Figure 4. T_H17 cells preferentially maintain higher level of GSH than iT_reg cells.

(A–B) Naive CD4⁺ T cells from C57BL/6 mice were differentiated under iTreg or T_H17–inducing conditions and cells were collected at indicated times, followed by measuring intracellular GSH (A) and ROS (B) by FACS. (C) Naive CD4⁺ T cells from C57BL/6 mice were differentiated under T_H17 or iT_reg–inducing conditions for 5 days. The intracellular levels of GSH and GSSG were determined by mass spectrometry. (D) Naive CD4⁺T cells from WT and Gclm KO (top) or WT (CD4-Cre-, Gclc^fl/fl) and Gclc KO (CD4-Cre+, Gclc^fl/fl, (bottom) were differentiated under T_H17-inducing conditions for 5 days, followed by the measurement of GSH levels. Data in Figure 4A–D are representative of three independent experiments. Data represent the mean ± S.D.

Figure 4—figure supplement 1. De novo synthesis but not recycling of GSSG is required for providing GSH and suppressing ROS during TH17 cell differentiation.

(A) RNAs were isolated from cells differentiated under TH17 or iTreg-inducing conditions for indicated times, and used for real-time qPCR analyses of indicated genes. Expression levels in day 0 were set to 1 (B–D) Naive CD4+ T cells from WT and Gclm KO, or WT (CD4-Cre-, Gclc^fl/fl) and Gclc KO (CD4-Cre+, Gclc^fl/fl), or WT and Gsr KO mice were differentiated under TH17-inducing conditions for 5 days, followed by measuring ROS by FACS. Data in Figure B-D are representative of two independent experiments. Data represent the mean ±S.D.