the European Union (EU)-supported EuroFlow Consortium. The original objectives of the EuroFlow Consortium were: the development and evaluation of novel antibodies, the introduction of novel immunobead technology, the development of novel flow cytometry software tools and data analysis approaches for recognition of complex immunophenotypic patterns, and the design of novel multicolor immunostaining protocols and carefully balanced antibody panels. In this editorial, we critically comment on the most relevant aspects of the EuroFlow activities through a series of frequently asked questions from the field.
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multiparameter flow cytometry quantification of bone marrow plasma cells at diagnosis provides more prognostic information than morphological assessment in myeloma patients
Haematologica, 2009Co-Authors: Bruno Paiva, Mariabelen Vidriales, Alberto Orfao, Jose Isabel Juan Perez, Gema Mateo, Maria Angeles Montalban, Mariavictoria Mateos, Joan Blade, Juan Jose Lahuerta, Jesus San F MiguelAbstract:Quantification of bone marrow plasma cells in multiple myeloma patients using conventional morphology is of limited prognostic value, while the merit of multiparameter flow cytometry Immunophenotyping is still considered unproven. Here we compare the bone marrow plasma cell counts obtained by morphology and multiparameter flow cytometry and explore the potential prognostic impact of both techniques in 765 newly diagnosed, uniformly treated multiple myeloma patients. Although multiparameter flow cytometry generally yields lower plasma cell counts (median percentage of 11% vs. 40%, respectively; p<0.001), there is a significant positive correlation between the two techniques (R =0.46, p<0.001). Regarding prognosis, multivariate analysis selected the bone marrow plasma cell counts obtained by multiparameter flow cytometry as an independent prognostic factor for overall survival (p=0.007), supporting the incorporation of multiparameter flow cytometry Immunophenotyping into the routine diagnostic evaluation of multiple myeloma patients and validating the clinical utility of bone marrow plasma cell counting by multiparameter flow cytometry approaches. (clinicaltrials.gov identifier: NCT00560053).
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flow cytometric Immunophenotyping of cerebrospinal fluid
Current protocols in immunology, 2008Co-Authors: Jaco Kraan, Anna Porwit, J W Gratama, Corinne Haioun, Alberto Orfao, Anne Plonquet, Sandra Quijano, Maryalice Stetlerstevenson, Dolores Subira, W E WilsonAbstract:Leptomeningeal disease is an important adverse complication occurring in patients with B and T cell lymphomas and acute leukemias of lymphoid and myeloid origin. Recent reports suggest that multiparameter flow cytometry immunophenotypic assessment of spinal fluid samples could improve the efficiency of detection of CNS involvement, due to its high specificity and greater sensitivity. However, spinal fluid samples are frequently paucicellular with a rapidly decreasing cell viability. Staining of spinal fluid therefore requires dedicated sample storage/transport, staining, and preparation protocols. The Basic Protocol in this unit outlines a consensus multiparameter (3- to 8-color) flow cytometry immunophenotypic protocol for the evaluation of CNS involvement of cerebrospinal fluid (CSF) samples by neoplastic cells. A Support Protocol describing the simultaneous assessment of surface and cytoplasmic antigens is also provided. Finally, in the Alternate Protocol, we describe a method to calculate absolute numbers of both normal and pathological cell subpopulations by adding counting beads to the assay.
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Immunophenotyping of acute leukemias and myelodysplastic syndromes
Cytometry Part A, 2004Co-Authors: Alberto Orfao, Francisco Jose Ortuno, Maria De Santiago, Antonio Lopez, Jesus San F MiguelAbstract:Alberto Orfao,* Francisco Ortuno, Maria de Santiago, Antonio Lopez, and Jesus San Miguel Servicio General de Citometria, Universidad de Salamanca, Salamanca, Spain Centro de Investigacion del Cancer y Departamento de Medicina, Universidad de Salamanca, Salamanca, Spain Servicio de Hematologia y Oncologia Medica, Hospital General Universitario J.M. Morales Meseguer, Murcia, Spain Servicio de Hematologia, Hospital Universitario de Salamanca, Salamanca, Spain
Luigi Del Vecchio - One of the best experts on this subject based on the ideXlab platform.
