DUSP8 induces TGF-β–stimulated IL-9 transcription and Th9-mediated allergic inflammation by promoting nuclear export of Pur-α
Huai-Chia Chuang, … , Ming-Han Chen, Tse-Hua Tan
Huai-Chia Chuang, … , Ming-Han Chen, Tse-Hua Tan
Published November 1, 2023
Citation Information: J Clin Invest. 2023;133(21):e166269. https://doi.org/10.1172/JCI166269.
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Research Article Inflammation

DUSP8 induces TGF-β–stimulated IL-9 transcription and Th9-mediated allergic inflammation by promoting nuclear export of Pur-α

Abstract

Dual-specificity phosphatase 8 (DUSP8) is a MAPK phosphatase that dephosphorylates and inactivates the kinase JNK. DUSP8 is highly expressed in T cells; however, the in vivo role of DUSP8 in T cells remains unclear. Using T cell–specific Dusp8 conditional KO (T-Dusp8 cKO) mice, mass spectrometry analysis, ChIP-Seq, and immune analysis, we found that DUSP8 interacted with Pur-α, stimulated interleukin-9 (IL-9) gene expression, and promoted Th9 differentiation. Mechanistically, DUSP8 dephosphorylated the transcriptional repressor Pur-α upon TGF-β signaling, leading to the nuclear export of Pur-α and subsequent IL-9 transcriptional activation. Furthermore, Il-9 mRNA levels were induced in Pur-α–deficient T cells. In addition, T-Dusp8–cKO mice displayed reduction of IL-9 and Th9-mediated immune responses in the allergic asthma model. Reduction of Il-9 mRNA levels in T cells and allergic responses of T-Dusp8–cKO mice was reversed by Pur-α knockout. Remarkably, DUSP8 protein levels and the DUSP8–Pur-α interaction were indeed increased in the cytoplasm of T cells from people with asthma and patients with atopic dermatitis. Collectively, DUSP8 induces TGF-β–stimulated IL-9 transcription and Th9-induced allergic responses by inhibiting the nuclear translocation of the transcriptional repressor Pur-α. DUSP8 may be a T-cell biomarker and therapeutic target for asthma and atopic dermatitis.

Authors

Huai-Chia Chuang, Chia-Hsin Hsueh, Pu-Ming Hsu, Ching-Yi Tsai, Ying-Chun Shih, Hsien-Yi Chiu, Yi-Ming Chen, Wen-Kuang Yu, Ming-Han Chen, Tse-Hua Tan

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Figure 5

T cell–specific Dusp8 cKO mice are resistant to OVA-induced allergic asthma.

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T cell–specific Dusp8 cKO mice are resistant to OVA-induced allergic ast...
(A–F) Induction of OVA-induced asthma models in WT, T-Dusp8–cKO, or T-Dusp8 cKO/Pur-α heterozygous–KO (cKO/Pur-α+/–) mice. H&E-stained sections of the lung tissues from mice on day 26 of the OVA-induced asthma model (A). Mean ± SEM of infiltrating immune cells from 3 images are shown below. Scale bars: 250 μm. The cytokine levels in the BALFs of mice on day 26 of the OVA-induced asthma model were determined using ELISA assays (B). n = 4. The OVA-specific serum IgE levels of mice on day 26 were determined by ELISA assays (C). Data are presented relative to the basal levels from a WT mouse. n = 5. Confocal microscopy analyses of CD3 (green), IL-9 (red), and DAPI in the lung tissues from mice on day 26 of the OVA-induced asthma model (D). Scale bars: 50 μm. Confocal microscopy analyses of CD3 (green), Pur-α (red), and DAPI in the lung tissues from OVA-immunized mice (E). Scale bars: 10 μm. OVA-specific T cell–mediated cytokine production (F). T cells were isolated from the lymph nodes of OVA-immunized mice on day 21 after immunization, followed by in vitro stimulation with OVA for 72 hours. The mRNA levels of Il-9 and Il-4 were determined using real-time PCR and were normalized to Srp72 levels. n = 4 per group. WT (Dusp8fl/fl); T-Dusp8 cKO, T cell–specific Dusp8 cKO (Dusp8fl/fl;Cd4-Cre); cKO/Pur-α+/–, T-Dusp8 cKO/Pur-α heterozygous KO mice; OVA, ovalbumin. Mean ± SEM are shown. *P value < 0.05; **P value < 0.01; ***P value < 0.001 (1-way ANOVA and Tukey’s posthoc test). P value in blue, WT versus cKO; P value in green, cKO/Pur-α+/– versus cKO; NS, nonsignificant. Data shown are representative of 3 independent experiments.