Clinical Application Of Flow Cytometry Research Articles

Clinical Applications of Flow Cytometry in Cancer Immunotherapies: From Diagnosis to Treatments.

The scope of flow cytometry is rapidly expanding in the diagnosis of various cancers, and it is being used routinely as an aid in classifying leukemias and lymphomas. There are several applications of flow cytometry to enumerate tumorigenic anomalies in patients. The unusual distribution of cells in various locations, their DNA content, cell proliferation rate, dysregulated expression of several surface receptors, and expression of tumor antigens are some examples that can be characterized by using different flow cytometry-based techniques. For instance, the differential diagnosis between chronic lymphocytic leukemia (CLL) and various other mature B-cell neoplasms can be made by immunophenotyping in combination with absolute counting of numerous cellular subsets or by enumerating their percent distributions. Flow cytometry has several advantages over conventional techniques which include the ability to acquire a multiparametric data in a relatively shorter time and facilitate the comparative analysis of specific cellular subsets in an efficient manner.In addition to diagnosis, there are several other applications of flow cytometry in the management of various cancers which include treatment monitoring or even selecting a personalized precision-based immunotherapy in synch with advanced genetic tests to increase the chances of favorable prognosis and complete remission. The detection of chimeric antigen receptors (CARs) on various engineered effector cells can also be determined along with their specificity in engaging the targets. Furthermore, the assessment of numerous immunological parameters, their effector functions and potencies including the proliferation dynamics, cytokine secretion profiles, and activation efficiencies can also be measured before starting immunotherapies in patients.This chapter is a brief overview of flow cytometry applications in the diagnosis and treatment strategies of various cancers.

Immunophenotyping Investigation in Acute and Chronic Leukemias as a Gold Standard Test

Immunophenotyping of leukemias and other hematological neoplasms has become one of the most relevant clinical applications of flow cytometry which remains an indispensable tool for the diagnosis, classification, staging as well as hematologic malignancies monitoring. It can provide quantitative data on a number of cell parameters for a large number of cells in the region of cells thousands per second which making it a powerful diagnostic tool. If immunophenotyping is more sensitive significantly, specific and objective than morphological studying, the latter remains the gold standard criterion for creating and establishing the blast percentage in presentation and post treatment together. Moreover, it is important that interpreters of flow cytometric data have the phenotypes exact knowledge of diverse normal cell populations, which can be one of the best tool in recognize deviations from normal as well as are able to discuss the potential clinical significance and knowing look of the flow cytometric findings.

Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group.

This article discusses the rationale for inclusion of flow cytometry (FCM) in the diagnostic investigation and evaluation of cytopenias of uncertain origin and suspected myelodysplastic syndromes (MDS) by the European LeukemiaNet international MDS Flow Working Group (ELN iMDS Flow WG). The WHO 2016 classification recognizes that FCM contributes to the diagnosis of MDS and may be useful for prognostication, prediction, and evaluation of response to therapy and follow-up of MDS patients.

Diagnostic Utility of Immunophenotyping by Flow Cytometry for Diagnosis and Classification of Acute Leukemias in Tikur Anbessa Specialized Hospital, Addis Ababa, Ethiopia

Background: Immunophenotypic characterization of acute leukemia is an important clinical application of flow cytometry and has become a powerful tool contributing to proper diagnosis and classification. The aim of this study is to phenotype and classify acute leukemias by flow cytometry using commonly used markers for leukemia diagnosis.Methods: A total of 40 pediatric and adult patients diagnosed with acute leukemia were evaluated by flow cytometry with 17 surface and cytoplasmic markers known to be useful in discriminating different types of acute leukemia. These results were compared with classification results using standard morphological criteria.Results: 21 of 40 patients (52.5%) were classified as acute myeloblastic leukemia (AML) while 19 (47.5%) were identified as acute lymphoblastic leukemia (ALL). Of all the ALL cases, 10 of the 19 (52.6%) were B-ALL and 47.4% (9/19) were T-ALL based on flow cytometry. Markers of immaturity HLA-DR and CD34 antigens were co-expressed in 61% of AML cases and 33% of T-ALL cases, whereas CD34 was expressed in 50% of the B-ALL cases. cMPO and CD13 were the most expressed markers of AML, CD19 and cCD79a were present in all cases of B-ALL, and cytoplasmic CD3 and CD7 were positive on most all T-ALL cases. Discrimination of AML from ALL patients by flow cytometry was 80% concordant with traditional morphology. Notable discrepancies occurred in cases where leukemia cells expressed markers for more than one lineage.Conclusions: In the Ethiopia setting, flow cytometry represents a feasible and promising adjunctive diagnostic approach for acute leukemia.

