DNA methylation changes at human Th2 cytokine genes coincide with DNase I hypersensitive site formation during CD4+ T cell differentiation
DNA methylation changes at human Th2 cytokine genes coincide with DNase I hypersensitive site formation during CD4+ T cell differentiation
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DOI：
10.4049/jimmunol.169.4.1893
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发表时间：
2002-08-15
影响因子：
4.4
通讯作者：
Staynov, DZ
中科院分区：
文献类型：
--
作者：
Santangelo, S;Cousins, DJ;Staynov, DZ
幼稚CD 4（+）T淋巴细胞分化为Th 1和Th 2谱系产生细胞或体液免疫应答。Th 2细胞表达细胞因子IL-4、IL-5和IL-13，这些细胞因子与哮喘和特应性有关。关于鼠Th 2细胞因子表达的调节已经发表了很多，但是在人原代T细胞中的研究不太常见。我们已经开发了一种方法，用于将人CD 45 RA（+）（幼稚）T细胞分化为Th 1和Th 2群体，这些群体显示出不同的细胞因子表达谱。我们使用亚硫酸氢盐DNA修饰和测序检测了CpG甲基化，以及在人T细胞分化前后和正常人皮肤成纤维细胞中IL-4和IL-13基因周围的染色质结构。在幼稚细胞中，DNA主要是甲基化的。Th 2分化后，IL-4和IL-13上出现DNase I超敏位点（DHS），而CpG去甲基化仅发生在Th 2特异性DHS周围。DHS和CpG去甲基化与Th 2特异性转录因子加塔-3的共有结合位点一致。虽然成纤维细胞，如幼稚和Th 1细胞，不表达IL-4或IL-13，但观察到不同于Th 2特异性位点的DHS和未甲基化CpG位点，表明该簇中的染色质结构不仅根据IL-4/IL-13表达在T细胞中变化，而且具有组织特异性。
The differentiation of naive CD4(+) T lymphocytes into Th1 and Th2 lineages generates either cellular or humoral immune responses. Th2 cells express the cytokines IL-4, -5, and -13, which are implicated in asthma and atopy. Much has been published about the regulation of murine Th2 cytokine expression, but studies in human primary T cells are less common. We have developed a method for differentiating human CD45RA(+) (naive) T cells into Th1 and Th2 populations that display distinct cytokine expression profiles. We examined both CpG methylation, using bisulfite DNA modification and sequencing, and chromatin structure around the IL-4 and IL-13 genes before and after human T cell differentiation and in normal human skin fibroblasts., In naive cells, the DNA was predominantly methylated. After Th2 differentiation, DNase I hypersensitive sites (DHS) appeared at IL-4 and IL-13 and CpG demethylation occurred only around the Th2-specific DHS. Both DHS and CpG demethylation coincided with consensus binding sites for the Th2-specific transcription factor GATA-3. Although fibroblasts, like naive and Th1 cells, did not express IL-4 or IL-13, DHS and unmethylated CpG sites that were distinct from the Th2-specific sites were observed, suggesting that chromatin structure in this cluster not only varies in T cells according to IL-4/IL-13 expression but is also tissue specific.