Skip to main content

Detection of Terminal Transferase in Leukemia

  • Protocol
Flow Cytometry Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 91))

  • 1455 Accesses

Abstract

The mere presence of terminal deoxynucleotidyl transferase (TdT), a DNA polymerase, in leukemic cells provides no help in assigning these blast cells to a particular cell lineage (1). Differential levels of TdT gene transcription, however, result in diagnostically significant expression patterns of the enzyme with lower biochemical activity and weaker staining intensity by antibody recognition in myeloid as compared to lymphoid leukemia (24). One major advantage of measuring TdT by flow cytometry lies in its ability to objectively reflect staining intensities, a challenging task otherwise when one evaluates antibody staining under the microscope using the standard slide technique, thereby alleviating the need for cumbersome and expensive biochemical enzyme assays. The weak fluorescence staining of TdT-expressing myeloid leukemia cells, however, until recently has caused significant technical problems in the flow cytometric TdT detection, whereas several approaches have proven successful in the flow cytometric evaluation of TdT in the intensely staining lymphoid cells (3). Using optimal experimental conditions, the combined analysis of nuclear TdT and surface antigens in all types of leukemia now allows for the detection of minimal residual disease at levels as low as 0.02–0.5% of abnormal cells.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Paietta, E. (1995) Immunobiology of acute leukemia, in Neoplastic Diseases of the Blood, 3rd Ed (Wiernik, P. H, Canellos, G, Dutcher, J. P., and Kyle, R., eds.) Churchill Livingstone, New York, pp. 211–247

    Google Scholar 

  2. Paietta, E., Racevskis, J., Bennett, J. M., and Wiernik, P. H. (1993) Differential expression of terminal transferase (TdT) in acute lymphocytic leukemia expressing myeloid antigens and TdT positive acute myeloid leukemia as compared to myeloid antigen negative acute lymphocytic leukemia. Br. J Haematol. 84, 416–422.

    Article  CAS  PubMed  Google Scholar 

  3. Meenan, B., Heavey, C, Lichtenstein, A, Andersen, J., and Paietta, E (1996) Terminal transferase expression in the differential diagnosis of acute leukemias. Leukemia Lymphoma 22, 265–269.

    Article  CAS  PubMed  Google Scholar 

  4. Farahat, N., Lens, D., Morilla, R., Matutes, E., and Catovsky, D. (1995) Differential TdT expression in acute leukemia by flow cytometry: a quantitative study. Leukemia 9, 583–587.

    CAS  PubMed  Google Scholar 

  5. Bollum, F J. (1979) Terminal deoxynucleotidyl transferase as a hematopoietic cell marker. Blood 54, 1203–1215.

    CAS  PubMed  Google Scholar 

  6. Gore, S. D., Kastan, M. B., and Civic, C. I. (1991) Normal human bone marrow precursors that express terminal deoxynucleotidyl transferase include T-cell precursors and possible lymphoid stem cells. Blood 77, 1681–1690.

    CAS  PubMed  Google Scholar 

  7. Murray, M, Heavey, C, and Paietta, E. (1995) ORTHO Permeafix™ fixation is not suitable for the flow cytometric detection of nuclear terminal transferase in acute myeloid leukemia. Leukemia 9, 226, 227

    CAS  PubMed  Google Scholar 

  8. McCaffrey, R, Lillquist, A, Sallan, S, Cohen, E., and Osband, M. (1981) Clinical utility of leukemia cell terminal transferase measurements. Cancer Res. 41, 4814–4820.

    CAS  PubMed  Google Scholar 

  9. Hirata, M and Okamoto, Y (1987) Enumeration of terminal deoxynucleotidyl transferase positive cells in leukemia/lymphoma by flow cytometry. Leukemia Res 11, 509–518.

    Article  CAS  Google Scholar 

  10. Loftin, K. C, Reuben, J M, Dalton, W., Hersh, E M, and Sujansky, D (1986) Terminal transferase in leukemias by flow cytometry. Diag Immunol. 4, 165–169.

