Fig. 2: ACAT inhibition induces metabolic re-wiring of CD8+ T cells.
From: Targeting human Acyl-CoA:cholesterol acyltransferase as a dual viral and T cell metabolic checkpoint
a Neutral lipid droplets (LipidTOX staining) in unstimulated PBMC ±ACAT inhibition (K-604 or equivalent concentration of DMSO for 1 h). Representative histogram and summary data (n = 12). b Confocal microscopy visualising the immunological synapse between a CMV-specific CD8+ T cell (white dashed outline, identified by CTB and TCR staining) and CMV peptide pulsed T2 cell (APC, blue) with staining of GM1-enriched microdomains (identified by CTB staining, green) and αβTCR (red). White scale bar: 5 mm. c CTB staining on PBMC from patients with CHB after stimulation with HBc OLP ±ACAT inhibition (Avasimibe or DMSO for 8d). Representative flow cytometry plot and summary data (n = 10). d–f Representative phosphoflow cytometry plots/histograms and summary data to detect phosphorylated TCR signalling molecules pERK (n = 13; d), pAkt (n = 13; e), pS6 (n = 8; f) ±ACAT inhibition (Avasimibe or DMSO for 8d) and stimulation with aCD3/aCD28. g Real-time oxygen consumption rate (OCR) of PMA/Ionomycin-stimulated purified CD8+ T cells ±ACAT inhibition (Avasimibe or DMSO for 16 h, n = 6). Compounds were added as indicated (oligomycin (Oligo), FCCP, rotenone+antimycin A (Rot/AA)) to determine basal respiration, ATP production and maximal respiration. OCR time course: median and interquartile range. h Basal extracellular acidification rate (ECAR) and basal OCR/ECAR ratio of PMA/Ionomycin-stimulated purified CD8+ T cells ±ACAT inhibition (Avasimibe or DMSO for 16 h, n = 6). P values determined by Wilcoxon matched-pairs signed-rank test.