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flow cytometry Immunophenotyping for the evaluation of bone marrow dysplasia
Cytometry Part B-clinical Cytometry, 2011Co-Authors: Matteo G Della Porta, Francesco Lanza, Luigi Del VecchioAbstract:The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This classification provides clinicians with a useful tool for defining the different subtypes of MDS and determining individual prognosis. The WHO proposal has raised some concern regarding minimal diagnostic criteria particularly in patients with normal karyotype without robust morphological markers of dysplasia (such as ring sideroblasts or excess of blasts). Therefore, there is clearly a need to refine the accuracy to detect marrow dysplasia. Flow cytometry (FCM) Immunophenotyping has been proposed as a tool to improve the evaluation of marrow dysplasia. Rationale for the application of FCM in the diagnostic work up of MDS is that Immunophenotyping is an accurate method for quantitative and qualitative evaluation of hematopoietic cells and that MDS have been found to have abnormal expression of several cellular antigens. To become clinically applicable, FCM analysis should be based on parameters with sufficient specificity and sensitivity, data should be reproducible between different operators and the results should be easily understood by clinicians. In this report, we reviewed the most relevant progresses in detection of marrow dysplasia by FCM in MDS as defined by WHO criteria.
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flow cytometry Immunophenotyping for the evaluation of bone marrow dysplasia
Cytometry Part B-clinical Cytometry, 2011Co-Authors: Matteo G Della Porta, Francesco Lanza, Luigi Del VecchioAbstract:The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This classification provides clinicians with a useful tool for defining the different subtypes of MDS and determining individual prognosis. The WHO proposal has raised some concern regarding minimal diagnostic criteria particularly in patients with normal karyotype without robust morphological markers of dysplasia (such as ring sideroblasts or excess of blasts). Therefore, there is clearly a need to refine the accuracy to detect marrow dysplasia. Flow cytometry (FCM) Immunophenotyping has been proposed as a tool to improve the evaluation of marrow dysplasia. Rationale for the application of FCM in the diagnostic work up of MDS is that Immunophenotyping is an accurate method for quantitative and qualitative evaluation of hematopoietic cells and that MDS have been found to have abnormal expression of several cellular antigens. To become clinically applicable, FCM analysis should be based on parameters with sufficient specificity and sensitivity, data should be reproducible between different operators and the results should be easily understood by clinicians. In this report, we reviewed the most relevant progresses in detection of marrow dysplasia by FCM in MDS as defined by WHO criteria. © 2011 International Clinical Cytometry Society
Matteo G Della Porta - One of the best experts on this subject based on the ideXlab platform.
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harmonemia a universal strategy for flow cytometry Immunophenotyping a european leukemianet wp10 study
Leukemia, 2016Co-Authors: Francis Lacombe, E Bernal, David Bloxham, S Couzens, Matteo G Della Porta, Ulf Johansson, Wolfgang Kern, M G Macey, Thomas Matthes, R MorillaAbstract:Harmonemia: a universal strategy for flow cytometry Immunophenotyping—A European LeukemiaNet WP10 study
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flow cytometry Immunophenotyping for the evaluation of bone marrow dysplasia
Cytometry Part B-clinical Cytometry, 2011Co-Authors: Matteo G Della Porta, Francesco Lanza, Luigi Del VecchioAbstract:The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This classification provides clinicians with a useful tool for defining the different subtypes of MDS and determining individual prognosis. The WHO proposal has raised some concern regarding minimal diagnostic criteria particularly in patients with normal karyotype without robust morphological markers of dysplasia (such as ring sideroblasts or excess of blasts). Therefore, there is clearly a need to refine the accuracy to detect marrow dysplasia. Flow cytometry (FCM) Immunophenotyping has been proposed as a tool to improve the evaluation of marrow dysplasia. Rationale for the application of FCM in the diagnostic work up of MDS is that Immunophenotyping is an accurate method for quantitative and qualitative evaluation of hematopoietic cells and that MDS have been found to have abnormal expression of several cellular antigens. To become clinically applicable, FCM analysis should be based on parameters with sufficient specificity and sensitivity, data should be reproducible between different operators and the results should be easily understood by clinicians. In this report, we reviewed the most relevant progresses in detection of marrow dysplasia by FCM in MDS as defined by WHO criteria.
