IL-12 Directs Further Maturation of Ex Vivo Differentiated NK Cells with Improved Therapeutic Potential

Abstract

The possibility to modulate ex vivo human NK cell differentiation towards specific phenotypes will contribute to a better understanding of NK cell differentiation and facilitate tailored production of NK cells for immunotherapy. In this study, we show that addition of a specific low dose of IL-12 to an ex vivo NK cell differentiation system from cord blood CD34+ stem cells will result in significantly increased proportions of cells with expression of CD62L as well as KIRs and CD16 which are preferentially expressed on mature CD56dim peripheral blood NK cells. In addition, the cells displayed decreased expression of receptors such as CCR6 and CXCR3, which are typically expressed to a lower extent by CD56dim than CD56bright peripheral blood NK cells. The increased number of CD62L and KIR positive cells prevailed in a population of CD33+NKG2A+ NK cells, supporting that maturation occurs via this subtype. Among a series of transcription factors tested we found Gata3 and TOX to be significantly downregulated, whereas ID3 was upregulated in the IL-12-modulated ex vivo NK cells, implicating these factors in the observed changes. Importantly, the cells differentiated in the presence of IL-12 showed enhanced cytokine production and cytolytic activity against MHC class I negative and positive targets. Moreover, in line with the enhanced CD16 expression, these cells exhibited improved antibody-dependent cellular cytotoxicity for B-cell leukemia target cells in the presence of the clinically applied antibody rituximab. Altogether, these data provide evidence that IL-12 directs human ex vivo NK cell differentiation towards more mature NK cells with improved properties for potential cancer therapies.

Bortezomib augments lymphocyte stimulatory cytokine signaling in the tumor microenvironment to sustain CD8+T cell antitumor function

“…The effects of bortezomib treatment on these cytokines, which are key players in the cytotoxic and memory response mediated by CD8 + T cells and NK cells [36–38], suggest that bortezomib has the potential to influence the tumor microenvironment and host antitumor immunity.…”

Combined IL-15 and IL-12 drives the generation of CD34+-derived natural killer cells with superior maturation and alloreactivity potential following adoptive transfer

“…29 In the present study, we further investigated the influence of IL-12 on ex vivo-generated HPC-NK cells in vitro and in vivo. We demonstrated that replacement of IL-2 by IL-12 favors the development of KIRexpressing NK cells, which display superior IFNg production and cytolytic activity against AML cells, including cell lines and patient-derived material.…”

“…However, several other cytokines are known to promote NK cell development, activation, and function. [27][28][29] Although IL-15 plays a crucial role during NK cell development as well as in the survival and expansion of NK cells in vivo, recent studies also demonstrated that preactivation with combinations of the cytokines IL-12, IL-15, and IL-18 results in "priming" of memory-like NK cells with enhanced functional properties. [30][31][32] Although the precise underlying mechanism remains unknown, this prime-boost strategy using distinct combinations of cytokines may pave the way to improved NK cell immunotherapeutic strategies.…”

Oncolytic Virus Encoding a Master Pro-Inflammatory Cytokine Interleukin 12 in Cancer Immunotherapy

“…Interleukin 12 (IL-12) is a powerful master regulator of both innate and adaptive anti-tumor immune responses. As a heterodimeric cytokine, it produces multifaceted anti-tumor effects [1,2], including stimulation of growth and cytotoxic activity of natural killer (NK) cells and T cells (both CD4 + and CD8 + ) [1,[3][4][5], induction of differentiation of CD4 + T cells towards Th1 phenotype [6,7], increased production of IFN-γ from NK and T cells [1,8,9], and inhibition of tumor angiogenesis [1,10]. Despite encouraging success in preclinical studies [4], the early stages of IL-12 clinical trials did not meet expectations.…”