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Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T-cell differentiation.

Li Q, Zou J, Wang M, Ding X, Chepelev I, Zhou X, Zhao W, Wei G, Cui J, Zhao K, Wang HY, Wang RF - Nat Commun (2014)

Bottom Line: The skewing of T-cell differentiation is concomitant with changes in the expression of key transcription factors and cytokines.H3K27me3 and H3K4me3 levels in Jmjd3-deficient cells are correlated with altered gene expression through interactions with specific transcription factors.Our results identify Jmjd3 as an epigenetic factor in T-cell differentiation via changes in histone methylation and target gene expression.

View Article: PubMed Central - PubMed

Affiliation: Center for Inflammation and Epigenetics, Houston Methodist Research Institute, Houston, Texas 77030, USA.

ABSTRACT
Epigenetic factors have been implicated in the regulation of CD4(+) T-cell differentiation. Jmjd3 plays a role in many biological processes, but its in vivo function in T-cell differentiation remains unknown. Here we report that Jmjd3 ablation promotes CD4(+) T-cell differentiation into Th2 and Th17 cells in the small intestine and colon, and inhibits T-cell differentiation into Th1 cells under different cytokine-polarizing conditions and in a Th1-dependent colitis model. Jmjd3 deficiency also restrains the plasticity of the conversion of Th2, Th17 or Treg cells to Th1 cells. The skewing of T-cell differentiation is concomitant with changes in the expression of key transcription factors and cytokines. H3K27me3 and H3K4me3 levels in Jmjd3-deficient cells are correlated with altered gene expression through interactions with specific transcription factors. Our results identify Jmjd3 as an epigenetic factor in T-cell differentiation via changes in histone methylation and target gene expression.

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Jmjd3 ablation alters the differentiation of naïve T cells into Th1, Th2, Th17, and Treg cell lineages in vitro(a and b) Frequency of IFN-γ, IL-4, and IL-17-producing and Foxp3-expressing CD4+ T cells after exposure of WT and Jmjd3 cKO naïve CD4+ T cells to ThN, Th1, Th2, Th17, or Treg-inducing. Mean percentage of the indicated T cell populations ± SD shown as histograms (bottom panel). (c) IFN-γ (top panel) and IL-4 (bottom panel) production by WT and Jmjd3 cKO naïve CD4+ T cells stimulated with soluble anti-CD3 and anti-CD28 in the presence of T cell-depleted irradiated splenocytes. (d) IL-4 production by WT and Jmjd3 cKO naïve CD4+ T cells under Th2-stimulating cell culture conditions with exogenous addition of IFN-γ. (e) Frequency of Th2 cells after exposure to Th2-stimulating cell culture conditions plus exogenous IFN-γ. (f) Relative gene expression of T cell differentiation factors from WT and Jmjd3 cKO naïve CD4+ cells cultured under ThN, Th1, Th2, Th17, or Treg-inducing conditions. X axis, different cytokine condition; Y axis, relative expression at mRNA level. (g) Intracellular staining of T-bet and Gata3 in CD4+ T cells cultured under ThN conditions (representative of three independent experiments, n = 3 for each group,*p < 0.05, **p < 0.01 determined by Student's t-test).
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Figure 2: Jmjd3 ablation alters the differentiation of naïve T cells into Th1, Th2, Th17, and Treg cell lineages in vitro(a and b) Frequency of IFN-γ, IL-4, and IL-17-producing and Foxp3-expressing CD4+ T cells after exposure of WT and Jmjd3 cKO naïve CD4+ T cells to ThN, Th1, Th2, Th17, or Treg-inducing. Mean percentage of the indicated T cell populations ± SD shown as histograms (bottom panel). (c) IFN-γ (top panel) and IL-4 (bottom panel) production by WT and Jmjd3 cKO naïve CD4+ T cells stimulated with soluble anti-CD3 and anti-CD28 in the presence of T cell-depleted irradiated splenocytes. (d) IL-4 production by WT and Jmjd3 cKO naïve CD4+ T cells under Th2-stimulating cell culture conditions with exogenous addition of IFN-γ. (e) Frequency of Th2 cells after exposure to Th2-stimulating cell culture conditions plus exogenous IFN-γ. (f) Relative gene expression of T cell differentiation factors from WT and Jmjd3 cKO naïve CD4+ cells cultured under ThN, Th1, Th2, Th17, or Treg-inducing conditions. X axis, different cytokine condition; Y axis, relative expression at mRNA level. (g) Intracellular staining of T-bet and Gata3 in CD4+ T cells cultured under ThN conditions (representative of three independent experiments, n = 3 for each group,*p < 0.05, **p < 0.01 determined by Student's t-test).

