Cytokine Gene Expression in a Case of B-Cell Chronic Lymphocytic Leukemia (B-CLL) With an Unusual Expansion of T Cells at Presentation

  • Athanasia Mouzaki
  • Vincent Kindler
  • Nicolette Bowers
  • Arlette Doucet
  • Maria Melachrinou
  • Maria-Christina Kyrtsonis
  • Alice Kallinikou-Maniatis


We present a case of a 60 year old male with lymphadenopathy, hepatosplenomegaly and leucocytosis with lymphocytosis. At presentation, his WBC was 15.1x109/L with 75% lymphocytes; these were mainly T-cells with an atypical morphology, and a small monoclonal population of B cells, κ type. RT-PCR of immunoglobulin genes showed a monoclonal rearrangement of the IgM type. Elf-1, bcl-2, IL-1α and ß, IL-2, IL-4 and IL-6 genes were constitutively expressed to varying degrees, and superinduced upon mitogenic stimulation of the cells. Bandshifts with the lymphotropic transcription factors NF-kB, AP-1 and NF-AT, gave profiles typical of the T-cell leukemia Jurkat and not of normal (ex-vivo) T-cells or B-CLL cells, ex-vivo or stimulated. Bone marrow histology revealed a diffuse infiltration of lymphocytes, and immunohistochemistry on bone marrow sections the presence of both B and T-cells in almost equal proportions. At this stage a double (T/B) CLL was suspected. The patient was treated with alkylating agents and went into remission. Three months later, and while still on drugs, the patient’s WBC was found to be 5.43x109/L and immunophenotyping revealed a complete change of his T/B cell ratio. RT-PCR showed that the patient’s CD2 cells expressed none of the above genes but IL-4 when induced, and his CD19 cells IL-6 and IL-1ß when induced. The expreddion of IgM remained monoclonal, confirming the clinical observation of B leukemia development.


