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. Author manuscript; available in PMC: 2016 Feb 15.
Published in final edited form as: J Immunol. 2015 Jan 16;194(4):1954–1962. doi: 10.4049/jimmunol.1402099

Figure 6. Exposure to high-dose IL-15, but not proliferation, alters the metabolic requirements for receptor-stimulated IFN-γ production.

Figure 6

(A) NK cells were incubated for 14–16hr with IL-15 (10 or 100ng/ml), IL-12, and/or IL-18, washed, and then stimulated for 6h with anti-NK1.1 or control IgG in the presence or absence of oligomycin (Oligo). Results represent the mean +/−SEM from 4 independent experiments; statistics represent comparison of anti-NK1.1 versus anti-NK1.1 plus oligomycin. (B&C) NK cells were cultured for 72hr with 100ng/ml IL-15, washed, and activated (6hr) with cytokines, PMA + calcimycin (PMA+CA), or receptors (anti-NK1.1, anti-Ly49D or control IgG) in the presence of (B) oligomycin (100nM) or (C) glucose-free (GF) media, glutamine-free media, etomoxir, or glutamine-free media. Results represent the mean +/−SEM from 3 independent experiments. (D) NK cells were cultured for 72h with 10ng/ml or 100ng/ml IL-15 followed by anti-NK1.1 stimulation in the presence or absence of oligomycin. (E) Production of IFN-γ by highly divided and undivided NK cells, as measured by CFSE dilution following IL-15 activation (100ng/ml). Results for D&E are representative of 3 independent experiments. (F) Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of NK cells treated with IL-15 (100ng/ml) for 72h compared to freshly isolated NK cells rested in media for 4hr. Results represent the mean +/−SEM of 3–6 replicate wells from 2 independent experiments. *p≤0.05; **p≤0.01, ***p≤0.001.