FOXP3 promoter demethylation reveals the committed Treg population in humans.
FOXP3 promoter demethylation reveals the committed Treg population in humans.
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DOI：
10.1371/journal.pone.0001612
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发表时间：
2008-02-20
期刊：
影响因子：
3.7
通讯作者：
Winqvist O
中科院分区：
文献类型：
--
作者：
Janson PC;Winerdal ME;Marits P;Thörn M;Ohlsson R;Winqvist O
天然存在的胸腺源性调节性T细胞（TCFs）在维持自身耐受性中是中心的。转录因子FOXP 3对TcB的抑制活性至关重要，被认为是该人群最具特异性的标志物。然而，人类非调节性T细胞在激活后瞬时上调FOXP 3，这需要其他手段来鉴定Treg群体。由于表观遗传机制参与了细胞分化过程中稳定基因表达模式的建立，我们假设FOXP 3启动子的甲基化谱将允许区分真正定型的TcB。人CD 4 + CD 25 hi T细胞显示去甲基化的FOXP 3启动子（1.4%±0.95% SEM甲基化），而CD 4 + CD 25 lo T细胞部分甲基化（27.9%±7.1%）。此外，刺激的CD 4 + CD 25 lo T细胞瞬时表达FOXP 3，但仍保持部分甲基化，表明启动子甲基化是调节稳定FOXP 3表达和Treg定型的机制。此外，瞬时表达FOXP 3的细胞表现出与FOXP 3启动子的甲基化状态相关的抑制能力。作为亚硫酸氢盐测序的替代方案，我们提出了一种基于限制性酶的筛选方法来鉴定定型Treg，并应用该方法来评估各种培养条件的影响。我们发现，在活化后的长期培养中发生部分去甲基化，而加入TGF-β和/或IL-10不会诱导甲基化水平的任何额外变化。在Tumor中独特的FOXP 3启动子甲基化谱表明去甲基化模式是稳定FOXP 3表达和抑制表型的先决条件。目前，FOXP 3被用于鉴定几种人类疾病中的TcG，并且在免疫治疗中的过继性Treg转移具有未来的意义。在这些情况下，需要区分真正的T细胞与瞬时FOXP 3+激活的T细胞。我们提出的筛选方法允许这种区分，并能够鉴定适合于体外扩增和临床使用的细胞。
Naturally occurring thymus derived regulatory T cells (Tregs) are central in the maintenance of self-tolerance. The transcription factor FOXP3 is crucial for the suppressive activity of Tregs and is considered the most specific marker for this population. However, human non regulatory T cells upregulate FOXP3 transiently upon activation which calls for other means to identify the Treg population. Since epigenetic mechanisms are involved in the establishment of stable gene expression patterns during cell differentiation, we hypothesized that the methylation profile of the FOXP3 promoter would allow the distinction of truly committed Tregs. Human CD4+CD25hi Tregs displayed a demethylated FOXP3 promoter (1.4%±0.95% SEM methylated) in contrast to CD4+CD25lo T cells which were partially methylated (27.9%±7.1%). Furthermore, stimulated CD4+CD25lo T cells transiently expressed FOXP3 but remained partially methylated, suggesting promoter methylation as a mechanism for regulation of stable FOXP3 expression and Treg commitment. In addition, transient FOXP3 expressing cells exhibited suppressive abilities that correlate to the methylation status of the FOXP3 promoter. As an alternative to bisulphite sequencing, we present a restriction enzyme based screening method for the identification of committed Tregs and apply this method to evaluate the effect of various culturing conditions. We show that a partial demethylation occurs in long-term cultures after activation, whereas the addition of TGF-β and/or IL-10 does not induce any additional change in methylation level. The unique FOXP3 promoter methylation profile in Tregs suggests that a demethylated pattern is a prerequisite for stable FOXP3 expression and suppressive phenotype. Presently, FOXP3 is used to identify Tregs in several human diseases and there are future implications for adoptive Treg transfer in immunotherapy. In these settings there is a need to distinguish true Tregs from transiently FOXP3+ activated T cells. The screening method we present allows this distinction and enables the identification of cells suitable for in vitro expansions and clinical use.
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影响因子：
32.4
作者：
Fontenot, JD;Rasmussen, JP;Rudensky, AY
通讯作者：
Rudensky, AY
DOI：
10.1073/pnas.0509484103
发表时间：
2006-04-25
影响因子：
11.1
作者：
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通讯作者：
Rudensky, AY
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作者：
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影响因子：
5.4
作者：
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通讯作者：
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影响因子：
56.9
作者：
Hori, S;Nomura, T;Sakaguchi, S
通讯作者：
Sakaguchi, S