Cytokine-induced memory-like natural killer cells have enhanced function, proliferation, and in vivo expansion against ovarian cancer cells

• Published in: Gynecologic oncology 2019-04, Vol.153 (1), p.149-157
• Main Authors: Uppendahl, Locke D., Felices, Martin, Bendzick, Laura, Ryan, Caitlin, Kodal, Behiye, Hinderlie, Peter, Boylan, Kristin L.M., Skubitz, Amy P.N., Miller, Jeffrey S., Geller, Melissa A.
• Format: Article
• Language: English
• Subjects: Animals; Carcinoma, Ovarian Epithelial - immunology; Carcinoma, Ovarian Epithelial - pathology; Carcinoma, Ovarian Epithelial - therapy; Cell Line, Tumor; Cell Proliferation - drug effects; Cytokine-Induced Killer Cells - immunology; Cytokine-Induced Killer Cells - transplantation; Female; Humans; Immunologic Memory - immunology; Immunotherapy, Adoptive - methods; Interferon-gamma - biosynthesis; Interferon-gamma - immunology; Interleukin-12 - immunology; Interleukin-12 - pharmacology; Interleukin-15 - immunology; Interleukin-15 - pharmacology; Interleukin-18 - immunology; Interleukin-18 - pharmacology; Interleukins - immunology; Interleukins - pharmacology; Killer Cells, Natural - drug effects; Killer Cells, Natural - immunology; Lymphocyte Activation - drug effects; Lymphocyte Activation - immunology; Mice; Recombinant Proteins - immunology; Recombinant Proteins - pharmacology; Tumor Necrosis Factor-alpha - biosynthesis; Tumor Necrosis Factor-alpha - immunology; Xenograft Model Antitumor Assays
• ISSN: 0090-8258; 1095-6859
• DOI: 10.1016/j.ygyno.2019.01.006

Natural killer (NK) cells are lymphocytes well suited for adoptive immunotherapy. Attempts with adoptive NK cell immunotherapy against ovarian cancer have proven unsuccessful, with the main limitations including failure to expand and diminished effector function. We investigated if incubation of NK cells with interleukin (IL)-12, IL-15, and IL-18 for 16h could produce cytokine-induced memory-like (CIML) NK cells capable of enhanced function against ovarian cancer. NK cells were preactivated briefly with IL-12, IL-15, and IL-18, rested, then placed against ovarian cancer targets to assess phenotype and function via flow cytometry. Real-time NK-cell-mediated tumor-killing was evaluated. Using ascites cells and cell-free ascites fluid, NK cell proliferation and function within the immunosuppressive microenvironment was evaluated in vitro. Finally, CIML NK cells were injected intraperitoneal (IP) into an in vivo xenogeneic mouse model of ovarian cancer. CIML NK cells demonstrate enhanced cytokine (IFN-γ) production and NK-cell-mediated killing of ovarian cancer. NK cells treated overnight with cytokines led to robust activation characterized by temporal shedding of CD16, induction of CD25, and enhanced proliferation. CIML NK cells proliferate more with enhanced effector function compared to controls in an immunosuppressive microenvironment. Finally, human CIML NK cells exhibited potent antitumor effects within a xenogeneic mouse model of ovarian cancer. CIML NK cells have enhanced functionality and persistence against ovarian cancer in vitro and in vivo, even when exposed to ascites fluid. These findings provide a strategy for NK cell-based immunotherapy to circumvent the immunosuppressive nature of ovarian cancer. •Cytokine-induced memory-like (CIML) NK cells demonstrate enhanced function and proliferation against ovarian cancer cells.•A novel killing assay demonstrated enhanced CIML NK-cell-mediated cytolytic function against ovarian cancer in real time.•The functional and proliferative ability within immunosuppressive ascites was rescued with healthy donor CIML NK cells.•Human CIML NK cells exhibit potent antitumor effects within an in vivo mouse model of ovarian cancer.