Issue Highlights-July 2021.

Issue Highlights-July 2021.

Issue Highlights - May 2019.

Issue Highlights - May 2019.

The role of automated cytometry in the new era of cancer immunotherapy.

The introduction in the clinical practice of several new approaches to cancer immunotherapy has greatly increased the interest in analytical methodologies that can define the immunological profile of patients in the clinical setting. This requires huge effort to obtain reliable monitoring tools that could be used to improve the patient's clinical outcome. The clinical applications of flow cytometry (FCM) in oncology started with the measurement of DNA content for the evaluation of both ploidy and cell cycle profile as potential prognostic parameters in the majority of human solid cancer types. The availability of monoclonal antibodies widely broadened the spectrum of clinical applications of this technique, which rapidly became a fundamental tool for the diagnosis and prognosis of malignant hematological diseases. Among the emerging clinical applications of FCM, the study of minimal residual disease in hematological malignancies, the quantification of blood dendritic cells in various types of tumors, the study of metastatic spread in solid tumors throughout both the analysis of circulating endothelial progenitor cells and the identification and characterization of circulating tumor cells, all appear very promising. More recently, an advanced single cell analysis technique has been developed that combines the precision of mass spectrometry with the unique advantages of FCM. This approach, termed mass cytometry, utilizes antibodies conjugated to heavy metal ions for the analysis of cellular proteins by a mass spectrometer. It provides measurement of over 100 simultaneous cellular parameters in a single sample and has evolved from a promising technology to a high recognized platform for multi-dimensional single-cell analysis. Should a careful standardization of the analytical procedures be reached, both FCM and mass cytometry could effectively become ideal tools for the optimization of new immunotherapeutic approaches in cancer patients.

Fluorochromes for DNA Staining and Quantitation.

In these last few decades the great explosion of the molecular approaches has casted a little shadow on the DNA quantitative analysis. Nevertheless DNA cytochemistry represented a long piece of history in cell biology since the advent of the Feulgen reaction. This discovery was really the milestone of the emerging quantitative cytochemistry, and scientists from all over the world produced a very large literature on this subject. This first era of quantitation (histochemistry followed by cytochemistry) started by means of absorption measurements (histophotometry and cytophotometry). The successive introduction of fluorescence microscopy gave a great boost to quantitation, making easier and faster the determination of cell components by means of cytofluorometry. The development of flow cytometry further contributed to the importance of quantitative cytochemistry. At its beginning, the mission of flow cytometry was still DNA quantitation. For a decade the Feulgen reaction had been the reference methodology for both conventional and flow cytofluorometry; the advent of Shiff-type reagents contributed to expand the variety of possible fluorochromes excitable in the entire visible spectrum as well as in the ultraviolet region. The fluorescence scenario was progressively enriched by new probes among which are the intercalating dyes which made DNA quantitation simple and fast, thus spreading it worldwide. The final explosion of cytofluorometry was made possible by the availability of a large variety of probes directly binding DNA structure. In addition, immunofluorescence allowed to correlate the cell cycle-related DNA content to other cell markers. In the clinical application of flow cytometry, this promoted the introduction of multiparametric analyses aimed at describing the cytokinetic characteristics of a given cell subpopulation defined by a specific immunophenotype setting.

