Fig. 8.

Disruption glutaminolysis impacts on NV T-cell IL-2 production. IL-2 production of NV T-cells cultured with anti-CD3 (2 μg/mL) and anti-CD28 (20 μg/mL) in the presence or absence of a STAT5 inhibitor (STAT5i; 100 μM), b glutamine withdrawal (- Q). c Schematic of the different mechanisms used to investigate glutamine metabolism in human NV T-cells. IL-2 production of NV T-cells cultured with d DON (50 μM) or e aminooxyacetic acid (AOA; 0.25–1 mM). f NV T-cells activated and treated as in b with the addition of dimethyl 2-oxoglutarate (DMK; 0.3 mM) with downstream analysis of IL-2 production. NV T-cell cultured with g oligomycin (100 nM) and h BMS303141 (1 μM) with IL-2 production measured. i Schematic of NV T-cell activation via the T-cell receptor leading to downstream STAT5 phosphorylation. Glutaminolysis is regulated by STAT5 in an mTORC1-dependent manner. Statistical analysis was performed using an unpaired t-test (a, b, d, f, g), a Kruskal−Wallis test (e) or a one-way ANOVA (g). Data are representative of four experiments (a, d–e, h), five experiments (b) and three experiments (f) expressed as mean ± SEM; *p ≤ 0.05, **p ≤ 0.01