Chronic Lymphocytic Leukemia Electrophoretic Mobility Shift Assay Phorbol Myristate Acetate Peripheral Blood Leukocyte Phorbol Myristate Acetate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Matthes T, Werner-Favre C, Tang H, Zhang X, Kindler V, Zubler RH: Cytokine mRNA expression during in vitro response of human B lymphocytes: Kinetics of B-cell tumor necrosis factor a, interleukin (IL)-6, IL-10 and transforming growth factor b1 mRNAs. J Exp Med 178: 521, 1993PubMedCrossRefGoogle Scholar
  2. 2.
    Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chlorophorm extraction. Anal Biochem 162: 156, 1987PubMedCrossRefGoogle Scholar
  3. 3.
    Tang H, Matthes T, Carballido-Perrig, Zubler RH, Kindler V: Differential induction of T-cell cytokine mRNA in Epstein-Barr virus-transformed B-cell clones: Constitutive and inducible expression of interleukin-4 mRNA. Eur J Immunol 23: 899, 1993PubMedCrossRefGoogle Scholar
  4. 4.
    Trainor K, Brisco MJ, Story CJ, Morley AA: Monoclonality in B-lymphoproliferative disorders detected at the DNA level. Blood 75: 2220, 1990PubMedGoogle Scholar
  5. 5.
    Shi ZR, Itzkowitz SH, Kim YS: A comparison of three immunoperoxidase techniques for antigen detection in colorectal carcinoma tissues. J Histochem Cytochem 36: 317, 1988PubMedCrossRefGoogle Scholar
  6. 6.
    Poppema S, Hollema H, Visser L, Vos H: Monoclonal antibodies (MT1, MT2, MB1, MB2, MB3) reactive with leukocyte subsets in paraffin-embedded tissue sections. Am J Pathol 127: 418, 1987PubMedGoogle Scholar
  7. 7.
    Dobson CM, Myskow MW, Krajewski AS, Carpenter FH, Horne CHW: Immunohistochemical staining of non-Hodgkin’s lymphoma in paraffin sections using MB1 and MT1 monoclonal antibodies. J Pathol 153: 203, 1987PubMedCrossRefGoogle Scholar
  8. 8.
    Kuby J (ed.): Immunology, 2nd edition, W. H. Freeman and Co., New York, 1994, p 2971Google Scholar
  9. 9.
    Ehlers S, Smith KA: Differentiation of T-cell lymphokine gene expression: The in vitro acquisition of T-cell memory. J Exp Med 173: 25, 1991PubMedCrossRefGoogle Scholar
  10. 10.
    Mouzaki A, Rungger D: Properties of transcription factors regulating interleukin 2 gene transcription through the NF-AT binding site in untreated or drug-treated naive and memory T helper cells. Blood 84: 2612, 1994PubMedGoogle Scholar
  11. 11.
    Mouzaki A, Matthes T, Miescher PA, Beris Ph: Polyclonal hypergammaglobulinaemia in a case of a B-cell Chronic Lymphocytic leukaemia: The result of IL-2 production by the proliferating monoclonal B-cells? British J Haematol (in press)Google Scholar
  12. 12.
    Mouzaki A, Serfling E, Zubler RH: Interleukin-2 promoter activity in Epstein-Barr virus-transformed B lymphocytes is controlled by nuclear factor-kB. Eur J Immunol (in press)Google Scholar
  13. 13.
    Kishimoto T (ed): Interleukins: Molecular Biology and Immunology, in: Chemical Immunology, vol. 51, Karger, Basel, 1992Google Scholar
  14. 14.
    Fujita T, Shibuya H, Ohashi T, Yamanishi K, Taniguchi T: Regulation of interleukin-2 gene: functional DNA sequences in the 5’ flanking region for the gene expression in activated T lymphocytes. Cell 46: 403, 1986CrossRefGoogle Scholar
  15. 15.
    Shaw JP, Utz PJ, Durand DB, Toole JJ, Emmel EA, Crabtree GR: Identification of a putative regulator of early T-cell activation genes. Science 241: 202, 1988PubMedCrossRefGoogle Scholar
  16. 16.
    Kindler V, Matthes T, Jeannin P, Zubler RH: IL-2 secretion by human B lymphocytes occurs as a late event and requires additional stimulation after CD40 cross-linking. Eur J Immunol 25: 1239, 1995PubMedCrossRefGoogle Scholar
  17. 17.
    Rao A: NF-ATp: a transcription factor required for the co-ordinate induction of several cytokine genes. Immunology Today 15: 274, 1994PubMedCrossRefGoogle Scholar
  18. 18.
    Rooney JW, Hodge MR, McCaffrey PG, Rao A, Glimcher LH: A common factor regulates both Th1- and Th2-speciflc cytokine gene expression. EMBO J 13: 625, 1994PubMedGoogle Scholar
  19. 19.
    Yaseen NR, Maizel AL, Wang F, Sharma S: Comparative analysis of NF-AT (nuclear factor of activated T-cells) complex in human T and B lymphocytes. J Biol Chem 268: 14285, 1993PubMedGoogle Scholar
