Flow-Cytometric Detection of Terminal Deoxynucleotidyl Transferase and Other Intracellular Antigens in Combination With Membrane Antigens in Acute Lymphatic Leukemias

• Published in: Blood Vol. 72; no. 5; pp. 1639 - 1644
• Main Authors: Slaper-Cortenbach, Ineke CM, Admiraal, Lucia G., Kerr, Jane M., Leeuwen, Eleonore F. van, Borne, Albert E.G. Kr. von dem, Tetteroo, Pedro A.T.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 01-11-1988; The Americain Society of Hematology
• Subjects: Antigens - analysis; Antigens, Differentiation - analysis; Antigens, Neoplasm - analysis; Antigens, Surface - analysis; Biological and medical sciences; Biomarkers, Tumor - analysis; Cell Membrane Permeability; DNA Nucleotidylexotransferase - blood; Fixatives; Flow Cytometry; Fluorescent Antibody Technique; Hematologic and hematopoietic diseases; Humans; Leukemia, Lymphocytic, Chronic, B-Cell - enzymology; Leukemia, Lymphocytic, Chronic, B-Cell - immunology; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Medical sciences; Precursor Cell Lymphoblastic Leukemia-Lymphoma - enzymology; Precursor Cell Lymphoblastic Leukemia-Lymphoma - immunology; Human; Flow cytometry; Residual tumor; Histological preparation; Hemopathy; Monoclonal antibody; Antigen; DNA nucleotidylexotransferase; Screening; Plasma membrane; Acute lymphocytic leukemia; Immunofluorescence; Cytoplasm
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V72.5.1639.1639

Development of a new fixation procedure allowed flow-cytometric analysis of nuclear and other intracellular antigens in acute lymphatic leukemia (ALL). A short fixation of the cells with buffered formaldehyde acetone (BFA) rendered the cell membrane permeable, allowing the monoclonal antibodies (MoAbs) to penetrate the cell. Through this method, a rapid analysis of intracellular antigens, specific for acute lymphatic leukemia [such as terminal deoxynucleotidyl transferase (TdT), immunoglobulin M (IgM) heavy chain, and antigens recognized by the CD22 or CD3 MoAbs) was performed by flow cytometry. The surface antigens remained intact after this fixation procedure, enabling simultaneous detection of membrane and intracellular antigens. The binding of biotinylated antibodies against several B- and T-lymphoid membrane antigens was detected with streptavidin-phycoerythrin (red fluorescence), whereas the intracellular antigens were stained with FITC-labeled polyclonal antibodies, or indirectly with FITC-labeled goat anti-mouse IgG (green fluorescence). Through this combination of markers, minor cell populations can be detected and a rapid and quantitative immunodiagnosis can be performed.