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Optimization of cGMP purification and expansion of umbilical cord blood–derived T-regulatory cells in support of first-in-human clinical trials

  • David H. McKenna Jr.
    Correspondence
    Correspondence: David H. McKenna Jr., MD (mcken020@umn.edu), Clinical Cell Therapy Laboratory, University of Minnesota Medical Center, Molecular and Cellular Therapeutics, 1900 Fitch Avenue, St. Paul, MN 55108, USA.
    Affiliations
    Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Darin Sumstad
    Affiliations
    Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Diane M. Kadidlo
    Affiliations
    Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Bjorn Batdorf
    Affiliations
    Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Colin J. Lord
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Sarah C. Merkel
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Christine M. Koellner
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Julie M. Curtsinger
    Affiliations
    Department of Medicine, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Carl H. June
    Affiliations
    Department of Pathology and Laboratory Medicine, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA

    Abramson Family Cancer Center Research Institute, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA
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  • James L. Riley
    Affiliations
    Abramson Family Cancer Center Research Institute, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA

    Department of Microbiology, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA
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  • Bruce L. Levine
    Affiliations
    Department of Pathology and Laboratory Medicine, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA

    Abramson Family Cancer Center Research Institute, University of Pennsylvania Cancer Center, Philadelphia, Pennsylvania, USA
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  • Jeffrey S. Miller
    Affiliations
    Department of Medicine, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Claudio G. Brunstein
    Affiliations
    Department of Medicine, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • John E. Wagner
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Bruce R. Blazar
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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  • Keli L. Hippen
    Correspondence
    Correspondence: Keli L. Hippen, Ph.D, Department of Pediatrics, University of Minnesota, MMC366, 420 Delaware St. SE, Minneapolis, MN 55455, USA.
    Affiliations
    Department of Pediatrics, Hematology, Oncology and Transplantation, University of Minnesota, Minneapolis/Saint Paul, Minnesota, USA
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Published:November 22, 2016DOI:https://doi.org/10.1016/j.jcyt.2016.10.011

      Abstract

      Background aims

      Thymic-derived regulatory T cells (tTreg) are critical regulators of the immune system. Adoptive tTreg transfer is a curative therapy for murine models of autoimmunity, graft rejection, and graft-versus-host disease (GVHD). We previously completed a “first-in-human” clinical trial using in vitro expanded umbilical cord blood (UCB)-derived tTreg to prevent GVHD in patients undergoing UCB hematopoietic stem cell transplantation (HSCT). tTreg were safe and demonstrated clinical efficacy, but low yield prevented further dose escalation.

      Methods

      To optimize yield, we investigated the use of KT64/86 artificial antigen presenting cells (aAPCs) to expand tTreg and incorporated a single re-stimulation after day 12 in expansion culture.

      Results

      aAPCs increased UCB tTreg expansion greater than eightfold over CD3/28 stimulation. Re-stimulation with aAPCs increased UCB tTreg expansion an additional 20- to 30-fold. Re-stimulated human UCB tTreg ameliorated GVHD disease in a xenogeneic model. Following current Good Manufacturing Practice (cGMP) validation, a trial was conducted with tTreg. tTreg doses up to >30-fold higher compared with that obtained with anti-CD3/28 mAb coated-bead expansion and Foxp3 expression was stable during in vitro expansion and following transfer to patients. Increased expansion did not result in a senescent phenotype and GVHD was significantly reduced.

      Discussion

      Expansion culture with cGMP aAPCs and re-stimulation reproducibly generates sufficient numbers of UCB tTreg that exceeds the numbers of T effector cells in an UCB graft. The methodology supports future tTreg banking and is adaptable to tTreg expansion from HSC sources. Furthermore, because human leukocyte antigen matching is not required, allogeneic UCB tTreg may be a useful strategy for prevention of organ rejection and autoimmune disease.

      Key Words

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