    CAS  Google Scholar 

  11. Slaper-Cortenbach, I C M., Admiraal, L G., Kerr, J. M., van Leeuwen, E. F., von dem Borne, A. E. G. Jr., and Tetteroo, P A. T. (1988) Flow-cytometric detection of terminal deoxynucleotidyl transferase and other intracellular antigens in combination with membrane antigens in acute lymphatic leukemias Blood 72, 1639–1644

    CAS  PubMed  Google Scholar 

  12. Bardales, R. H., Carrato, A., Fleischer, M., Schwartz, M K., and Koziner, B. (1989) Detection of terminal deoxynucleotidyl transferase (TdT) by flow cytometry in leukemic disorders J Histochem Cytochem 37, 509–513

    Article  CAS  PubMed  Google Scholar 

  13. Bettelheim, P., Paietta, E., Majdic, O, Gadner, H, Schwarzmeier, J., and Knapp, W (1982) Expression of a myeloid marker on TdT-positive acute lymphocytic leukemic cells evidence by double-fluorescence staining. Blood 60, 1392–1396

    CAS  PubMed  Google Scholar 

  14. Gore, S. D., Kastan, M B, Goodman, S. N, and Civin, C. I. (1990) Detection of minimal residual T-cell acute lymphoblastic leukemia by flow cytometry J Immunol Methods 132, 275–286.

    Article  CAS  PubMed  Google Scholar 

  15. Paietta, E., Meenan, B., Heavey, C, and Thomas, D. (1994) Detection of terminal transferase in acute myeloid leukemia by flow cytometry. Cytometry 16, 256–261

    Article  CAS  PubMed  Google Scholar 

  16. Drach, J., Gattringer, C, and Huber, H. (1991) Combined flow cytometric assessment of cell surface antigens and nuclear TdT for the detection of minimal residual disease in acute leukemia. Br. J. Haematol 77, 37–42.

    Article  CAS  PubMed  Google Scholar 

  17. Syrjala, M. T., Tiirikainen, M., Jansson, S.-E., and Krusius, T (1993) Flow cytometric analysis of terminal deoxynucleotidyl transferase. Hematopathology 99, 298–303.

    CAS  Google Scholar 

  18. Hallden, G., Andersson, U, Hed, J, and Johansson, S. G. O. (1989) A new membrane permeabilization method for the detection of intracellular antigens by flow cytometry. J Immunol Methods 124, 103–109.

    Article  CAS  PubMed  Google Scholar 

  19. Horvatinovich, J. M., Sparks, S. D, and Borowitz, M. J. (1994) Detection of terminal deoxynucleotidyl transferase by flow cytometry: a three color method. Cytometry 18, 228–230.

    Article  CAS  PubMed  Google Scholar 

  20. Pizzolo, G., Vincenzi, C, Nadali, G., Veneri, D., Vinante, F., Chilosi, M., Basso, G, Connelly, M. C, and Janossy, G (1994) Detection of membrane and intracellular antigens by flow cytometry following ORTHO Permeafix™ fixation. Leukemia 8, 672–676.

    CAS  PubMed  Google Scholar 

  21. Caldwell, C. W. (1994) Preservation of B-cell associated surface antigens by chemical fixation. Cytometry 16, 243–249.

    Article  CAS  PubMed  Google Scholar 

  22. Serke, S. (1995) Detection of terminal deoxynucleotidyl transferase by permeabilization of cells using a standard red blood cell lyse reagent. Cytometry 19, 189, 190.

    Article  CAS  PubMed  Google Scholar 

  23. Knapp, W., Strobl, H., and Majdic, O. (1994) Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis. Cytometry 18, 187–198.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Paietta, E. (1998). Detection of Terminal Transferase in Leukemia. In: Jaroszeski, M.J., Heller, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology™, vol 91. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-354-6:25

Download citation

  • DOI: https://doi.org/10.1385/0-89603-354-6:25

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-354-2

  • Online ISBN: 978-1-59259-214-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics