Preview

Russian journal of hematology and transfusiology

Advanced search

FLOW CYTOMETRY AND PCR-BASED T-CELL CLONALITY TESTING FOR DISCRIMINATING BETWEEN REACTIVE AND NEOPLASTIC PROLIFERATION OF LARGE GRANULAR LYMPHOCY

https://doi.org/10.25837/HAT.2018.48..2..003

Abstract

Background. Proliferation of large granular lymphocytes (LGL) can be associated with viral infections, autoimmune disorders, chronic lymphoproliferative disorder of NK-cells (CLPD-NK), or T-cell large granular lymphocytic leukemia (T-LGL). Differential diagnosis between reactive and neoplastic LGL proliferation could be a challenge.
Objective. To evaluate the role of morphological, immunophenotypic (flow cytometry) and molecular genetic (PCR-based T-cell clonality) studies in differential diagnosis of LGL proliferation.
Materials and methods. 68 patients with increased peripheral blood LGL counts were enrolled in the study. Male/female ratio was 20/48; the median age was 53 (22—79) years. Blood smear, immunophenotyping by flow cytometry, and PCR-based T-cell clonality assays (TCRG (V γ-J γ) and TCRG (D β-J β and V β-J β)) were performed in all cases.
Results. We found that morphology, immunophenotype, relative and absolute LGL counts, or heterogenic expression of T-cell markers can not distinguish between neoplastic and reactive LGL proliferation. Clonal TCRG (V γ-J γ) and TCRG (D β-J β) gene rearrangements were observed in reactive LGL lymphocytes and T-LGL. Complete rearrangements of TCRG (V β-J β) genes were specific for malignancies and were not detected in any case of reactive lymphocytosis.

About the Authors

N. G. Chernova
National Research Center for Hematology
Russian Federation
Chernova Natalia, MD, PhD, hematologist in National Research Center for Hematology, Moscow, 125167


Yu. V. Sidorova
National Research Center for Hematology, Moscow
Russian Federation


E. I. Zakharko
National Research Center for Hematology, Moscow
Russian Federation


E. V. Naumova
Department of clinical laboratory diagnostics, Russian Medical Academy of Continuous Professional Education, Moscow
Russian Federation


N. V. Ryzhikova
National Research Center for Hematology, Moscow
Russian Federation


I. V. Galtseva
National Research Center for Hematology, Moscow
Russian Federation


V. N. Dvirnyk
National Research Center for Hematology, Moscow
Russian Federation


S. Yu. Smirnova
National Research Center for Hematology, Moscow
Russian Federation


A. B. Sudarikov
National Research Center for Hematology, Moscow
Russian Federation


A. V. Kokhno
National Research Center for Hematology, Moscow
Russian Federation


T. N. Moiseeva
National Research Center for Hematology, Moscow
Russian Federation


S. A. Lugovskaya
Department of clinical laboratory diagnostics, Russian Medical Academy of Continuous Professional Education, Moscow
Russian Federation


E. E. Zvonkov
National Research Center for Hematology, Moscow
Russian Federation


V. G. Savchenko
National Research Center for Hematology, Moscow
Russian Federation


References

1. Loughran T. P. Jr. Clonal diseases of large granular lymphocytes. Blood. 1993; 82(1):1—14.

2. Sokol L., Loughran T. P. Jr. Large granular lymphocyte leukemia. Oncologist. 2006; 11(3):263—273. DOI: 10.1007/s11899-007-0038-7.

3. Lamy T., Loughran T. P. Jr. Large granular lymphocyte leukemia. Cancer Control. 1998; 5(1):25—33.

4. Swerdlow S. H., Campo E., Harris N. L. et al. WHO classification of tumors of haematopoietic and lymphoid tissues. Lyon: IARC; 2017, 348—350.

5. Zambello R., Trentin L., Facco M. et al. Analysis of the T cell receptor in the lymphoproliferative disease of granular lymphocytes: superantigen activation of clonal CD3+ granular lymphocytes. Cancer Res. 1995; 55(24):6140—6145.

6. Vinogradova Yu. E., Lutsenko I. N., Kremenetskaya A. M. et al. Structure of T/NK-cell lymphomas in the National Research Center for Hematology of the Russian Academy of Medical Sciences. Russian Journal of Problems of Hematology and Blood Transfusion (Problemy gematologii i perelivaniya krovi). 2005; 50(4):30—34. (In Russian)

7. Lamy T., Loughran T. P. Jr. Clinical features of large granular lymphocyte leukemia. Semin Hematol. 2003; 40(3):185—195.

8. Sampalo Lainz A., López-Gómez M., Jimenez-Alonso J. Proliferation of large granular lymphocytes in patients with systemic lupus erythematosus. Rev Clin Esp. 1995; 195(6):373—379.

9. Starkebaum G. Chronic neutropenia associated with autoimmune disease. Semin Hematol. 2002; 39(2):121—127.

10. Morice W. G., Kurtin P. J., Leibson P. J. et al. Demonstration of aberrant T-cell and natural killer-cell antigen expression in all cases of granular lymphocytic leukaemia. Br J Haematol 2003; 120(6):1026—1036.

