Figure 1.

Uptake and influence of exogenous D-2HG on survival, proliferation, and activation of T-cells. A) The uptake of D-2HG, exogenously supplied at different concentrations to T-cell cultures (stimulated with anti-CD2/CD3/CD28 coated beads), was measured after an incubation time of 72 h by a colorimetric enzymatic assay (Ai, n = 3). Additionally, intracellular total 2HG (D- and S-enantiomer) levels of T-cells isolated from healthy donors (HD) and AML patients (AML) were quantified by liquid chromatography-mass spectrometry (Aii). Cells were furthermore analyzed regarding the effects on proliferation (B; n = 6), survival (C; n = 11), T-cell receptor signaling (D; n = 4-7), and activation-related surface marker expression as measured by FACS (E; n = 10) upon D-2HG treatment. T-cells were either unstimulated (unstim, grey bars) or stimulated without (0 mM, black) or with (orange) D-2HG at indicated concentrations. FACS plots show analyses from a representative experiment. The Western Blot image shows two representative donors from a total of four. * p < 0.05; ** p < 0.01; ns: not significant; n.d.: not detected.