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International Journal of Hematology

, Volume 108, Issue 6, pp 588–597 | Cite as

Dasatinib-induced anti-leukemia cellular immunity through a novel subset of CD57 positive helper/cytotoxic CD4 T cells in chronic myelogenous leukemia patients

  • Naoki Watanabe
  • Tomoiku Takaku
  • Kazuyoshi Takeda
  • Shuichi Shirane
  • Tokuko Toyota
  • Michiaki Koike
  • Masaaki Noguchi
  • Takao Hirano
  • Hiroshi Fujiwara
  • Norio Komatsu
Original Article

Abstract

Dasatinib induces lymphocytosis of large granular lymphocytes (LGLs) in a proportion of patients with chronic myelogenous leukemia (CML), and is associated with better clinical outcomes. LGLs consist of cytotoxic T lymphocytes and natural killer cells; however, the context and phenotypic/functional features of each type of LGL are unknown. To better define features of these LGLs, we investigated lymphocytosis in CML patients treated with dasatinib. D57-positive and CD4-positive type I T-helper (Th) cells (CD57+ Th cells) rarely occur in CML patients without lymphocytosis and in healthy individuals; however, a substantial increase in the proportion of CD57+ Th cells was observed in CML patients treated with dasatinib. In addition, these cells showed appreciable levels of cytocidal activity via cytotoxic degranulation. Analysis of T-cell receptor α and β sequences showed a skewed T-cell repertoire in the CD57+ Th cells. Furthermore, patients with LGLs and CD57+ Th lymphocytosis achieved stronger molecular responses than did those without lymphocytosis. While further studies are warranted, our observations suggest that dasatinib induces the expansion of CD57+ Th-LGLs, which may play a crucial role in the dasatinib-induced response against Philadelphia chromosome-positive leukemia.

Keywords

CD57 expression Chronic myelogenous leukemia Cytotoxic CD4+ T cell Dasatinib Large granular lymphocyte 

Notes

Acknowledgements

We are grateful to Ryuji Suzuki and Kazutaka Kitaura from Repertoire Genesis Incorporated, for providing technical support.

Author contributions

NW performed the study, analyzed the results, and wrote the paper; TTa designed the research, analyzed the results, wrote the paper, and directed the research; KT and SS analyzed the results; TTo performed the study; MK, MN, and TH contributed analytical materials; HF discussed and analyzed data, provided experimental concepts and materials, and edited the paper; and NK directed the research.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest. A summary of relevant information will be published with the manuscript.

Supplementary material

12185_2018_2517_MOESM1_ESM.tif (24.9 mb)
Supplemental Figure 1 Representative immunophenotyping analysis of large granular lymphocytes (LGLs). CD56+ natural killer (NK) cells in patients (a) without and (b) with LGL lymphocytosis. CD57+ NK cells in patients (c) without and with (d) LGL lymphocytosis. CD57+ cytotoxic T lymphocytes in patients (e) without and (f) with LGL lymphocytosis. CD57+ helper T cells in patients (g) without and (h) with LGL lymphocytosis (TIF 25509 KB)
12185_2018_2517_MOESM2_ESM.tif (24.9 mb)
Supplemental Figure 2 Comparison of cytoplasmic granules in CD57-positive and -negative Th cells. Cytoplasmic granules by Giemsa staining indicated morphologic features of sorted CD57 positive and negative helper T (Th) cells. (a) CD57+ Th cells showed cytoplasmic granules. (b) CD57 negative Th cells did not show cytoplasmic granules. (TIF 25507 KB)
12185_2018_2517_MOESM3_ESM.tif (27.2 mb)
Supplemental Figure 3 Immunophenotyping analysis of CD57+ Th cells. Flow cytometric analysis of representative T cells. (a) Expression of CD45RA and CCR7. (b) interferon (IFN)-γ and interleukin (IL)-4. (c) IFN-γ and IL-17 in CD57+ Th cells. T-cell receptor (TCR) Vα24-Jα18 expression in (d) CD57+ Th cells, (e) CD3+ CD4- CD57+ cells, and in (f) CD3+ CD4− CD57− cells. (TIF 27896 KB)
12185_2018_2517_MOESM4_ESM.tif (24.9 mb)
Supplemental Figure 4 Analysis of intracellular cytokine production without stimulation in CD57+ helper T (Th) cells. CD57+ Th cells without phorbol 12-myristate 13-acetate (PMA) and ionomycin (ION) stimulation did not produce any cytokine. (a) interferon (IFN)-γ and interleukin (IL)-4. (b) IFN-γ and IL-17. For intracellular cytokine staining, the sorted effector cells were cultured with PMA (50 ng/mL) and ION (500 ng/mL) (Sigma-Aldrich) for 12 h, with the addition of Brefeldin A (Sigma-Aldrich) 2 h before the end of incubation. The cells were then fixed with 4% paraformaldehyde, permeabilized with permeabilization buffer (eBioscience), and stained with antibodies. (TIF 25508 KB)
12185_2018_2517_MOESM5_ESM.tif (24.9 mb)
Supplemental Figure 5 Cytotoxicity of CD57+/− helper T (Th) cells targeting K562-A24/C II TA and K562-A24. The solid line represents the cytotoxicity of CD3+ CD4+ CD57+ Th cells and the broken line represents the cytotoxicity of CD3+ CD4+ CD57− Th cells. (TIF 25507 KB)

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Copyright information

© The Japanese Society of Hematology 2018

Authors and Affiliations

  • Naoki Watanabe
    • 1
  • Tomoiku Takaku
    • 1
  • Kazuyoshi Takeda
    • 2
    • 3
  • Shuichi Shirane
    • 1
  • Tokuko Toyota
    • 1
  • Michiaki Koike
    • 4
  • Masaaki Noguchi
    • 5
  • Takao Hirano
    • 6
  • Hiroshi Fujiwara
    • 7
  • Norio Komatsu
    • 1
  1. 1.Department of HematologyJuntendo University School of MedicineTokyoJapan
  2. 2.Division of Cell Biology, Biomedical Research Center, Graduate School of MedicineJuntendo UniversityTokyoJapan
  3. 3.Department of Biofunctional Microbiota, Graduate School of MedicineJuntendo UniversityTokyoJapan
  4. 4.Department of HematologyJuntendo Shizuoka HospitalShizuokaJapan
  5. 5.Department of HematologyJuntendo Urayasu HospitalChibaJapan
  6. 6.Department of HematologyJuntendo Nerima HospitalTokyoJapan
  7. 7.Department of Hematology, Clinical Immunology and Infectious Diseases, Ehime University Graduate School of MedicineEhime UniversityToonJapan

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