Extended Data Fig. 8: Guanidine-treatment of MDSCs abrogates their suppressive activity on CD8 T cell effector functions.
From: Regulatory myeloid cells paralyze T cells through cell–cell transfer of the metabolite methylglyoxal
a-c and e,f, human CD8 T cells were cultures in the presence of MDSCs that were partially pretreated with indicated compounds. a, T cells were stimulated alone, treated with DMBG, in the presence of MDSCs (pretreated with DMBG, methylguanidine, aminoguanidine or rodenidine (200 µM each). Proliferation (a) or cytotoxic activity (b) was measured by the dilution of CFSE (n = 3) or by ELISpot (n = 3 biological independent samples). d, Suppressive activity of CD11b+Ly6C+ or CD11b+Ly6G+ cells from the central nerves system during the recovery phase of experimental autoimmune encephalomyelitis (EAE; day 22 after immunization partially pretreated with DMBG (n = 3). e activated CD8 T cells were cocultured with CD4+CD25+CD127- regulatory T cells. Were Indicated, DMBG (200 µM) were added and proliferation was measured on day 3 (n = 3 biological independent samples). f, CD15+ cells were isolated from blood from the same patient were isolated an cocultured with CFSE labeled, activated CD8 T cells. Proliferation was measured by the dilution of CFSE (n = 2 biological independent samples). g, h, T cells were cocultivated with MDSCs for 30 min and reisolated. The glucose uptake capacity (2-NBDG) was measured at 30 min intervals (n = 3 biological independent samples). h, continuous glucose uptake measurement after addition of DMBG (n = 3), live-dead discrimination of CD8 T cells cultured alone, with monocytes or with MDSCs (n = 5 biological independent samples). j, cell count of CD8 T cells cultured alone, with monocytes or with MDSCs (n = 3 biological independent samples). *p < 0.05; **p < 0.01; ***p < 0.001; two-way unpaired t-test.
