Yamashita Masakatsu, Ukai-Tadenuma Maki, Miyamoto Takeshi, Sugaya Kaoru, Hosokawa Hiroyuki, Hasegawa Akihiro, Kimura Motoko, Taniguchi Masaru, DeGregori James, Nakayama Toshinori
PRESTO Project, Japan Science and Technology Corporation, Department of Immunology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana Chuo-ku, Chiba 260-8670, Japan.
J Biol Chem. 2004 Jun 25;279(26):26983-90. doi: 10.1074/jbc.M403688200. Epub 2004 Apr 15.
GATA3 expression is essential for type-2 helper T (Th2) cell differentiation. GATA3-mediated chromatin remodeling at the Th2 cytokine gene loci, including Th2-specific long range histone hyperacetylation of the interleukin (IL)-13/IL-4 gene loci, occurs in developing Th2 cells. However, little is known about the role of GATA3, if any, in the maintenance of established remodeled chromatin at the Th2 cytokine gene loci. Here, we established a Cre/LoxP-based site-specific recombination system in cultured CD4 T cells using a unique adenovirus-mediated gene transfer technique. This system allowed us to investigate the effect of loss of GATA3 expression in in vitro differentiated Th2 cells. After ablation of GATA3, we detected reduced production of all Th2 cytokines, increased DNA methylation at the IL-4 gene locus, and decreased histone hyperacetylation at the IL-5 gene locus but not significantly so at the IL-13/IL-4 gene loci. Thus, GATA3 plays important roles in the maintenance of the Th2 phenotype and continuous chromatin remodeling of the specific Th2 cytokine gene locus through cell division.
GATA3表达对于2型辅助性T（Th2）细胞分化至关重要。在发育中的Th2细胞中，会发生GATA3介导的Th2细胞因子基因位点的染色质重塑，包括白细胞介素（IL）-13/IL-4基因位点的Th2特异性长程组蛋白高乙酰化。然而，关于GATA3在维持Th2细胞因子基因位点已建立的重塑染色质中的作用（如果有），人们知之甚少。在此，我们使用独特的腺病毒介导的基因转移技术，在培养的CD4 T细胞中建立了基于Cre/LoxP的位点特异性重组系统。该系统使我们能够研究体外分化的Th2细胞中GATA3表达缺失的影响。在GATA3缺失后，我们检测到所有Th2细胞因子的产生减少，IL-4基因位点的DNA甲基化增加，IL-5基因位点的组蛋白高乙酰化减少，但在IL-13/IL-4基因位点没有显著变化。因此，GATA3在维持Th2表型以及通过细胞分裂对特定Th2细胞因子基因位点进行持续的染色质重塑中发挥重要作用。