Recent advances on the prognostic value of immunophenotyping in multiple myeloma by flow cytometry

Clinical application of flow cytometry in multiple myeloma (MM) can be found in various dimensions, such as in differential diagnosis of malignant plasma cell disorder from reactive plasmacytosis, identification of the progression risk in MM, and in the detection of minimal residual disease. Flow cytometry-based clonality assessment with immuno-phenotyping encourages and enables the most stringent method of diagnosis and follow-up. The objective of this review is to summarize the recent information of the malignant plasma cell phenotypic profile of MM. The most comprehensive antigens, such as CD19, CD27, CD28, CD45, CD56 and CD117, play a significant role in the characterization of normal and malignant plasma cells. This review also focuses on the association of malignant phenotypic markers with chromosomal aberrations that identify the specific prognostic factors in MM.

Clinical Application of Flow Cytometry to Measure Anti-A/B IgG Antibody in ABO Incompatible Kidney Transplantation.

Clinical Application of Flow Cytometry to Measure Anti-A/B IgG Antibody in ABO Incompatible Kidney Transplantation.

Review of phenotypic markers used in flow cytometric analysis of MGUS and MM, and applicability of flow cytometry in other plasma cell disorders

Flow cytometric immunophenotyping is considered an indispensable tool for the diagnosis, classification and monitoring of disease in monoclonal gammopathies. The clinical sensitivity of flow cytometry is comparable with advanced molecular methods. Clinical application of flow cytometry in monoclonal gammopathies has various dimensions, such as differential diagnosis of malignant plasma cell disorder from reactive plasmacytosis, identifying the progression risk in monoclonal gammopathy of undetermined significance (MGUS) and asymptomatic multiple myeloma (MM), and minimal residual disease detection. Flow cytometry-based clonality assessment with immunophenotyping encourages and enables the most stringent method of diagnosis and follow-up. The objective of this review is to update the malignant plasma cells phenotypic profile of MGUS and MM. The most comprehensive antigens, such as CD19, CD27, CD28, CD45, CD56 and CD117, play a significant role in the characterization of normal and malignant plasma cells. Several research groups described the putative phenotype of myeloma cell progenitors, but no remarkable suggestion could be made because of disparity. This review also focuses on the association of malignant phenotypic markers and chromosomal aberrations that identify the specific prognostic features in monoclonal gammopathies.

Basic Theory and Clinical Applications of Flow Cytometry

Flow cytometry (FC) has become an important component in the diagnosis and monitoring of patients with a diverse array of diseases. While the basic principals underlying FC remain relatively unchanged, the technology and array of available reagents have continued to evolve, thus expanding the list of applications in laboratory medicine. This review provides an overview of the basic theory of FC including instrumentation, sample processing, and analysis. Additionally, current and future clinical applications of FC are discussed. Among these, applications of FC in solid organ and hematopoeitic stem-cell transplantation, transfusion medicine, hematopathology, and infectious diseases are presented. Finally, applications employed primarily in clinical research are presented in anticipation of their eventual use in the clinical laboratory setting.

Clinical Applications of Flow Cytometry for Nonneoplastic Disorders

Clinical Applications of Flow Cytometry for Nonneoplastic Disorders

Preface

Preface

Multidimensional flow cytometry immunophenotyping of hematologic malignancy.

Immunophenotyping of hematologic malignancies represents one of the most relevant clinical applications of flow cytometry. Classically, leukemic/lymphomatous cells have been considered to reflect the immunophenotypic characteristics of different precursors and mature healthy cells blocked at certain differentiation stages. Recently, accumulating evidence has shown that neoplastic cells display several aberrant phenotypic patterns. These aberrant phenotypes are believed to reflect genetic abnormalities present in pathologic cells, and recent data have shown that at least in acute leukemias, myelodysplastic syndromes, chronic lymphoproliferative disorders, and plasma cell dyscrasias, they may be present in almost all patients. The aim of this work is to review recent advances in flow cytometry and the role of gating strategies more useful in the identification and characterization of neoplastic cells of different hematologic malignancies.

