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Experimental Therapy

Diphtheria toxin fused to human interleukin-3 is toxic to blasts from patients with myeloid leukemias

Leukemia volume 14, pages 576–585 (2000)Cite this article

Abstract

Leukemic blasts from patients with acute phase chronic myeloid leukemic and refractory acute myeloid leukemia are highly resistant to a number of cytotoxic drugs. To overcome multi-drug resistance, we engineered a diphtheria fusion protein by fusing human interleukin-3 (IL3) to a truncated form of diphtheria toxin (DT) with a (G4S)2 linker (L), expressed and purified the recombinant protein, and tested the cytotoxicity of the DTLIL3 molecule on human leukemias and normal progenitors. The DTLIL3 construct was more cytotoxic to interleukin-3 receptor (IL3R) bearing human myeloid leukemia cell lines than receptor-negative cell lines based on assays of cytotoxicity using thymidine incorporation, growth in semi-solid medium and induction of apoptosis. Exposure of mononuclear cells to 680 pM DTLIL3 for 48 h in culture reduced the number of cells capable of forming colonies in semi-solid medium (colony-forming units leukemia) ⩾10-fold in 4/11 (36%) patients with myeloid acute phase chronic myeloid leukemia (CML) and 3/9 (33%) patients with acute myeloid leukemia (AML). Normal myeloid progenitors (colony-forming unit granulocyte–macrophage) from five different donors treated and assayed under identical conditions showed intermediate sensitivity with three- to five-fold reductions in colonies. The sensitivity to DTLIL3 of leukemic progenitors from a number of acute phase CML patients suggests that this agent could have therapeutic potential for some patients with this disease.

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Acknowledgements

This work was supported in part by Leukemia Society of America Grant No. 6114–98 (to AF), NIH Grants No. R01CA76178 (to AF), R01CA54116 (to ET), R01CA51025 (to JM) and grants from the National Cancer Institutes of Canada (NCIC) to DH with funds from the Terry Fox Run.

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Authors and Affiliations

  1. Department of Cancer Biology, Wake Forest University School of Medicine, Winston-Salem, NC, USA

    AE Frankel, MS Miller, J Ramage, M Kiser, GL Kucera & RL Alexander

  2. Department of Microbiology, East Carolina University, Greenville, NC, USA

    JA McCubrey

  3. Departments of Surgery, Medical University of South Carolina, Charleston, SC, USA

    S Delatte & EP Tagge

  4. Department of Medicine, MD Anderson Cancer Center, Houston, TX, USA

    M Beran

  5. Laboratory of Molecular Biology, National Cancer Institute, NIH, Bethesda, MD, USA

    RJ Kreitman

  6. Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada

    DE Hogge

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Frankel, A., McCubrey, J., Miller, M. et al. Diphtheria toxin fused to human interleukin-3 is toxic to blasts from patients with myeloid leukemias. Leukemia 14, 576–585 (2000). https://doi.org/10.1038/sj.leu.2401743

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