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 6

DUSP8 production and DUSP8–Pur-α interaction are induced in Th9 cells of people with asthma and atopic dermatitis.

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DUSP8 production and DUSP8–Pur-α interaction are induced in Th9 cells of...
(A) Flow cytometry analyses of DUSP8-postivie T cells in the PBLs of 1 representative person from the healthy-control group (HC) and person with asthma. Data shown are mean ± SEM, n = 6 (right panel). There were no antibodies in Pacific Blue channel (left panel). FMO, fluorescence minus 1 (isotype control). (B) Flow cytometry analyses of DUSP8-postive and IL-9–producing T cells in the PBLs of a representative person from the healthy-control (HC) group and person with asthma. Data shown are mean ± SEM, n = 4. Isotype denotes the control group with isotype controls for anti-IL-9 and anti-DUSP8 antibodies. (C) Confocal microscopy of DUSP8, Pur-α, DAPI, and IL-9 in the peripheral blood T cells of 1 representative person from the HC group and person with asthma. Mean ± SEM of DUSP8 levels (green intensity) and Pur-α nuclear localization (red over blue, %) from 3 images are shown. Additional 11 people from the HC group, 23 people with asthma, and 7 people with atopic dermatitis (AD) are shown in Supplemental Figure 12. Scale bars: 25 μm. (D) Confocal microscopy of PLA for the DUSP8–Pur-α interaction in peripheral blood T cells, which were freshly isolated from 27 people with asthma and 18 people from the healthy-control group. One representative person from the HC group and person with asthma from Cohort 1 are shown (upper). One representative person from the HC group and person with AD from Cohort 2 are shown (lower). For PLA, red dots represent direct interaction signals. Scale bars: 50 μm. (E) Quantification plots of PLA signals (panel D) in asthma T cells from Cohort 1 (left), Cohort 2 (center), and the combination of 2 cohorts (right). (F) Quantification plots of PLA signals in AD T cells (panel D) from combination of 2 cohorts. *P < 0.05; **P < 0.01; ****P < 0.0001 (2-tailed Student’s t test).