Development of Inhibitors of Supressors of Innate Immune Responses

哺乳动物细胞中Toll样受体(TLRs)的发现使 对先天免疫的理解。这些分子构成了模式识别膜 识别致病物质的分子，包括内毒素、鞭毛蛋白、蛋白多糖、脂磷壁酸 酸性、分支杆菌LAM、双链RNA和细菌CpG。衍生产品的范围 从革兰氏阳性和阴性细菌，分枝杆菌，真菌和病毒产品都激活 TLR通路。作为TLR激活的结果，可以激活适应性免疫的细胞因子 也会产生，因此它们是先天免疫和获得性免疫之间的桥梁。尽管 这些受体的关键作用，它们参与防御选择的药物较少明确。 在这项建议中，我们计划直接研究TLRs及其下游监管机构的角色 以防御一种选定的病原体，鼠疫耶尔森氏菌。我们将重点介绍已知的TLR及其 下游监管机构以及我们最近确定的一个新的下游监管机构， 君主-1蛋白。Monch-1下调TLR反应，包括NF-.B/AP-1激活，以及 肿瘤坏死因子-。回应。因此，它构成了一种新的TLR反应抑制因子，可能会影响 预防自然感染和接种疫苗的免疫。相应地，目标是(1)生产细胞 利用干扰RNA(RNAi)在TLR途径中缺乏已知和新的分子 ：技术学)。我们计划培养君主1号有缺陷的单核细胞U937细胞以及 选定的TLR及其下游信号包括TRAF2、IRAKs、MyD88和MD-2。(2)至 确定移除目标1中的靶向基因是否会导致对细胞的反应发生变化 鼠疫耶尔森氏菌的产物。(3)进一步了解Monch-1如何抑制炎症反应 通过测定Monch-1蛋白复合体的成分。

The discovery of Toll-like receptors (TLRs)in mammalians cells has revolutionized the understanding of innate immunity. These molecules constitute pattern recognition membrane molecules that recognize pathogenic products, including LPS, flagellin, proteoglycan, lipoteichoic acid, mycobacterium LAM, double stranded RNA, and bacterial CpG. The range of products derived from gram positive and negative bacteria, mycobacterium, fungi and viral products all activate the TLR pathways. As a consequence of TLR activation, cytokines that can activate adaptive immunity are also produced, thus they serve as a bridge between innate and adaptive immunity. Despite the pivotal role of these receptors, their involvement in defense against select agent is less well defined. In this proposal, we plan to directly examine the roles of TLRs and its downstream regulators in defense against a select agent, Yersinia pestis. We will focus on known TLRs and their downstream regulators as well as a new downstream regulator that we recently identified, the Monarch-1 protein. Monarch-1 downregulates TLR responses, including NF-.B/AP-1 activation, and TNF-. response. Thus its constitutes a novel suppressor of the TLR response that is likely to affect mmunity against r_atural infection and vaccination. Accordingly, the Aims are (1) To produce cell ines lacking known and new molecules in the TLR pathway by the use of interference RNA (RNAi :echnology). We plan to generate monocytic U937 cells that are defective in Monarch-1 as well as selected TLRs and their downstream signals including TRAF2, IRAKs, MyD88, and Md-2. (2) To determine if the removal of the genes targeted in Aim 1 causes alterations in responses to cellular products from Y. pestis. (3) To further understand how Monarch-1 inhibits inflammatory responses by determining components of the Monarch-1 protein complex.