Good manufacturing practices production of natural killer cells for immunotherapy: a six-year single-institution experience

Authors

  • David H. McKenna Jr.,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Darin Sumstad,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Nancy Bostrom,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Diane M. Kadidlo,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Susan Fautsch,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Sarah McNearney,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Rose DeWaard,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Philip B. McGlave,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Daniel J. Weisdorf,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • John E. Wagner,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Jeffrey McCullough,

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author
  • Jeffrey S. Miller

    1. From the Department of Laboratory Medicine and Pathology, Division of Transfusion Medicine, University of Minnesota Medical School; the Clinical Cell Therapy Laboratory, University of Minnesota Medical Center; and the Departments of Medicine and Pediatrics, Division of Hematology, Oncology, and Transplantation, University of Minnesota Cancer Center, Minneapolis/St Paul, Minnesota.
    Search for more papers by this author

  • Supported in part by the National Heart, Lung, and Blood Institute Contract N01-HB-47095 (Production Assistance for Cellular Therapies, or PACT), P01 CA111412 (DJW, JSM), P01 65493 (PBM, JSM, JEW, DHM), M01-RR00400 (National Center for Research Resources), and the BMT Research Fund and Children's Cancer Research Fund (JEW, JSM).

David H. McKenna, Jr, MD, Clinical Cell Therapy Laboratory, University of Minnesota Medical Center, Molecular & Cellular Therapeutics, 1900 Fitch Avenue, St Paul, MN 55108; e-mail: mcken020@umn.edu.

Abstract

BACKGROUND: Natural killer (NK) cells, a subset of lymphocytes and part of the innate immune system, play a crucial role in defense against cancer and viral infection. Herein is a report on the experience of clinical-scale, good manufacturing practices (GMPs) production of NK cells to treat advanced cancer.

STUDY DESIGN AND METHODS: Two types of NK cell enrichments were performed on nonmobilized peripheral blood mononuclear cell apheresis collections with a cell selection system (CliniMACS, Miltenyi): CD3 cell depletion to enrich for NK cells and CD3 cell depletion followed by CD56 cell selection to obtain a more pure NK cell product. After overnight incubation with interleukin-2 (IL-2), cells were washed, resuspended in 5 percent human serum albumin, and then released for infusion.

RESULTS: A total of 70 NK cell therapy products have been manufactured for patient infusion since 2000. For the CD3 cell–depleted NK cell products, the mean purity, recovery, and viability were 38, 79, and 86 percent, respectively. For the CD3 cell–depleted/CD56 cell–enriched NK cell products, the mean purity, recovery, and viability were 90, 19, and 85 percent, respectively. Gram stain, sterility, and endotoxin testing were all within acceptable limits for established lot release. Compared to the resting processed cells, IL-2 activation significantly increased the function of cells in cytotoxicity assays.

CONCLUSION: Clinical-scale production of NK cells is efficient and can be performed under GMPs. The purified NK cell product results in high NK cell purity with minimal contamination by T cells, monocytes, and B cells, but it requires more time for processing and results in a lower NK cell recovery when compared to NK cell enrichment with CD3 cell depletion alone. Additional laboratory studies and results from clinical trials will identify the best source and type of NK cell product.

Ancillary