Public Meeting and Workshop on ‘Safety issues pertaining to the clinical application of flow cytometry to human-derived cells’

Given the potential for a range of therapeutic procedures involving flow-sorted human cells, the US Center for Biologics Evaluation and Research (CBER) of the US Food and Drug Administration (FDA) realized that there were neither standards nor guidelines for the manipulation of human cells during cell sorting. Responding to a request from CBER, the International Society for Analytical Cytology (ISAC) agreed to co-sponsor a public workshop to solicit public comment regarding human-cell sorting. The purpose of the workshop was to provide a forum to obtain the current experience of laboratories and clinics involved in or planning such procedures, instrument manufacturers of cell sorters, as well as summaries of governmental regulatory issues that may affect future work in clinical cell-sorting. This public meeting and workshop was held April 20, 2001 on the NIH campus in Bethesda, MD, USA. It was convened to discuss the scientific and technological issues related to the development of voluntary safety protocols for clinical flow-cytometric sorting of human-derived cells. The workshop was designed to address the concerns of public health professionals, developing clinical protocols that use flow cytometry to sort cells for re-administration to humans, together with those of the manufacturers of these instruments. The two objectives of the workshop were to define the safety issues related to using flow cytometry to sort populations of human cells intended for clinical use, and to establish a working group to formulate voluntary protocols to be used by investigators intending to use flow cytometry in a clinical application. The following issues were specifically addressed: n The protection of the operator of the flow cytometer

Diagnostic advances in defining erythropoietic abnormalities and red blood cell diseases

The majority of clinical applications of flow cytometry begin with various approaches to remove red blood cells (RBCs) from the clinical sample. However, multiparameter cytometry has and will continue to contribute much to the understanding of the pathophysiology and diagnostic accuracy in the clinical evaluation of human diseases affecting erythroid cells. This review summarizes the diagnostic advances relating to erythroid cells in the areas of immunohematology, laboratory hematology, and infectious disease with particular emphasis on medical evaluation of the anemic patient and fetomaternal hemorrhage.

Clinical Applications of Flow Cytometry

Flow cytometry is an important analytical tool that has moved from research to the clinical laboratory in the past 20 years. Although still important in research, many applications have become routine clinical tests required for diagnosis, estimation of prognosis, and monitoring of treatment. In addition, new applications, such as minimal residual disease detection and multiple-drug resistance monitoring, are being evaluated for clinical testing.

Clinical applications of flow cytometry and cell immunophenotyping to companion animals (dog and cat).

Clinical applications of flow cytometry to certain diseases of the dog and cat are now possible. The utility of such applications for diagnosis, prognosis and follow-up are illustrated here by a number of examples: feline AIDS resulting from FIV infection, Leukocyte Adhesion Deficiency in Irish setters, deep pyoderma in German shepherds, Immune-mediated Thrombocytopenia, canine Systemic Lupus Erythematosus and Leishmaniasis, Leukemia and Lymphoma.

Clinically useful information provided by the flow cytometric immunophenotyping of hematological malignancies: current status and future directions.

At present, immunophenotyping of hematological malignancies represents one of the most relevant clinical applications of flow cytometry. In recent years, its use has extended from clinical research to diagnostic laboratories. The aim of this report is to critically review the type of information provided by the flow cytometric immunophenotyping of hematological malignancies and its clinical impact as well as to highlight its potential future applications. The currently available information, including that provided by different international consensus groups on the phenotypic characterization of hematologic malignancies, was reviewed. Additionally, recent reports on the immunophenotypic analysis of hematological malignancies published in hematology, oncology, pathology, immunology, and cell biology journals were also analyzed. A careful review of the literature showed that in spite of the well-established utility of immunophenotyping for the diagnosis, classification, prognostic stratification, and monitoring of hematological malignancies, only a small part of the information on the immunophenotypic characteristics of pathological hemopoietic cells has been used routinely. Specific and sensitive identification of neoplastic cells and their accurate enumeration and phenotypic characterization represent the major aims of these procedures. Similarities between leukemic and healthy cells allow the establishment of the lineage and maturation stage of the pathologic cells, this information being of great utility for the diagnosis, classification, and prognostic evaluation of different subtypes of hematological malignancies. On the other hand, the phenotypic aberrations displayed by leukemic cells could allow the selection of cases carrying specific genetic abnormalities in which further confirmatory molecular studies will be performed. The information provided by the flow cytometric immunophenotyping of hematological malignancies is of great clinical utility, with a major challenge for the near future being the standardization of technical procedures, data interpretation, and reporting.