11. Semenzato G., Zambello R., Starkebaum G. et al. The lymphoproliferative disease of granular lymphocytes: updated criteria for diagnosis. Blood. 1997; 89(1):256—260.

12. Vinogradova Yu. E., Lutsenko I. N., Samoylova R. S. et al. Clinical and immunomorphological variants of large granular lymphocyte T/NK lymphocytic leukemia. Russian Journal of Hematology and Transfusiology (Gematologiya i transfuziologiya). 2009; 54(2):14—18. (In Russian)

13. Vie H., Chevalier S., Garand R. et al. Clonal expansion of lymphocytes bearing the gamma delta T-cell receptor in a patient with large granular lymphocyte disorder. Blood. 1989; 74(1):285—290.

14. Dongen J. J., Langerak A. W., Bruggemann M. et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia. 2003; 17(12):2257—2317. DOI: 10.1038/sj.leu.2403202.

15. Khaydukov S. V., Baydun L. V. Modern approach to assessing cellular constituents immune status. Russian Journal of Medical Alphabet. (Meditsinskiy alfavit). 2015; 2(8):44—51. (In Russian)

16. Vorobiev A. I., Vorobiev I. A., Gretsov E. M. et al. Problems of the theoretical hematology. Russian Journal of Therapeutic Archive (Terapevticheskiy arkhiv). 2003; 75(9):22—29. (In Russian)

17. O’Keefe C. L., Plasilova M., Wlodarski M. et al. Molecular analysis of TCR clonotypes in LGL:a clonal model for polyclonal responses. J Immunol. 2004; 172(3):1960—1969.

18. Wlodarski M. W., O’Keefe C., Howe E. C. et al. Pathologic clonal cytotoxic T-cell responses: nonrandom nature of the T-cell-receptor restriction in large granular lymphocyte leukemia. Blood. 2005; 106(8):2769—2780. DOI: 10.1182/blood-2004-10-4045.

19. Pandolfi F., Loughran T. P. Jr., Starkebaum G. et al. Clinical course and prognosis of the lymphoproliferative disease of granular lymphocytes. A multicenter study. Cancer. 1990; 65(2):341—348.

20. Sidorova Yu. V., Chernova N. G., Ryzhikova N. V. et al. Clonal rearrangements and malignant clones in peripheral T-cell lymphoma. Acta Naturae (English version). 2015; 7(3):116—125.

21. Smirnova S., Sidorova Y., Chernova N. et al. CD8+ T-cell clones persistent in bone marrow and peripheral blood during course of CD4+ angioimmunoblastic T-cell lymphoma. Haematologica. 2017; 102(1):578, E1403.

22. Sidorova Yu. V., Nikitin E. A., Melikyan A. L. et al. Use of PCR-SSCPTCR γ method to determine T-cell clonality in infectious mononucleosis. Russian Journal of Hematology and Transfusiology (Gematologiya i transfuziologiya). 2004; 49(6):1—7. (In Russian)

23. Smirnova S. J., Sidorova J. V., Biderman B. V. et al. Expansion of CD8+ cells in autoimmune hemolytic anemia. Autoimmunity. 2 016; 49(3):147— 154. DOI: 10.3109/08916934.2016.1138219.

24. French L.E., Alcindor T., Shapiro M. et al. Identification of amplified clonal T cell populations in the blood of patients with chronic graftversus-host disease:positive correlation with response to photopheresis. Bone Marrow Transplant. 2002; 30(8):509—515. DOI: 10.1038/sj.bmt.1703705.

25. Narumi H., Kojima K., Matsuo Y. et al. T-cell large granular lymphocytic leukemia occurring after autologous peripheral blood stem cell transplantation. Bone Marrow Transplant. 2004; 33(1):99—101. DOI: 10.1038/sj.bmt.1704298.

26. Au W. Y., Lam C. C., Lie A. K. et al. T-cell large granular lymphocyte leukemia of donor origin after allogeneic bone marrow transplantation. Am J Clin Pathol 2003; 120(4):626—630. DOI: 10.1309/VA75-5A03-PVRV-9XDT.

27. Sychevskaya K. A., Smirnova S., Sidorova J. et al. Clonal T-lymphocyte populations in healthy donors. Blood. 2017; 130(1):3598.


Review

For citations:


Chernova N.G., Sidorova Yu.V., Zakharko E.I., Naumova E.V., Ryzhikova N.V., Galtseva I.V., Dvirnyk V.N., Smirnova S.Yu., Sudarikov A.B., Kokhno A.V., Moiseeva T.N., Lugovskaya S.A., Zvonkov E.E., Savchenko V.G. FLOW CYTOMETRY AND PCR-BASED T-CELL CLONALITY TESTING FOR DISCRIMINATING BETWEEN REACTIVE AND NEOPLASTIC PROLIFERATION OF LARGE GRANULAR LYMPHOCY. Russian journal of hematology and transfusiology. 2018;63(2):124-133. (In Russ.) https://doi.org/10.25837/HAT.2018.48..2..003

Views: 2231


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 0234-5730 (Print)
ISSN 2411-3042 (Online)