Deficiency in KLF2 or S1P1 leads to impaired thymic egress (3, 8, 39). zinc finger transcription factor KLF2. Our findings have identified 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. whole-body KO mice die shortly after birth (28, 29). Although JMJD3 has been implicated in the thymic egress (30), its target genes and the regulatory mechanisms in T cell trafficking have not been reported. Here, we show that T cell Desacetyl asperulosidic acid trafficking from the thymus to the spleen and lymph nodes (LNs) was markedly altered in promoter. Specifically, our results indicate that JMJD3 regulates the expression of cytoskeletal PDLIM4 by stabilizing the KLF2-ASH2L complex and thus controls T cell trafficking. Results Critical role of JMJD3 in T cell trafficking from the thymus to spleen and LNs. We previously generated T cellCspecific deletion of (mice with CD4-Cre mice and found that JMJD3 plays a critical role in T cell differentiation (28). To further define the role of JMJD3 in the homeostasis of T cell populations and trafficking, we performed flow cytometric analysis of T cells isolated from the thymus, spleen, and LNs. While thymic CD4 SP T cells and CD8 SP T cells were dramatically increased, the percentages of CD4+ T cells were markedly decreased (approximately 50%) in both spleens and LNs of results in marked accumulation of thymic CD4 and CD8 SP T cells, reducing their ability to migrate from the thymus to secondary lymphoid organs. Open in a separate window Figure 1 Analysis of T cell populations in different organs of WT and = 8/group, from 2 independent experiments). Data are reported as mean + SD from 3 independent experiments. **< 0.01, Students test. (C) Percentages of CD4+ and CD8+ cells in Desacetyl asperulosidic acid the peripheral blood from WT and = 4/5. **< 0.01, Students test. NS, no significant differences. Trafficking of WT and JMJD3-deficient T cells in adoptive Desacetyl asperulosidic acid transfer models. Next, we sought to determine the intrinsic trafficking properties of WT and mice, with or without CD4-Cre to generate 2D2:and 2D2:(WT) and 2D2:deficiency causes defects in T cell migration.(A) Thymic CD4 SP cells from either WT or = 5) (upper panel). Absolute numbers of CD4+ cells in the spleens of recipient or 2D2:= 5). Absolute numbers of CD4+ TCRV3.2+ TCRV11+ cells in the spleens and LNs of recipient C57BL/6 mice were determined 24 hours after adoptive transfer (left segment) and positive cell ratio (right segment) by FACS. Data are plotted as mean + SD from 3 independent experiments. *< 0.05, **< 0.01, Students test. Next, we sought to determine whether the reduced trafficking ability of Jmjd3-deficient CD4+ T cells has functional consequences for the induction of experimental autoimmune encephalomyelitis (EAE), a T cellCmediated autoimmune disease of Rabbit Polyclonal to eNOS the CNS. WT and and CD4-Cre mice to generate TRP-1:and TRP-1:(WT) and TRP-1:were downregulated, while and were upregulated in thymic and were remarkably downregulated in and (encoding the PDZ and LIM domain protein 4), but not = 4. *< 0.05; **< 0.01, Students test. (C) CD4+ T cells from 2D2:(WT) mice or 2D2:or = 4). Absolute numbers of TCRV3.2+/V11+GFP+CD4+ T cells in spleens and LNs were determined by flow cytometry 48 hours after adoptive Desacetyl asperulosidic acid transfer. Desacetyl asperulosidic acid Data are presented as mean + SD from 3 independent experiments. **< 0.01, 1-way ANOVA with Tukeys multiple comparisons test. (D) WT and = 3/group; 1 experiment. (E) Thymic CD4 SP T cells were isolated from WT mice. 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,.