Detection of terminal transferase in acute myeloid leukemia by flow cytometry

Elisabeth M. Paietta, B. Meenan, C. Heavey, D. Thomas

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The purpose of this study was twofold: (1) to develop an optimized, reliable method for the flow cytometric analysis of the intranuclear DNA polymerase, terminal deoxynucleotidyl transferase (TdT) in acute myeloid leukemia, and (2) to establish the usefulness of a novel, fluorescein-isothiocyanate conjugated monoclonal anti-TdT antibody (HT-6) in double-fluorescence staining for surface antigens in the characterization of leukemic cells. Inclusion of an aldehyde blocking buffer in the staining protocol reduced background fluorescence sufficiently to allow for the detection of the low-level fluorescent TdT+ myeloblasts. When admixed to normal peripheral blood mononuclear cells, 0.4-0.5% of HLA-DR+ or myeloid surface antigen+, TdT+ double-stained myeloblasts could be reliably detected above background levels. Flow cytometric TdT measurements using the HT-6 antibody in 55 patients with TdT+ acute lymphocytic or myelocytic leukemia or blast crisis of chronic myelogenous leukemia were equal or superior to the results obtained with a mixture of monoclonal anti-TdT antibodies (anti-HTDT-Mix) and comparable to those obtained by the conventional slide method employing polyclonal rabbit anti-human TdT antiserum. This flow cytometric TdT determination in combination with surface antigen staining using a novel anti-TdT monoclonal antibody( HT-6) allows for the recognition of minimal leukemic blast cells during clinical remission in acute myeloid leukemia.

Original languageEnglish (US)
Pages (from-to)256-261
Number of pages6
JournalCytometry
Volume16
Issue number3
DOIs
StatePublished - 1994

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DNA Nucleotidylexotransferase
Transferases
Acute Myeloid Leukemia
Flow Cytometry
Surface Antigens
Granulocyte Precursor Cells
Staining and Labeling
Fluorescence
Blast Crisis
Antibodies
HLA-DR Antigens
DNA-Directed DNA Polymerase
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Fluorescein
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Aldehydes
Immune Sera
Anti-Idiotypic Antibodies
Blood Cells
Buffers

Keywords

  • AML
  • double staining
  • flow cytometry
  • monoclonal anti-TdT antibody

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Detection of terminal transferase in acute myeloid leukemia by flow cytometry. / Paietta, Elisabeth M.; Meenan, B.; Heavey, C.; Thomas, D.

In: Cytometry, Vol. 16, No. 3, 1994, p. 256-261.

Research output: Contribution to journalArticle

Paietta, Elisabeth M. ; Meenan, B. ; Heavey, C. ; Thomas, D. / Detection of terminal transferase in acute myeloid leukemia by flow cytometry. In: Cytometry. 1994 ; Vol. 16, No. 3. pp. 256-261.
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