NOVEL ZINC FINGER PROTEIN THAT INHIBITS TNF CYTOTOXICITY

细胞因子如肿瘤坏死因子（TNF）对肿瘤细胞具有多效性作用。 各种各样的细胞和组织。这些变化尤其 在TNF诱导粘附的内皮细胞中， 用于白细胞和增加膜促凝血活性。变化 这种性质可能是促炎作用的核心 的tnf为了阐明肿瘤坏死因子改变细胞凋亡的分子机制， 内皮细胞功能，我们利用差异斑块杂交， 鉴定TNF立即早期反应基因。七个这样的基因是 鉴定DNA测序显示，两个编码的已知细胞因子诱导 内皮细胞白细胞粘附分子-1和中性粒细胞趋化因子 因子一个编码核原癌基因c-jun，另一个编码 最近描述的单核细胞特异性趋化因子 与内皮细胞相关。单核细胞趋化因子的产生， 细胞因子激活的内皮细胞对于理解 血管壁在诸如动脉粥样硬化的疾病状态中的作用， 也可以部分解释TNF的间接血管生成活性。 值得注意的是，根据数据判断，其他三个cDNA是完全新的 部分DNA序列的数据库搜索重要的是，其中一个（A20） 含有七个锌指，这是某种类型的特征， 转录因子为了更好地理解单核细胞的作用， 化学引诱物和新的cDNA在血管壁疾病状态中的作用， 血管生成和炎症，我们提出以下研究： 目的1：（i）克隆编码新转录物的全长cDNA， 完成DNA测序具体目标2：（i）使细菌融合 单核细胞化学引诱物的蛋白质和新的 兔免疫和抗体产生。具体目标3：使用 在特异性目标2中产生的单特异性多克隆抗血清， 单核细胞化学引诱物和新抗原：a）体外，使用 细胞因子处理的内皮细胞和其它细胞。这将使亚细胞 新基因产物的定位。B）体内，炎症状态 例如伤口愈合和牛皮癣。对于单核细胞趋化因子， 对于检查动脉粥样硬化组织尤为重要。此外，本发明还 将进行原位杂交研究，以确定哪些细胞 在研究的组织中实际上是转录感兴趣的基因。

Cytokines such as tumor necrosis factor (TNF) exert pleiotropic effects on a wide variety of cells and tissues. These alterations are particularly profound in endothelial cells where TNF has been shown to induce adherence for leukocytes and to increase membrane procoagulant activity. Changes of this nature are likely to be central to the pro-inflammatory effects of TNF. In order to elucidate molecular mechanisms by which TNF alters endothelial cell function, we utilized differential plaque hybridization to identify TNF immediate early response genes. SEVEN such genes were identified. DNA sequencing revealed that two encoded known cytokine induced genes; endothelial leukocyte adhesion molecule-1 and neutrophil chemotactic factor. One encoded the nuclear protooncogene; c-jun, while another encoded a recently described monocyte specific chemotactic factor NOT previously associated with endothelium. The production of a monocyte chemotaxin by cytokine activated endothelium has important implications for understanding the role of the vessel wall in disease states such as atherosclerosis and may also in part explain the indirect angiogenic activity of TNF. Significantly, the other three cDNAs are completely novel as judged by data bank searches of partial DNA sequences. Importantly, one of these (A20) contains seven zinc fingers which is characteristic of a certain class of transcriptional factors. To better understand the role of the monocyte chemoattractant and the novel cDNAs in vessel wall disease states, angiogenesis and inflammation we propose the following studies: Specific Aim 1: (i) Clone full length cDNAs encoding the novel transcripts and complete their DNA sequencing. Specific Aim 2: (i) Make bacterial fusion protein for both the monocyte chemoattractant and the novel cDNAs for rabbit immunization and antibody production. Specific Aim 3: Using the monospecific polyclonal antiserum raised in Specific Aim 2, localize both the monocyte chemoattractant and the novel antigens: a) In Vitro, using cytokine treated endothelial and other cells. This will allow subcellular localization of the novel gene products. b) In vivo, in inflammatory states such as wound healing and psoriasis. For the monocyte chemoattractant it will be particularly important to examine atherosclerotic tissue. Further, in situ hybridization studies will be carried out to identify which cells in the tissue under study are actually transcribing the gene of interest.