Mentions: To determine whether Jmjd3 ablation affects CD4+ T cell differentiation under different cytokine-polarizing conditions, the percentage of IFN-γ-, IL-4-, and IL-17-producing T cells and Foxp3-expressing T cells was analyzed in WT and Jmjd3 cKO purified naïve CD4+ T cells cultured under ThN (non-skewing cytokines), Th1 (in the presence of IL-12), Th2 (in the presence of IL-4), Th17 (in the presence of transforming growth factor [TGF-β] and IL-6), and Treg cell (in the presence of TGF-β and IL-2) conditions for 4 days. Jmjd3 ablation reduced the percentage of IFN-γ-producing Th1 cells from 50.1% to 6.2% and increased IL-4-producing Th2 cells from 3.2% to 48.6% under ThN conditions (Fig. 2a). Under Th1 conditions, Jmjd3 ablation also reduced the percentage of IFN-γ-producing Th1 cells. The percentage of T cells producing both IFN-γ and IL-4 was slightly higher in Jmjd3-deficient T cells compared with WT T cells (10.3% vs. 6.2%) under ThN conditions and significantly higher under Th1-stimulating cell culture conditions (11.3% vs. 2.2%) (Fig. 2a), suggesting that Jmjd3-deficient T cells are readily differentiated into Th2 cells. Under Th2 conditions, Jmjd3-deficient T cells slightly increased in their differentiation into IL-4 producing Th2 cells and slightly decreased into IFN-γ-producing Th1 cells or into IFN-γ- and IL-4-producing cells. However, these changes were not statistically significant, suggesting that exogenous cytokines under Th2 conditions may mask the intrinsic difference between WT and Jmjd3-deficient T cells. Th17 cell differentiation was increased in Jmjd3-deficient T cells under Th17-stimulating conditions (Fig. 2b). In contrast, the inducible Treg cell population was markedly reduced under Treg cell conditions in Jmjd3-deficient T cells compared with WT T cells (34.3% vs. 63.1%) (Fig. 2b). Jmjd3 ablation affects cytokine production of freshly isolated naïve CD4+ T cells stimulated with anti-CD3 and anti-CD28. IFN-γ production was significantly reduced in Jmjd3-deficient CD4+ T cells at 12, 24, and 48 h (p < 0.01), whereas IL-4 production was markedly increased at 24 and 48 h (p < 0.05 and p < 0.01, respectively) (Fig. 2c). To determine whether the increased IL-4 production was due to the early decreased IFN-γ production, we treated cultured T cells with exogenous IFN-γ and measured IL-4 production. Exogenous IFN-γ reduced IL-4 production in both WT and Jmjd3-deficient T cells; however, the Jmjd3-deficient T cells still produced higher amounts of IL-4 compared with WT T cells (Fig. 2d), suggesting that Jmjd3-deficient T cells preferentially polarize to Th2. Similarly, exogenous IFN-γ suppressed IL-4 producing Th2 cells in both WT and Jmjd3-deficient T cells. However, exogenous IFN-γ suppressed IL-4 producing Th2 cells to a greater extent in WT compared with Jmjd3-deficient T cells (Fig. 2e). Taken together, these data indicate that Jmjd3 deficiency results in a Th1 to Th2 shift in the CD4+ T cell population.


Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T-cell differentiation.

