Kaneko Takaaki, Hosokawa Hiroyuki, Yamashita Masakatsu, Wang Chrong-Reen, Hasegawa Akihiro, Kimura Motoko Y, Kitajiama Masayuki, Kimura Fumio, Miyazaki Masaru, Nakayama Toshinori
Department of Immunology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chiba 260-8670, Japan.
Mol Immunol. 2007 Mar;44(9):2249-56. doi: 10.1016/j.molimm.2006.11.004. Epub 2006 Dec 12.
The differentiation of mouse naïve CD4 T cells into type 2 helper (Th2) cells is accompanied by chromatin remodeling at the nucleosomes associated with the IL-4, IL-13 and IL-5 genes. However, little is known about how chromatin remodeling of these Th2 cytokine gene loci occurs in human Th2 cells. We herein established an in vitro culture system in which both Th1 and Th2 cells are efficiently differentiated from human peripheral blood naïve CD4 T cells. This system allowed us to investigate the chromatin status at the Th2 cytokine gene loci and the IFNgamma locus in human Th2 and Th1 cells, respectively. In typical individuals, the chromatin remodeling indicated by the induction of hyper-acetylation of histone H3 lysine 9 and hyper-methylation of histone H3 lysine 4 was induced at the whole Th2 cytokine gene loci in developing Th2 cells. We more precisely assessed the methylation status of histone H3 lysine 4 at the Th2 cytokine gene loci (IL-5 exon 3, IL-5 promoter, IL-5/RAD50 intergenic region, RAD50 promoter, CGRE, CNS1, IL-13 promoter, IL-4 promoter, and VA enhancer regions) and the IFNgamma locus in developing Th1 and Th2 cells prepared from 20 healthy volunteers. Th2-cell specific chromatin remodeling was induced at most of the Th2 cytokine gene loci. In parallel with the induction of chromatin remodeling, GATA3 mRNA was preferentially expressed in developing Th2 cells, whereas T-bet, HLX and ROG mRNA was selectively expressed in developing Th1 cells.
小鼠初始CD4 T细胞向2型辅助性（Th2）细胞的分化伴随着与白细胞介素4（IL-4）、白细胞介素13（IL-13）和白细胞介素5（IL-5）基因相关的核小体上的染色质重塑。然而，对于这些Th2细胞因子基因座在人类Th2细胞中如何发生染色质重塑知之甚少。我们在此建立了一种体外培养系统，其中Th1和Th2细胞都能从人外周血初始CD4 T细胞高效分化而来。该系统使我们能够分别研究人类Th2和Th1细胞中Th2细胞因子基因座以及干扰素γ（IFNγ）基因座的染色质状态。在典型个体中，发育中的Th2细胞在整个Th2细胞因子基因座上诱导出由组蛋白H3赖氨酸9的高乙酰化和组蛋白H3赖氨酸4的高甲基化所指示的染色质重塑。我们更精确地评估了从20名健康志愿者制备的发育中的Th1和Th2细胞中Th2细胞因子基因座（IL-5外显子3、IL-5启动子、IL-5/RAD50基因间区域、RAD50启动子、CGRE、CNS1、IL-13启动子、IL-4启动子和VA增强子区域）以及IFNγ基因座上组蛋白H3赖氨酸4的甲基化状态。在大多数Th2细胞因子基因座上诱导出了Th2细胞特异性染色质重塑。与染色质重塑的诱导同时，GATA3 mRNA在发育中的Th2细胞中优先表达，而T-bet、HLX和ROG mRNA在发育中的Th1细胞中选择性表达。
