JMJD3 regulates CD4+ T cell trafficking by targeting actin cytoskeleton regulatory gene Pdlim4
Chuntang Fu, … , Helen Y. Wang, Rong-Fu Wang
Chuntang Fu, … , Helen Y. Wang, Rong-Fu Wang
Published November 1, 2019; First published August 8, 2019
Citation Information: J Clin Invest. 2019;129(11):4745-4757. https://doi.org/10.1172/JCI128293.
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Categories: Research Article Autoimmunity Cell biology Immunology

JMJD3 regulates CD4+ T cell trafficking by targeting actin cytoskeleton regulatory gene Pdlim4

Abstract

Histone H3K27 demethylase JMJD3 plays a critical role in gene expression and T cell differentiation. However, the role and mechanisms of JMJD3 in T cell trafficking remain poorly understood. Here, we show that JMJD3 deficiency in CD4+ T cells resulted in an accumulation of T cells in the thymus and reduction of T cell number in the secondary lymphoid organs. We identified PDLIM4 as a significantly downregulated target gene in JMJD3-deficient CD4+ T cells by gene profiling and ChIP-Seq analyses. We further showed that PDLIM4 functioned as an adaptor protein to interact with sphingosine-1 phosphate receptor 1 (S1P1) and filamentous actin (F-actin), thus serving as a key regulator of T cell trafficking. Mechanistically, JMJD3 bound to the promoter and gene-body regions of the Pdlim4 gene and regulated its expression by interacting with zinc finger transcription factor KLF2. Our findings have identified Pdlim4 as a JMJD3 target gene that affects T cell trafficking by cooperating with S1P1 and have provided insights into the molecular mechanisms by which JMJD3 regulates genes involved in T cell trafficking.

Authors

Chuntang Fu, Qingtian Li, Jia Zou, Changsheng Xing, Mei Luo, Bingnan Yin, Junjun Chu, Jiaming Yu, Xin Liu, Helen Y. Wang, Rong-Fu Wang

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Figure 3

Identification of the JMJD3 target genes in CD4+ T cells and functional rescue of T cell defects by ectopic expression of Pdlim4.

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Identification of the JMJD3 target genes in CD4+ T cells and functional ...
(A) Heatmap from microarray analysis of upregulated and downregulated genes in WT and Jmjd3-cKO thymic CD4 SP T cells. (B) Real-time PCR analysis of a panel of genes between WT and Jmjd3-cKO thymic CD4 SP T cells. Expression levels are given as the ratio of the target gene to the control gene to correct for variations in the starting amount of mRNA (gene/Gapdh ×1000). n = 4. *P < 0.05; **P < 0.01, Student’s t test. (C) CD4+ T cells from 2D2:Jmjd3fl/fl (WT) mice or 2D2:Jmjd3-cKO mice were activated with MOG35–55 peptide in vitro before transduction with GFP-expressing retroviral vectors containing Jmjd3 or Pdlim4. Equal numbers of GFP+CD4+ T cells were intravenously injected into sublethally irradiated C57BL/6 mice (n = 4). Absolute numbers of TCRVα3.2+/Vβ11+GFP+CD4+ T cells in spleens and LNs were determined by flow cytometry 48 hours after adoptive transfer. Data are presented as mean + SD from 3 independent experiments. **P < 0.01, 1-way ANOVA with Tukey’s multiple comparisons test. (D) WT and Jmjd3-cKO bone marrow cells overexpressing control GFP or Pdlim4-GFP were transplanted into lethally irradiated C57BL/6 (WT) mice to generate chimeric mice. Flow cytometric analysis of CD4+ and CD8+ T cells from the thymus and the spleens of chimeric mice. n = 3/group; 1 experiment. (E) Thymic CD4 SP T cells were isolated from WT mice. Pdlim4 KO was generated using the CRISPR-Cas9 system. Cells were labeled with CFSE and then intravenously injected into sublethally irradiated C57BL/6 mice. After 48 hours, spleens, LNs, and peripheral blood were analyzed by flow cytometry for CD4+ and CFSE-stained cells. Experiments were repeated 3 independent times. n = 3/group; 1 experiment.