Soluble and Exosome-Bound α-Galactosylceramide Mediate Preferential Proliferation of Educated NK Cells with Increased Anti-Tumor Capacity

Simple Summary Many cancers down-regulate the expression of major histocompatibility complex class I (MHC-I) to avoid recognition and elimination by the adaptive immune system. Natural Killer (NK) cells, as part of innate immunity, complement tumor recognition by their ability to sense foreign and malignant cells by the lack of major MHC-I expression. During development, NK cells learn to perceive physiological levels of self MHC-I as healthy in a process termed NK cell education. In the current study, we assessed whether the stimulation of natural killer T (NKT) cells could impact the tumor-killing ability of NK cells. We can show that stimulation with a well-established activator of NKT cells induces the preferential expansion of educated NK cells, and improves NK-cell-mediated tumor-killing of both MHC-I+ and MHC-I− tumor cells. Furthermore, this increased tumor-killing capacity of NK cells can be achieved by nanovesicle-mediated delivery, which is known to improve adaptive immunity as well. Our approach thus holds great potential for future cancer immunotherapy. Abstract Natural killer (NK) cells can kill target cells via the recognition of stress molecules and down-regulation of major histocompatibility complex class I (MHC-I). Some NK cells are educated to recognize and kill cells that have lost their MHC-I expression, e.g., tumor or virus-infected cells. A desired property of cancer immunotherapy is, therefore, to activate educated NK cells during anti-tumor responses in vivo. We here analyze NK cell responses to α-galactosylceramide (αGC), a potent activator of invariant NKT (iNKT) cells, or to exosomes loaded with αGC. In mouse strains which express different MHC-I alleles using an extended NK cell flow cytometry panel, we show that αGC induces a biased NK cell proliferation of educated NK cells. Importantly, iNKT cell-induced activation of NK cells selectively increased in vivo missing self-responses, leading to more effective rejection of tumor cells. Exosomes from antigen-presenting cells are attractive anti-cancer therapy tools as they may induce both innate and adaptive immune responses, thereby addressing the hurdle of tumor heterogeneity. Adding αGC to antigen-loaded dendritic-cell-derived exosomes also led to an increase in missing self-responses in addition to boosted T and B cell responses. This study manifests αGC as an attractive adjuvant in cancer immunotherapy, as it increases the functional capacity of educated NK cells and enhances the innate, missing self-based antitumor response.