Regulation of IL-3 and GM-CSF genes and their receptors

1.细胞因子基因在T细胞中的调控。利用TH1， TH2和TH0表型的人和小鼠T细胞克隆，我们一直在研究IL-3， IL-4， IL-5和GM-CSF基因的坐标和差异表达。小鼠GM-CSF启动子的CLE2/GC-box和CLE0在响应PMA和Ca离子载体的转录激活中是必不可少的。CLE2定义NF-kappaB的结合位点。主要GC-box结合活性A1被纯化，鉴定为Sp1。我们从活化的Jurkat提取物中纯化了一个人NF-AT蛋白，并发现它与GM-CSF启动子内的CLE0紧密结合，并与AP1相关。我们分离出一种具有锌指基序的新蛋白，它与IL-3启动子的富含CT/ gc的区域结合。我们还在IL-5、IL-4和IL-2启动子中发现了顺式调控元件。这些工作是与博士合作完成的。Naoko Arai和de Vries Jan在dnax。受体和信号转导。我们一直在研究GM-CSF/IL-3受体的结构和功能。GM-CSF受体的下游有几个信号通路。betac的膜近端区域对增殖、c-myc和pim-1诱导以及Jak2关联至关重要。betac的c端区在抑制细胞凋亡、Ras、Raf、MAPK活化和c-fos、c-jun诱导等方面具有重要作用。我们还发现，从表达hGM-CSF受体的转基因小鼠中提取的骨髓细胞在hGM-CSF作用下具有增殖能力，可诱导所有髓系细胞。此外，我们还产生了携带β - c和β -3零突变的小鼠。Betac突变小鼠也表现出肺病理，包括淋巴细胞浸润和类似肺泡蛋白沉积的区域。这些工作是与DNAX的宫岛atsushi博士和汉堡大学的Wolfram博士合作完成的。

1.Regulation of cytokine genes in T cells. Using human and mouse T cell clones with TH1, TH2, and TH0 phenotype, we have been working on the coordinate and differential expression of the IL-3, IL-4, IL-5 and GM-CSF genes. The CLE2/GC-box and CLE0 of the mouse GM-CSF promoter are essential for transcriptional activation in response to PMA and Ca ionophore. CLE2 defines a binding site for NF-kappaB.The major GC-box binding activity A1 was purified and was identified as Sp1. We purified a human NF-AT protein from activated Jurkat extract and showed that it strongly bound to the CLE0 within the GM-CSF promoter in association with AP1. We isolated a novel protein with zinc finger motifs which binds to the CT/GC-rich region of the IL-3 promoter. We also identified cis-regulatory elements within IL-5, IL-4 and IL-2 promoters. These works were done in collaboration with Drs.Naoko Arai and de Vries Jan in DNAX.2.Receptor and Signal transduction. We have been working on the structure and function of GM-CSF/IL-3 receptors. There are several signal pathways downstream of the GM-CSF receptor. The membrane proximal region of betac is essential for proliferation, c-myc and pim-1 induction and Jak2 association. The C-terminal region of betac is important for the suppression of apoptosis, Ras, Raf, MAPK activation and c-fos, c-jun induction. We have also shown that bone marrow cells derived from transgenic mice which constitutively express hGM-CSF receptor have the proliferative capacity to induce all the myeloid cell lineages in response to hGM-CSF.Furthermore, we have generated mice carrying a null mutation of betac and betaIL-3. betac mutant mice also showed lung pathology consisting of lymphocytic infiltration and areas resembling alveolar proteinosis. These works were done in collaboration with Dr.Atsushi Miyajima in DNAX and Dr.Ostertag Wolfram in Hamburg University.