Experience in theuse of polymerase chain reaction for determining T-cellclonality


Aim. To distinguish T-cell lymphomas and reactive T-cell proliferation it is important to confirm the
ability of T-cells to be cloned. Conventional histological and immunophenotypic methods fail to determine
the ability of T-cells to be cloned. An experience in the use of detection of T-cell receptor gene
gamma-chain (TCRy) rearrangement for determining T-cellular clonality is described.
Material and methods. Polymerase chain reaction (PCR) and single strand conformational polymorphism
(SSCP) were used to determine T-cell clonality. Twenty healthy donors, 28 patients with T-lymphomas,
and 26patients with various non-T-cell lymphoproliferative disorders or reactive processes
were studied.
Results. T-cell monoclonality was detected in 23/28 (82%) T-cell lymphoma cases, whereas in all the
samples from normal subjects a polyclonal pattern of rearrangements TCRy was found. The sensitivity
of the method was estimated as 2,5%, 7%, and 10% was demonstrated for bone marrow, spleen, and
peripheral blood, respectively.
Conclusion. PCR-SSCP for TCRy was found to be a useful supplement to routine histological and immunophenotypic
methods in the diagnosis of T-cell lymphomas.


  1. Воробьев А. И., Кременецкая А. М., Харазшивили Д. Опухоли лимфатической системы. Гематол. и трансфузиол 2000; 3: 1-14.
  2. WHO. Classification of neoplasms of the hematopoietic amlymphoid tissues / Harris N. L., Jaffe E. S., Diebold J. et al Airlie House, Virginia 1997.
  3. Никитин E. А., Левина E. А., Судариков А. Б. Роль молекулярных методов в диагностике и мониторинге лимфопролиферативных заболеваний. Гематол. и трансфузиол 2001; 3: 19-26.
  4. Signoretti S., Murphy M., Cangi M. G. et al. Detection of clonal T-cell receptor у gene rearrangements in paraffm-embeddectissue by polymerase chain reaction and nonradioactive singlestrand conformational polymorphism analysis. Am. J. Pathol 1999; 154 (1): 67-75.
  5. Rezuke W. N., Abernathy E. C., Tsongalis G. J. Molecular diagnosis of B- and T-cell lymphomas: fundamental principles and clinical applications. Clin. Chem. 1997; 43 (10): 1814- 1823.
  6. Arber D. A. Molecular diagnostic approach to non-Hodgkin'slymphoma. J. Mol. Diagn. 2000; 2 (4): 178-190.
  7. Arber D. A., Braziel R. M., Bagg A., Bijwaard К. Е. Evaluation of T cell receptor testing in limphoid neoplasms. Results of a Multicenter study of 29 extracted DNA and paraffin-embedded samples. Ibid. 2001; 3 (4): 133-140.
  8. Cabras A. D., Kremer M., Schulz S. et al. Quality assessment in diagnostic molecular pathology: experience from a German- Austrian-Swiss multicenter trial. Virchows Arch. 2000; 437 (1): 46-51.
  9. Greiner Т. С. Advances in molecular hematopathology: T-cell receptor у and bcl-2 genes. Am. J. Pathol. 1999; 154 (1): 7- 10.
  10. Mainl M. K., Casorati G., Dellabona P. et al. T-cell clonality in immune responses. Immunol. Today 1999; 20 (6): 262-266.
  11. Callan M. F., Steven N., Krausa P. et al. Large clonal expansions of CD8+ T cells in acute infectious mononucleosis. Nat. Med. 1996; 2 (8): 906-911.
  12. Southern E. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 1975; 98: 503-517.
  13. Langerok A. W., Wolvers-Tettero I. L. M., van Dongen J. J. M. Detection of T cell receptor beta (TCRB) gene rearrangement pattern in T-cell malignancies by Southern blot analysis. Leukemia 1999; 13: 965-974.
  14. Moreau E. J., Langerak A. W., van Gastel-Mol E. J. et al. Easy detection of all TCRG gene rearrangements by Southern blotanalysis: recommendations for optimal results. Ibid. 1620- 1626.
  15. Greiner T. C., Raffeld M., Lutz С. et al. Analysis of T-cell receptor-gamma gene rearrangements by denaturing gradient gelelectrophoresis of GC-clamped polymerase chain reaction products', correlation with tumor-specific sequences. Am. J. Pathol. 1995; 146: 46-55.
