Fig 1: TIM3 on CD8+ T cells is required for induction of IL-13 production by cisplatin in vitro.(A) TIM3+CD8+ T cells in the DRG during resolution of cisplatin-induced CIPN. The percentage of TIM3+CD8+ T cells in the DRG was quantified by flow cytometry. Welch’s t test; **P < 0.001; n = 4 male, 5 female/group. (B) BMDMs were cocultured with either WT CD8+ T cells or Havcr2 (TIM3)–/– CD8+ T cells. Cocultures were treated with cisplatin (1 µg/mL) or left untreated (unstim.) and 48 hours later, CD8+ T cells were assayed for IL-13 by flow cytometry. Two-way ANOVA, Holm-Šidák multiple-comparison test, **P < 0.05; n = 4 independent experiments; 2 male, 2 female. (C) Schematic representation of coculture experiment assessing effect of apoptotic cells on IL-13 production by T cells. WT mouse splenocytes were treated with cisplatin (2.5 µg/mL) for 24 hours or were UV exposed for 10 minutes and cultured for 24 hours. Apoptotic cells were cocultured with WT CD8+ T cells (D) or Havcr2 (TIM3)–/– CD8+ T cells (E). Cells were assayed for IL-13+CD8+ T cells by flow cytometry 48 hours later. One-way ANOVA, Tukey’s multiple-comparison test, *P < 0.05, **P <0.01, ***P < 0.001, ****P < 0.0001; n = 3–4 independent experiments; 2 male, 1–2 female. (F) WT mouse splenocytes were treated with PS (phosphatidylserine) or PC (phosphatidylcholine as negative control) liposomes at increasing concentrations. n = 4 independent experiments, 2 male, 2 female, assayed for IL-13+CD8+ T cells. (G) Cultures of WT mouse splenocytes with PS liposomes (50 µM) were treated with anti-TIM3 neutralizing antibody or IgG as control. n = 3–4 independent experiments; 2 male, 1 female. One-way ANOVA, Dunnett’s multiple-comparison test, *P < 0.05. (H) Havcr2 (TIM3)–/– splenocytes were treated with PS or PC (as negative control) liposomes at increasing concentrations. n = 3 independent experiments; 3 female. After 48 hours, the percentage of IL-13+CD8+ T cells was assessed by flow cytometry. All data are shown as mean ± SEM.