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flow cytometry Immunophenotyping for the evaluation of bone marrow dysplasia
Cytometry Part B-clinical Cytometry, 2011Co-Authors: Matteo G Della Porta, Francesco Lanza, Luigi Del VecchioAbstract:The pathological hallmark of myelodysplastic syndromes (MDS) is marrow dysplasia, which represents the basis of the WHO classification of these disorders. This classification provides clinicians with a useful tool for defining the different subtypes of MDS and determining individual prognosis. The WHO proposal has raised some concern regarding minimal diagnostic criteria particularly in patients with normal karyotype without robust morphological markers of dysplasia (such as ring sideroblasts or excess of blasts). Therefore, there is clearly a need to refine the accuracy to detect marrow dysplasia. Flow cytometry (FCM) Immunophenotyping has been proposed as a tool to improve the evaluation of marrow dysplasia. Rationale for the application of FCM in the diagnostic work up of MDS is that Immunophenotyping is an accurate method for quantitative and qualitative evaluation of hematopoietic cells and that MDS have been found to have abnormal expression of several cellular antigens. To become clinically applicable, FCM analysis should be based on parameters with sufficient specificity and sensitivity, data should be reproducible between different operators and the results should be easily understood by clinicians. In this report, we reviewed the most relevant progresses in detection of marrow dysplasia by FCM in MDS as defined by WHO criteria. © 2011 International Clinical Cytometry Society
Jesus San F Miguel - One of the best experts on this subject based on the ideXlab platform.
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multiparameter flow cytometry quantification of bone marrow plasma cells at diagnosis provides more prognostic information than morphological assessment in myeloma patients
Haematologica, 2009Co-Authors: Bruno Paiva, Mariabelen Vidriales, Alberto Orfao, Jose Isabel Juan Perez, Gema Mateo, Maria Angeles Montalban, Mariavictoria Mateos, Joan Blade, Juan Jose Lahuerta, Jesus San F MiguelAbstract:Quantification of bone marrow plasma cells in multiple myeloma patients using conventional morphology is of limited prognostic value, while the merit of multiparameter flow cytometry Immunophenotyping is still considered unproven. Here we compare the bone marrow plasma cell counts obtained by morphology and multiparameter flow cytometry and explore the potential prognostic impact of both techniques in 765 newly diagnosed, uniformly treated multiple myeloma patients. Although multiparameter flow cytometry generally yields lower plasma cell counts (median percentage of 11% vs. 40%, respectively; p<0.001), there is a significant positive correlation between the two techniques (R =0.46, p<0.001). Regarding prognosis, multivariate analysis selected the bone marrow plasma cell counts obtained by multiparameter flow cytometry as an independent prognostic factor for overall survival (p=0.007), supporting the incorporation of multiparameter flow cytometry Immunophenotyping into the routine diagnostic evaluation of multiple myeloma patients and validating the clinical utility of bone marrow plasma cell counting by multiparameter flow cytometry approaches. (clinicaltrials.gov identifier: NCT00560053).
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Immunophenotyping of acute leukemias and myelodysplastic syndromes
Cytometry Part A, 2004Co-Authors: Alberto Orfao, Francisco Jose Ortuno, Maria De Santiago, Antonio Lopez, Jesus San F MiguelAbstract:Alberto Orfao,* Francisco Ortuno, Maria de Santiago, Antonio Lopez, and Jesus San Miguel Servicio General de Citometria, Universidad de Salamanca, Salamanca, Spain Centro de Investigacion del Cancer y Departamento de Medicina, Universidad de Salamanca, Salamanca, Spain Servicio de Hematologia y Oncologia Medica, Hospital General Universitario J.M. Morales Meseguer, Murcia, Spain Servicio de Hematologia, Hospital Universitario de Salamanca, Salamanca, Spain
Cherie H. Dunphy - One of the best experts on this subject based on the ideXlab platform.
