Advances in Cell biology, vol. 32, 2005, issue 1

Summary: Antibodies are protein particles capable of specific antigen binding. Policlonal antibodies comprise of a group of antibodies that recognize different epitopes of the same antigen. Monoclonal antibodies recognize and bind to only one specific epitope. Both types of antibodies have their applications in diagnostic techniques, however only monoclonal antibodies are used in flow cytometry. Conjugation of monoclonal antibodies with fluorescent dyes enables the detection of antigens of different cell types. Contemporary flow cytometers can simultaneously detect more than ten different fluorescences, which gives the possibility of simultaneous assessment of many different antigens of the same cell.

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Summary: Immunophenotyping is a significant method for hematooncology diseases diagnosis and treatment. The knowledge about normal hematopoiesis enables the estimation of aberrant antigens patterns in pathological specimen. In this paper the authors are presenting immunophenotypic differentiation patterns in normal hematopoiesis. A three-, four-, and six color flow cytometry analysis scheme of antigens expression have also been proposed.

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Immunophenotype of Mature Hematopoietic Cells – Primary and Secondary Immunodeficiences

Summary: The leukocytes characteristics from peripheral blood and lymph nodes including age-related changes were shown. Primary immunodeficiencies were described based on quantitative changes within lymphocytes T and B populations assayed with flow cytometry. Immunodeficiences with diagnosis based on analysis of peripheral blood and lymph node cells' immunophenotypes were discussed.

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Summary: Modern diagnostics of hematological diseases is a combination of different laboratory techniques. Flow cytometry is an indispensable tool for the diagnosis of leukemia and lymphoma used in addition to classical cytology, cytochemistry, cytogenetics and molecular biology. The advantage of flow cytometry for diagnosis in hematology is a multiparameter assessment of antigens on a single cell level. Specifically in cases of leukemia flow cytomerty allows the determination lineage and maturation status of the malignant cells, and so the classification of the subtype of leukemia, identification of aberrant antigen expression and makes possible further detection of minimal residual disease. This article includes the immunological classification of acute leukemias, shows the examples of different monoclonal antibody panels that can be used for leukemia diagnostics and presents a selection of case reports taken from the routine work in our Department of Hematology. This article could be of practical help for people commencing immunophenotyping of acute leukemias.

Summary: Lymphomas are tumors of the lymphatic system, but its cells, mainly lymphocytes constitute morphological substrate of the immune system. Lymphocytes are characterized by the expression of several antigens manifested in subsequent stages of development. These, so called differentiation antigens grouped in CD classification are also expressed in lymphomas. They may be traced by means of various methods, and in particular, by flow cytometry. The latter has not only diagnostic function, but also permits to detect residual disease and allows for monitoring of neoplastic process in the course of anti-tumor therapy. Diagnostics of lymphomas is still based on histopathological assessment of tissue sections. Flow cytometry constitutes however valuable complement, providing quantitative data of positive cells, percent of proliferating ones, DNA ploidy, and above all, fast response. It does not however, provide information about topography of lesion. Cells for flow cytometry may originate not only from blood or bone marrow, but also from oligobiopsy of tumor or lymph node. Marked and steadily growing number of known differentiation antigens creates vast possibilities for the evaluation of not only B-cell but also T-cell and NK-cell tumors. Technology of flow cytometry, based on the assessment of several thousands of living cells in the laser beam, subjected previously to the reaction with fluorochrome labelled monoclonal antibodies – is relatively simple and permits to obtain comprehensive results within couple of hours. In summary, the data presented, with the examples of cytometric diagrams, show unequivocally, that parallel use of flow cytometry with morphological assessment of tissue section may constitute „gold standard” in diagnostics and monitoring of this group of malignancies.

Detection of Minimal Residual Disease in Acute Lymphoblastic Leukemia in Children

Summary: Detection of minimal residual disease (MRD) in acute lymphoblastic leukemia with flow cytometry is based on precise description of leukemia cell at diagnosis including aberrant expression of determinants. The important clinical significance of MRD is in time of obtaining the first remission of acute leukemia. The problems with MRD assay and interpretation of results in acute lymphoblastic B and T leukemia are discussed.

Summary: Flow cytometry is applied to plant research for the estimation of ploidy, cell cycle activity, endoreplication level and genome size. Almost all tissues and organs can be analyzed, including those originated from in vitro culture. Protocols for sample preparation and flow cytometric analyses of different plant tissues are described, and recipes are given for isolation/staining buffers.