T cell–intrinsic STING inhibits cell cycle progression of T cells.

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T cell–intrinsic STING inhibits cell cycle progression of T cells. T cell–intrinsic STING inhibits cell cycle progression of T cells. (A) Expression of STING in BMDCs and CD4+ T cells by Western blot. (B) Naive CD8+ T cells from Sting+/+ or Sting−/− mice were stimulated with anti-CD3/CD28 Abs with or without graded amounts of cGAMP for 48 h, and cell growth was assessed by a WST-8 cell proliferation assay. (C) Naive CD4+ T cells from Sting+/− or Sting−/− OT-II Tg mice were cocultured with irradiated T cell–depleted splenocytes from C57BL/6 mice as APCs, together with OVA323–339 peptide (10 μM) in the presence or absence of graded amounts of cGAMP, and cell growth was assessed by a WST-8 cell proliferation assay. (D) Naive CD4+ T cells were pretreated with (open column) or without (closed column) Z-VAD and stimulated with anti-CD3/CD28 Abs with or without cGAMP or DMXAA for 48 h, and cell growth was assessed by a WST-8 cell proliferation assay. (E) Naive CD4+ T cells from Rip3+/+ (closed column) or Rip3−/− (open column) mice were stimulated with anti-CD3/CD28 Abs with or without cGAMP or DMXAA for 48 h, and cell growth was assessed by a WST-8 cell proliferation assay. (F) qPCR analysis of the expression of cell cycle–related genes in CD4+ T cells from Sting+/− (closed column) or Sting−/− (open column) mice upon stimulation with anti-CD3/CD28 Abs in the presence or absence of cGAMP (30 μg/ml) for 24 h. (G) RNA-seq data for the expression of cell cycle–related genes in CD4+ T cells stimulated with anti-CD3/CD28 Abs with or without cGAMP for 24 h. Data are the mean from duplicate (B–E) or triplicate (F) ± SD. Data are representative of at least three independent experiments. (A—F) *P < 0.05, t test (compared with WT cells treated with cGAMP). (E) *P < 0.05, t test (compared with control cells treated with etoposide). Takayuki Imanishi et al. LSA 2019;2:e201800282 © 2019 Imanishi et al.