CD8+ T cell–derived IL-13 increases macrophage IL-10 to resolve neuropathic pain
Susmita K. Singh, Karen Krukowski, Geoffroy O. Laumet, Drew Weis, Jenolyn F. Alexander, Cobi J. Heijnen, Annemieke Kavelaars
Susmita K. Singh, Karen Krukowski, Geoffroy O. Laumet, Drew Weis, Jenolyn F. Alexander, Cobi J. Heijnen, Annemieke Kavelaars
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Research Article Neuroscience

CD8+ T cell–derived IL-13 increases macrophage IL-10 to resolve neuropathic pain

Abstract

Understanding the endogenous mechanisms regulating resolution of pain may identify novel targets for treatment of chronic pain. Resolution of chemotherapy-induced peripheral neuropathy (CIPN) after treatment completion depends on CD8+ T cells and on IL-10 produced by other cells. Using Rag2–/– mice lacking T and B cells and adoptive transfer of Il13–/– CD8+ T cells, we showed that CD8+ T cells producing IL-13 were required for resolution of CIPN. Intrathecal administration of anti–IL-13 delayed resolution of CIPN and reduced IL-10 production by dorsal root ganglion macrophages. Depleting local CD206+ macrophages also delayed resolution of CIPN. In vitro, TIM3+CD8+ T cells cultured with cisplatin, apoptotic cells, or phosphatidylserine liposomes produced IL-13, which induced IL-10 in macrophages. In vivo, resolution of CIPN was delayed by intrathecal administration of anti-TIM3. Resolution was also delayed in Rag2–/– mice reconstituted with Havcr2 (TIM3)–/– CD8+ T cells. Our data indicated that cell damage induced by cisplatin activated TIM3 on CD8+ T cells, leading to increased IL-13 production, which in turn induced macrophage IL-10 production and resolution of CIPN. Development of exogenous activators of the IL-13/IL-10 pain resolution pathway may provide a way to treat the underlying cause of chronic pain.

Authors

Susmita K. Singh, Karen Krukowski, Geoffroy O. Laumet, Drew Weis, Jenolyn F. Alexander, Cobi J. Heijnen, Annemieke Kavelaars

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Figure 5

TIM3 on CD8+ T cells is required for induction of IL-13 production by cisplatin in vitro.

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TIM3 on CD8+ T cells is required for induction of IL-13 production by ci...
(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.