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Anti-thymocyte globulin’s activity against acute myeloid leukemia stem cells

Bone Marrow Transplantation (2018) | Download Citation

Abstract

Rabbit anti-thymocyte globulin (ATG (Thymoglobulin)) kills T cells in vitro and probably also in vivo as it prevents graft-vs-host disease (GvHD) in patients. Recently we demonstrated that ATG at a clinically relevant concentration (10–50 mg/L) kills in vitro not only T cells but also leukemic blasts. In the present study, we investigated whether ATG kills not only leukemic blasts but also leukemic stem cells (LSCs). We used a flow cytometric assay of complement-mediated cytotoxicity (CDC). ATG-induced death of acute myeloid leukemia (AML) cells from patients newly diagnosed with AML was measured among blasts as well as LSCs. At 10 mg/L ATG, blasts but not LSCs were killed. At 50 mg/L ATG, both blasts and LSCs were killed. We also measured ATG-mediated killing of healthy individuals’ hematopoietic stem cells (HSCs). Median 2% HSCs from blood and 15% HSCs from filgrastim-mobilized grafts were killed with 50 mg/L ATG, compared to 30% LSCs from the blood of AML patients (p = 0.001 and 0.022, respectively). In conclusion, LSCs are sensitive to ATG, however, only at a relatively high ATG concentration. At that concentration, LSCs are killed to a higher degree than HSCs.

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Acknowledgements

We thank the patients for participating in research that could not benefit them but only future patients. We thank the healthy volunteers involved in the study. This study could not happen without the dedication of Mamta Kantharia, Jennifer LeBlanc, Lori Rackel, Laura Spilchen, many inpatients nurses, pharmacists, particularly Michelle Dowhan, and physicians, notably Dr. Michelle Geddes, Dr. Mona Shafey, Dr. Peter Duggan, and Dr. Lynne Savoie. We also thank the staff of Calgary Laboratory Services, including Glenis Doiron. Finally, we thank Douglas Mahoney for invaluable feedback during this study.

Author contributions

R.D. developed the cytotoxicity assays, ran the assays, performed statistical analysis, and analyzed the results. P.D.K. and J.L. provided input into assay development and interpretation. M.M., T.v.S., and J.B. collected or arranged for the collection of specimens. A.D., D.M., F.M.K., and J.S. provided critical feedback. R.D. and J.S. designed the study, and J.S. supervised its conduct. R.D. wrote the manuscript.

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Affiliations

  1. University of Calgary, Calgary, AB, Canada

    • Rosy Dabas
    • , Poonam Dharmani-Khan
    • , Monica Modi
    • , Don Morris
    • , Andrew Daly
    • , Faisal M. Khan
    •  & Jan Storek
  2. Alberta Health Services, Calgary, AB, Canada

    • Poonam Dharmani-Khan
    • , Monica Modi
    • , Tiffany Van Slyke
    • , Joanne Luider
    • , Don Morris
    • , Joseph Brandwein
    • , Andrew Daly
    • , Faisal M. Khan
    •  & Jan Storek
  3. University of Alberta, Edmonton, AB, Canada

    • Tiffany Van Slyke
    •  & Joseph Brandwein

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The authors declare that they have no conflict of interest.

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Correspondence to Rosy Dabas.

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https://doi.org/10.1038/s41409-018-0296-0