Induction of 6-sulfated glycans with cell adhesion activity via T-bet and GATA-3 in human helper T cells
Induction of 6-sulfated glycans with cell adhesion activity via T-bet and GATA-3 in human helper T cells
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通过 T-bet 和 GATA-3 在人类辅助 T 细胞中诱导具有细胞粘附活性的 6-硫酸化聚糖
DOI：
10.1016/j.bbagen.2012.03.005
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Kannagi R.
中科院分区：
文献类型：
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作者：
Sakuma K;Chen GY;Aoki M;Kannagi R.
细胞表面6-硫酸化聚糖通过细胞间相互作用在各种免疫学事件中发挥重要作用。6-硫酸化过程由6-磺基转移酶家族同工酶介导。我们先前证明GlcNAc 6ST-1是一种同工酶基因，在人辅助性T（Th）细胞中由加塔-3和NF-κB诱导。方法采用5 ′-RACE分析、染色质免疫沉淀和报告基因分析等方法研究HEC-GlcNAc 6ST的转录调控机制。进行RNA敲低和强制表达实验以证明HEC-GlcNAc 6ST对6-硫酸化聚糖表达的贡献。我们鉴定了HEC-GlcNAc 6ST启动子中Sp1、T-bet和加塔-3的潜在结合位点。报告分析表明，转染Sp1增强的活性，而光神霉素A，Sp1特异性抑制剂，抑制它。转染T-bet增加的活性，这是通过引入一个突变到潜在的T-bet结合位点抑制。单独的加塔-3不能提高活性，尽管共转染蛋白激酶A（已知其通过加塔-3的磷酸化增强Th 2细胞中的IL-5转录）引起活性升高。RNA敲低和Jurkat细胞中HEC-GlcNAc 6ST的强制表达分别下调和上调α 2，6-唾液酸化6-磺基N-乙酰乳糖胺，这是B细胞特异性CD 22抗原的优先配体。从这些结果中，我们得出结论，T-bet和加塔-3以及Sp1控制聚糖的表达与细胞粘附活性，通过调节HEC-GlcNAc 6ST转录在Th cells.GENERAL SIGNIFICANCE这些结果可能提供了一个线索，生物调节Th细胞与选择素和其他碳水化合物识别分子的T-bet和加塔-3。
BACKGROUNDCell surface 6-sulfated glycans play important roles in various immunological events through cell-to-cell interactions. The 6-sulfation process is mediated by 6-sulfotransferase family isoenzymes. We previously demonstrated that GlcNAc6ST-1, one of the isoenzyme genes, is induced by GATA-3 and NF-κB in human helper T (Th) cells. However, transcriptional regulation of HEC-GlcNAc6ST, another isoenzyme important in Th cells, remains unclear.METHODS5′-RACE analysis, chromatin immunoprecipitation, and reporter assays were performed to reveal transcriptional regulation of HEC-GlcNAc6ST. RNA-knockdown and forced expression experiments were performed to demonstrate the contribution of HEC-GlcNAc6ST to the 6-sulfated glycan expression.RESULTSWe identified potential binding sites of Sp1, T-bet, and GATA-3 in the HEC-GlcNAc6ST promoter. Reporter assays indicated that transfection of Sp1 enhanced the activity, whereas mithramycin A, an Sp1-specific inhibitor, repressed it. Transfection of T-bet increased the activity, which was inhibited by introducing a mutation into the potential T-bet binding site. GATA-3 alone could not elevate the activity, although co-transfection of protein kinase A, which is known to enhance IL-5 transcription in Th2 cells through phosphorylation of GATA-3, caused elevation. RNA-knockdown and forced expression of HEC-GlcNAc6ST in Jurkat cells down- and up-regulated α2,6-sialylated 6-sulfo N-acetyllactosamine, a preferential ligand for B-cell-specific CD22 antigen, respectively. From these results, we concluded that T-bet and GATA-3 as well as Sp1 control the expression of glycan with cell-adhesion activity by regulating HEC-GlcNAc6ST transcription in Th cells.GENERAL SIGNIFICANCEThese results may provide a clue to biological regulation of Th-cell interaction with selectins and other carbohydrate-recognition molecules by T-bet and GATA-3.