  16. Orita M., Iwahana H., Kanazawa H. et al. Detection of polymorphism by human DNA by gel electrophoresis as singlestrand conformation polymorphism. Proc. Natl. Acad. Sci. USA 1989; 86 (8): 209-214.
  17. Lynas C., Howe D. Simple, reliable detection of T cell clones by PCR-L1S-SSCP analysis of TCRy rearrangement. Mol. Cell. Probes 1998; 12: 41-48.
  18. Lefranc M.-P., Lefranc G. The T cell receptor facts book. London: Academic Press; 2001.
  19. Breit T. M., Wolvers-Tettero L. M., Hahlen K. et. al. Extensive junctional diversity of gamma delta T-cell receptors expressed by T-cell acute lymphoblastic leukemias: implications for the detection of minimal residual disease. Leukemia 1992; 6 (2): 169-170.
  20. Kneba M., Bolz I., Linke B. et al. Characterization of clonespecific rearrangement T-cell receptor gamma-chain genes in lymphomas and leukemias by the polymerase chain reaction and DNA sequencing. Blood 1994; 84 (2): 574-581.
  21. Theodorou I., Delfau Larue M. H., Bigorgne C. et al. Cutaneous T-cell infiltrates: analysis of T-cell receptor gamma gene rearrangement by polymerase chain reaction and denaturing gradient gel electrophoresis. Blood 1995; 86: 305-310.
  22. Dippel E., Assaf C., Hummel M. et al. Clonal T-cell receptor gamma-chain gene rearrangement by PCR-based GeneScan analysis in advanced cutaneous T-cell lymphoma: a critical evaluation. J. Pathol. 1999; 188: 146.
  23. Klemke С. D., Dippel E., Dembinski A. et al. Clonal T cell receptor gamma-chain gene rearrangement by PCR-based GeneScan analysis in the skin and blood of patients with parapsoriasis and early-stage mycosis fungoides. Ibid. 2002; 197 (3): 348-354.
  24. Theriault C., Galoin S., Valmary S. et al. PCR analysis of immunoglobulin Heavy chain (IgH) and TcR-gamma chain generearrangements in the diagnosis of lymphoproliferative disorders: Results of study of 525 cases. Mod. Pathol. 2000; 13 (12): 1269-1279.
  25. Miwa H., Nosaka Т., Kita K. et al. Immunogenotypes of lymphoid malignancies; the rearrangement of T cell receptor beta chain gene can occur before the gamma chain gene rearrangement. Jpn J. Cancer Res. 1988; 79 (4):484-490.
  26. Asou N., Matsuoka M., Hattori T. et al. T cell gamma gene rearrangements in hematologic neoplasms. Blood 1987; 69 (3): 968-970.
  27. Szczepanski Т., Langerak A. W., Wolvers-Tetero I. L. M. et al. Immunoglobulin and T cell receptor gene rearrangement pattern in acute lymphoblastic leukemia are less mature in adults than in children: implications for selection of PCR targets for detection of minimal residual disease. Leukemia 1998; 12: 1081-1088.
  28. Fodinger M., Winkler K., Mannhalter C., Chott A. Combined polymerase chain reaction approach for clonality detection in lymphoid neoplasms. Diagn. Mol. Pathol. 1999; 8 (2): 80-91.
  29. Wickham C. L., Lynas C., Ellard S. Detection of clonal T cell populations by high resolution PCR using fluorescently labelled nucleotides, evaluation using conventional LIS-SSCP. J. Clin. Mol. Pathol. 2000; 53: 150-154.
  30. Dippel E., Klemke D., Hummel M. et al. T-cell clonality of undetermined significance. Blood 2001; 98 (1): 247-248.
  31. Posnett D. N., Sinha R., Kabak S., Russo С. Clonal populations of T cells in normal elderly humans: the T cell equivalent to benign monoclonal gammapathy". J. Exp. Med. 1994; 179 (2): 609-618.

Copyright (c) 2003 Consilium Medicum

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Address of the Editorial Office:

  • Novij Zykovskij proezd, 3, 40, Moscow, 125167

Correspondence address:

  • Alabyan Street, 13/1, Moscow, 127055, Russian Federation

Managing Editor:

  • Tel.: +7 (926) 905-41-26
  • E-mail: e.gorbacheva@ter-arkhiv.ru


© 2018-2021 "Consilium Medicum" Publishing house

This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies