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. 2018 Sep 3;215(9):2379–2395. doi: 10.1084/jem.20172168

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© 2018 Cichocki et al.

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Figure 1. — Adaptive NK cells exhibit heightened glycolysis and mitochondrial oxidative metabolism. (a) Metabolic functions in freshly isolated NK cells from five HCMV-seropositive and five HCMV-seronegative donors were analyzed by Seahorse. Shown are OCR profiles for representative donors and averages for maximal respiration, ATP-linked respiration, and SRC. (b) Quantification of ATP in NK cells from four seronegative and four seropositive donors. (c) ECAR profiles for representative donors and averages for maximal glycolysis and glycolytic reserve. (d) NK cells were isolated from five seronegative and five seropositive donors and incubated with the indicated inhibitors for 10 min before lysis and ATP quantification. Shown are fold decreases in ATP for each inhibitor relative to vehicle controls. (a–d) Results are from two independent experiments. (e) Associations between Seahorse metabolic readouts and the percentages of adaptive NK cells. Unpaired Student’s t tests were used to determine statistical significance. (f) Adaptive and canonical NK cells from three seropositive donors were sorted and analyzed by Seahorse. Shown is the OCR profile from a representative donor and averages for maximal respiration, ATP-linked respiration, and SRC. (g) Shown is the ECAR profile of canonical and adaptive NK cells from a representative donor, as well as the averages for maximal glycolysis and glycolytic reserve. All OCR and ECAR results are from two independent experiments. (h) Quantification of ATP in sorted canonical NK cells and adaptive NK cells from five seropositive donors. (i) Measurement of glucose uptake using 2-NBDG in isolated NK cells from six seropositive donors at rest, after stimulation with an anti-CD16 antibody and after stimulation with PMA:ionomycin. Shown are the 2-NBDG mean fluorescence intensity (MFI) averages in gated canonical and adaptive NK cells in each condition. Results are from two independent experiments. Paired Student’s t tests were used to determine statistical significance. Error bars represent SEM. *P < 0.05, **P < 0.01. n.s., not significant.