Li Q, Zou J, Wang M, Ding X, Chepelev I, Zhou X, Zhao W, Wei G, Cui J, Zhao K, Wang HY, Wang RF - Nat Commun (2014)

Jmjd3 ablation alters the differentiation of naïve T cells into Th1, Th2, Th17, and Treg cell lineages in vitro(a and b) Frequency of IFN-γ, IL-4, and IL-17-producing and Foxp3-expressing CD4+ T cells after exposure of WT and Jmjd3 cKO naïve CD4+ T cells to ThN, Th1, Th2, Th17, or Treg-inducing. Mean percentage of the indicated T cell populations ± SD shown as histograms (bottom panel). (c) IFN-γ (top panel) and IL-4 (bottom panel) production by WT and Jmjd3 cKO naïve CD4+ T cells stimulated with soluble anti-CD3 and anti-CD28 in the presence of T cell-depleted irradiated splenocytes. (d) IL-4 production by WT and Jmjd3 cKO naïve CD4+ T cells under Th2-stimulating cell culture conditions with exogenous addition of IFN-γ. (e) Frequency of Th2 cells after exposure to Th2-stimulating cell culture conditions plus exogenous IFN-γ. (f) Relative gene expression of T cell differentiation factors from WT and Jmjd3 cKO naïve CD4+ cells cultured under ThN, Th1, Th2, Th17, or Treg-inducing conditions. X axis, different cytokine condition; Y axis, relative expression at mRNA level. (g) Intracellular staining of T-bet and Gata3 in CD4+ T cells cultured under ThN conditions (representative of three independent experiments, n = 3 for each group,*p < 0.05, **p < 0.01 determined by Student's t-test).
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Figure 2: Jmjd3 ablation alters the differentiation of naïve T cells into Th1, Th2, Th17, and Treg cell lineages in vitro(a and b) Frequency of IFN-γ, IL-4, and IL-17-producing and Foxp3-expressing CD4+ T cells after exposure of WT and Jmjd3 cKO naïve CD4+ T cells to ThN, Th1, Th2, Th17, or Treg-inducing. Mean percentage of the indicated T cell populations ± SD shown as histograms (bottom panel). (c) IFN-γ (top panel) and IL-4 (bottom panel) production by WT and Jmjd3 cKO naïve CD4+ T cells stimulated with soluble anti-CD3 and anti-CD28 in the presence of T cell-depleted irradiated splenocytes. (d) IL-4 production by WT and Jmjd3 cKO naïve CD4+ T cells under Th2-stimulating cell culture conditions with exogenous addition of IFN-γ. (e) Frequency of Th2 cells after exposure to Th2-stimulating cell culture conditions plus exogenous IFN-γ. (f) Relative gene expression of T cell differentiation factors from WT and Jmjd3 cKO naïve CD4+ cells cultured under ThN, Th1, Th2, Th17, or Treg-inducing conditions. X axis, different cytokine condition; Y axis, relative expression at mRNA level. (g) Intracellular staining of T-bet and Gata3 in CD4+ T cells cultured under ThN conditions (representative of three independent experiments, n = 3 for each group,*p < 0.05, **p < 0.01 determined by Student's t-test).
Mentions: To determine whether Jmjd3 ablation affects CD4+ T cell differentiation under different cytokine-polarizing conditions, the percentage of IFN-γ-, IL-4-, and IL-17-producing T cells and Foxp3-expressing T cells was analyzed in WT and Jmjd3 cKO purified naïve CD4+ T cells cultured under ThN (non-skewing cytokines), Th1 (in the presence of IL-12), Th2 (in the presence of IL-4), Th17 (in the presence of transforming growth factor [TGF-β] and IL-6), and Treg cell (in the presence of TGF-β and IL-2) conditions for 4 days. Jmjd3 ablation reduced the percentage of IFN-γ-producing Th1 cells from 50.1% to 6.2% and increased IL-4-producing Th2 cells from 3.2% to 48.6% under ThN conditions (Fig. 2a). Under Th1 conditions, Jmjd3 ablation also reduced the percentage of IFN-γ-producing Th1 cells. The percentage of T cells producing both IFN-γ and IL-4 was slightly higher in Jmjd3-deficient T cells compared with WT T cells (10.3% vs. 6.2%) under ThN conditions and significantly higher under Th1-stimulating cell culture conditions (11.3% vs. 2.2%) (Fig. 2a), suggesting that Jmjd3-deficient T cells are readily differentiated into Th2 cells. Under Th2 conditions, Jmjd3-deficient T cells slightly increased in their differentiation into IL-4 producing Th2 cells and slightly decreased into IFN-γ-producing Th1 cells or into IFN-γ- and IL-4-producing cells. However, these changes were not statistically significant, suggesting that exogenous cytokines under Th2 conditions may mask the intrinsic difference between WT and Jmjd3-deficient T cells. Th17 cell differentiation was increased in Jmjd3-deficient T cells under Th17-stimulating conditions (Fig. 2b). In contrast, the inducible Treg cell population was markedly reduced under Treg cell conditions in Jmjd3-deficient T cells compared with WT T cells (34.3% vs. 63.1%) (Fig. 2b). Jmjd3 ablation affects cytokine production of freshly isolated naïve CD4+ T cells stimulated with anti-CD3 and anti-CD28. IFN-γ production was significantly reduced in Jmjd3-deficient CD4+ T cells at 12, 24, and 48 h (p < 0.01), whereas IL-4 production was markedly increased at 24 and 48 h (p < 0.05 and p < 0.01, respectively) (Fig. 2c). To determine whether the increased IL-4 production was due to the early decreased IFN-γ production, we treated cultured T cells with exogenous IFN-γ and measured IL-4 production. Exogenous IFN-γ reduced IL-4 production in both WT and Jmjd3-deficient T cells; however, the Jmjd3-deficient T cells still produced higher amounts of IL-4 compared with WT T cells (Fig. 2d), suggesting that Jmjd3-deficient T cells preferentially polarize to Th2. Similarly, exogenous IFN-γ suppressed IL-4 producing Th2 cells in both WT and Jmjd3-deficient T cells. However, exogenous IFN-γ suppressed IL-4 producing Th2 cells to a greater extent in WT compared with Jmjd3-deficient T cells (Fig. 2e). Taken together, these data indicate that Jmjd3 deficiency results in a Th1 to Th2 shift in the CD4+ T cell population.

