ACCESSORY CELL ACTIVATION IN THE IMMUNE RESPONSE

这项建议的目的是阐明 激活或分化中基因诱导的最早阶段 单核细胞。主要的关注点将是基因调控，支付 特别注意可诱导转录因子负责 调节分化和细胞激活。方法是使用 白介素原1-β(IL1B)和少量其他诱导物 以单核细胞特异性基因为例研究其发病机制 单核细胞的即刻早期基因诱导。暂态评估 这些基因的转基因部分将与结合一起进行研究。 特定的蛋白质因子，以便深入了解其调控 机械装置。几种转录因子的cDNA序列 似乎对单核细胞的激活和分化很重要 用于旨在过度表达的共转染研究中，以便 评估具体效果。同样，这些基因的显性-负性突变 这些因素也将被调查。具体目标是：(一)澄清 白介素1B上游诱导的作用机制 以非细胞类型诱导IL1B为靶点的序列(UIS) (2)检测组织限制性SPI-I/PU-I的功能 转录因子，它与启动子近端序列结合。 IL1B基因和其他单核细胞特异性基因赋予组织特异性 表情。我们之前的研究表明，人类巨细胞病毒 (HCMV)IE1蛋白反式激活IL1B基因需要一个完整的 SPI-I/PU.I结合位点。因此，SPI-I/PU-I之间的关系 和HCMV IE1将利用这两个蛋白在功能上进行研究 和结构研究。

The object of this proposal is to elucidate the mechanisms for the earliest stages of gene induction in activated or differentiating monocytes. The primary focus will be on gene regulation, paying particular attention to inducible transcription factors responsible for mediating differentiation and cell activation. The approach is to use prointerleukin 1-beta (IL1B) and a small number of other inducible monocyte-specific genes as examples for investigating the mechanisms of immediate early gene induction in monocytes. Evaluation of transiently transfected portions of these genes will be studied along with the binding of specific protein factors in order to gain insight into the regulatory mechanisms. The cDNA sequences for several transcription factors which appear to be important for monocyte activation and differentiation will be used in cotransfection studies aimed at overexpression in order to evaluate specific effects. Similarly, dominant-negative mutants of these factors will also be investigated. Specific Aims are: (I) to elucidate the functional mechanism of action for the IL1B Upstream Induction Sequence (UIS) which targets cell type-independent induction of the IL1B gene; (2) to examine the function of the tissue-restricted Spi-I/PU.I transcription factor, which binds to the promoter proximal sequence in the IL1B gene and other monocyte-specific genes to confer tissue-specific expression. Our previous studies demonstrated that human cytomegalovirus (HCMV) IE1 protein trans-activation of the IL1B gene requires an intact Spi-I/PU.I binding site. Therefore, the relationship between Spi-I/PU.I and HCMV IE1 will be investigated using these two proteins in functional and structural studies.