  20. 20.
    Thompson CB, Wang C-Y, Ho I-C, Bohjanen PR, Petryniak B, June CH, Miesfeldt S, Zhang L, Nabel GJ, Karpinski B, Leiden JM: cis-Acting sequences required for inducible interleukin-2 enhancer function bind a novel Ets-related protein, Elf-1. Mol Cel Biol 12: 1043, 1992Google Scholar
  21. 21.
    Liou H-C, Baltimore D: Regulation of the NF-kB/rel transcription factor and IkB inhibitor system. Current Opinion in Cell Biol 5: 477, 1993.CrossRefGoogle Scholar
  22. 22.
    Northorp JP, Ho SN, Chen l, Thomas DJ, Timmerman LA, Nolan GP, Admon A, Crabtree GR: NF-AT components define a family of transcription factors targeted in T-cell activation. Nature 369: 497, 1994CrossRefGoogle Scholar
  23. 23.
    Rai KR, Patel DV: Chronic lymphocytic leukemia, in Hoffman R, Benz EJ, Shattil SJ, Furie B, Cohen HJ, Silberstein LE (eds): Hematology, basic principles and practice, New York, Churchill Livingstone, 1995Google Scholar
  24. 24.
    John S, Reeves RB, Lin JX, Child R, Leiden JM, Thompson CB, Leonard WJ: Regulation of cell-type-specific interleukin-2 receptor alpha-chain gene expression: potential role of physical interactions between Elf-1, HMG-I(Y) and NF-kappa B family proteins. Mol Cell Biol 15: 1786, 199Google Scholar
  25. 25.
    Nolan GP: NF-AT-AP-1 and Rel-bZIP: Hybrid vigor and binding under the influence. Cell 77: 795, 1994PubMedCrossRefGoogle Scholar
  26. 26.
    Vaux DL, Cory S, Adams JM: Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B-cells. Nature 335: 440, 1988PubMedCrossRefGoogle Scholar
  27. 27.
    Thompson CB, Brown TA, McKnight SL: Convergence of ETS- and notch-related structural motifs in a heteromeric DNA binding complex. Science 253: 762, 1991PubMedCrossRefGoogle Scholar
  28. 28.
    Kang S-M, Beverley B, Tran A-C, Brorson K, Schwartz RH, Lenardo MJ: Transactivation by AP-1 is a molecular target of T-cell clonal anergy. Science 257: 1134, 1992PubMedCrossRefGoogle Scholar
  29. 29.
    Zaknoen SL, Christian SL, Suen R, Van Ness B, Kay NE: B-chronic lymphocytic leukemia cells contain both endogenous k immunoglobulin mRNA and critical immunoglobulin gene activation transcription factors. Leukemia 6: 675, 1992PubMedGoogle Scholar
  30. 30.
    Baltimore D, Beg AA: A butterfly flutters by. Nature 373: 287, 1995PubMedCrossRefGoogle Scholar
  31. 31.
    Hanada M, Delia D, Aiello A, Stadtmauer E, Reed JC: bcl-2 gene hypomethylation and high-level expression in B-cell chronic lymphocytic leukemia. Blood 6: 1820, 1993Google Scholar
  32. 32.
    Raziuddin S, Sheikha A, Abu-Eshy S, Al-Janadi M: Circulating levels of cytokines and soluble cytokine receptors in various T-cell malignancies. CANCER 73: 2426, 1994PubMedCrossRefGoogle Scholar
  33. 33.
    Rambaldi A, Bettoni S, Rossi V, Tini M-L, Giudici G, Rizzo V, Bassan R, Mantovani A, Barbui T, Biondi A: Transcripitonal and post-transcriptional regulation of IL-lb, IL-6 and TNF-a genes in chronic lymphocytic leukaemia. Br J Haematol 83: 204, 1993PubMedCrossRefGoogle Scholar
  34. 34.
    Mouzaki A, Rungger D: Interleukin 2 gene regulation at the Pud (NF-AT) promoter element: The complex situation in primary T lymphocytes. Molecular Biology of Hematopoiesis 3: 475, 1994Google Scholar
  35. 35.
    Harland CC, Whittaker SJ, NG YL, Holden CA, Wong E, Smith NP: Coexistent cutaneous T-cell lymphoma and B-cell chronic lymphocytic leukemia. Br J Dermatol 127: 519, 1992PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1996

Authors and Affiliations

  • Athanasia Mouzaki
    • 1
    • 2
    • 4
  • Vincent Kindler
    • 2
  • Nicolette Bowers
    • 2
  • Arlette Doucet
    • 2
  • Maria Melachrinou
    • 3
  • Maria-Christina Kyrtsonis
    • 1
  • Alice Kallinikou-Maniatis
    • 1
  1. 1.Hematology LaboratoryPatras University HospitalGreece
  2. 2.Division of HematologyGeneva University HospitalSwitzerland
  3. 3.Pathology LaboratoryPatras University HospitalGreece
  4. 4.Laboratory Hematology and Transfusion Medicine, Medical SchoolUniversity HospitalGreece

Personalised recommendations