Extended Data Fig. 6: Impact of acylcarnitines on immune cell function.
From: The source of dietary fat influences anti-tumour immunity in obese mice
Splenocytes from naïve mice were cultured with the specified acylcarnitines or vehicle and stimulated with anti-CD3, anti-CD28, and IL-2 for 48 hr. (a) CD69 expression on CD8 T cells cultured in 60 µM CAR8:0, CAR18:0, or vehicle. Viability of lymphocytes cultured with CAR8:0 (b), CAR18:0 (c), or vehicle. d, e, Splenocytes were cultured with the specified acylcarnitines or vehicle and stimulated with IL-2, IL-12, and IL-15. (d) IFN-γ expression by NK cells cultured in 60 µM CAR8:0, CAR18:0, or vehicle for 20 hr. (e) c-Myc expression by NK cells cultured in 60 µM CAR8:0, CAR18:0, or vehicle for 4 hr. f-h, Splenocytes were cultured with the specified acylcarnitines or vehicle and stimulated with anti-CD3, anti-CD28, and IL-2 for 48 hr. (f) MitoTracker Green (MTG) MFI of CD8 T cells cultured with 60 µM CAR8:0, CAR18:0, or vehicle, normalised to correct for intra-assay variability. Representative histogram (g) and summary data (h) of MitoSOX staining for mitochondrial reactive oxygen species in CD8 T cells cultured with 60 µM CAR8:0 (blue), CAR18:0 (orange), or vehicle (dark gray) and stimulated with anti-CD3, anti-CD28, and IL-2, or vehicle alone (light gray). i, GzmB expression by CD8 T cells treated with 30 µM CAR8:0, CAR18:0, or vehicle and stimulated with anti-CD3, CD28, and IL-2 for 48 hr. (a) n = 6; (b, c) n = 14-20, data combined from 4 independent experiments; (d) n = 5-6, data pooled from 3 independent experiments; (e) n = 8, data pooled from 2 independent experiments; (f) n = 3-4; (g,h) n = 5-6; data pooled from 4 independent experiments; (i) n = 7 human donors, 4 independent experiments. In bar graphs, bar height represents mean with error bars ± SEM. Comparisons were made using one-way ANOVA followed by Tukey’s multiple comparisons test (a-f, h) or repeated measures one-way ANOVA followed by Tukey’s multiple comparisons test (i).