Bottom Line: The skewing of T-cell differentiation is concomitant with changes in the expression of key transcription factors and cytokines.H3K27me3 and H3K4me3 levels in Jmjd3-deficient cells are correlated with altered gene expression through interactions with specific transcription factors.Our results identify Jmjd3 as an epigenetic factor in T-cell differentiation via changes in histone methylation and target gene expression.

View Article: PubMed Central - PubMed

Affiliation: Center for Inflammation and Epigenetics, Houston Methodist Research Institute, Houston, Texas 77030, USA.

ABSTRACT
Epigenetic factors have been implicated in the regulation of CD4(+) T-cell differentiation. Jmjd3 plays a role in many biological processes, but its in vivo function in T-cell differentiation remains unknown. Here we report that Jmjd3 ablation promotes CD4(+) T-cell differentiation into Th2 and Th17 cells in the small intestine and colon, and inhibits T-cell differentiation into Th1 cells under different cytokine-polarizing conditions and in a Th1-dependent colitis model. Jmjd3 deficiency also restrains the plasticity of the conversion of Th2, Th17 or Treg cells to Th1 cells. The skewing of T-cell differentiation is concomitant with changes in the expression of key transcription factors and cytokines. H3K27me3 and H3K4me3 levels in Jmjd3-deficient cells are correlated with altered gene expression through interactions with specific transcription factors. Our results identify Jmjd3 as an epigenetic factor in T-cell differentiation via changes in histone methylation and target gene expression.

Show MeSH
Related in: MedlinePlus