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applications of flow cytometry and immunohistochemistry to diagnostic hematopathology
Archives of Pathology & Laboratory Medicine, 2004Co-Authors: Cherie H. DunphyAbstract:c Objective.—Diagnostic hematopathology depends on the applications of flow cytometric Immunophenotyping and immunohistochemical Immunophenotyping combined with the cytomorphology and histologic features of each case. Select cases may require additional ancillary cytogenetic and molecular studies for diagnosis. The purpose of this review is to focus on the applications of flow cytometric and immunohistochemical Immunophenotyping of paraffin-embedded tissue to diagnostic hematopathology. Advantages and disadvantages of these techniques are examined. Data Sources.—The literature is extensively reviewed (PubMed 1985‐2003) with an emphasis on the most recent applications and those that are most useful clinically, both diagnostically and prognostically. Study Selection.—Studies were selected based on statistically significant results in large studies with reported adequate clinical follow-up. Data Extraction.—The methodology was reviewed in the selected studies to ensure reliable comparison of reported data. Data Synthesis.—Flow cytometric Immunophenotyping offers the sensitive detection of antigens for which antibodies may not be available for paraffin immunohistochemical Immunophenotyping. However, paraffin immunohistochemical Immunophenotyping offers preservation of architecture and evaluation of expression of some proteins, which may not be available by flow cytometric Immunophenotyping. These techniques should be used as complimentary tools in diagnostic hematopathology. Conclusions.—There are extensive applications of flow cytometric and immunohistochemical Immunophenotyping to diagnostic hematopathology. As cytogenetic and molecular findings evolve in diagnostic hematopathology, there may be additional applications of flow cytometric and immunohistochemical Immunophenotyping to this field of pathology. (Arch Pathol Lab Med. 2004;128:1004‐1022)
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applications of flow cytometry and immunohistochemistry to diagnostic hematopathology
Archives of Pathology & Laboratory Medicine, 2004Co-Authors: Cherie H. DunphyAbstract:c Objective.—Diagnostic hematopathology depends on the applications of flow cytometric Immunophenotyping and immunohistochemical Immunophenotyping combined with the cytomorphology and histologic features of each case. Select cases may require additional ancillary cytogenetic and molecular studies for diagnosis. The purpose of this review is to focus on the applications of flow cytometric and immunohistochemical Immunophenotyping of paraffin-embedded tissue to diagnostic hematopathology. Advantages and disadvantages of these techniques are examined. Data Sources.—The literature is extensively reviewed (PubMed 1985‐2003) with an emphasis on the most recent applications and those that are most useful clinically, both diagnostically and prognostically. Study Selection.—Studies were selected based on statistically significant results in large studies with reported adequate clinical follow-up. Data Extraction.—The methodology was reviewed in the selected studies to ensure reliable comparison of reported data. Data Synthesis.—Flow cytometric Immunophenotyping offers the sensitive detection of antigens for which antibodies may not be available for paraffin immunohistochemical Immunophenotyping. However, paraffin immunohistochemical Immunophenotyping offers preservation of architecture and evaluation of expression of some proteins, which may not be available by flow cytometric Immunophenotyping. These techniques should be used as complimentary tools in diagnostic hematopathology. Conclusions.—There are extensive applications of flow cytometric and immunohistochemical Immunophenotyping to diagnostic hematopathology. As cytogenetic and molecular findings evolve in diagnostic hematopathology, there may be additional applications of flow cytometric and immunohistochemical Immunophenotyping to this field of pathology. (Arch Pathol Lab Med. 2004;128:1004‐1022)
