Induction of the Unfolded Protein Response by Brucella abortus VceC

描述(申请人提供)：先天免疫系统通过对保守的微生物结构的模式识别来检测组织中微生物的存在，称为病原体相关分子模式(PAMPs)。然而，PAMPs可以存在于所有微生物中，无论它们的致病潜力如何。为了将病原体与其他致病潜力较低的微生物区分开来，先天性免疫系统可以通过尚未完全解决的机制来检测病原体诱导的过程，例如宿主细胞胞浆中微生物产物的存在。由于病原菌表达的PAMPs可以并行激活许多模式识别受体，因此识别参与检测病原体诱导过程的信号通路通常是困难的。流产布鲁氏菌是一种隐形病原体，表达修饰的PAMPs，不再作为模式识别受体的激动剂。因此，在流产杆菌感染期间产生的宿主反应完全依赖于检测作为病原体诱导的过程的毒力因子--IV型分泌系统(T4SS)的部署。在这里，我们建议使用这个模式生物来定义一个新的信号通路，它涉及到将T4SS依赖的蛋白质注入宿主细胞胞浆作为病原体诱导的过程。我们的长期目标是确定在流产杆菌感染过程中诱导炎性宿主反应的机制和后果。这项应用的目的是研究T4SS底物VCEC移位到宿主细胞中如何诱导促炎反应并改变疾病结局。我们假设T4SS底物VCEC的易位激活了未折叠蛋白反应(UPR)，从而诱导了依赖于核因子B的炎症反应，从而促进了流产。我们将测试我们的假说的关键方面，并通过追求两个特定的目标来实现这一应用的目标：(1)阐明检测VCEC靶向内质网的信号通路；(2)表征UPR激活体内炎症反应的机制。这项工作的成功完成将推动这一领域的向前发展，建立UPR作为先天性免疫系统的一个组成部分，该系统检测作为病原体诱导过程的针对ER的微生物蛋白。这一新概念不仅对细菌发病机制有重要影响，而且对宿主-病毒相互作用、先天免疫和某些炎症性疾病的发病机制也有重要影响，如1型糖尿病和炎症性肠病。

DESCRIPTION (provided by applicant): The innate immune system detects the presence of microbes in tissue by pattern recognition of conserved microbial structures, known as pathogen-associated molecular patterns (PAMPs). However PAMPs can be present in all microbes regardless of their pathogenic potential. To distinguish pathogens from other microbes with lower disease-causing potential the innate immune system can detect pathogen-induced processes, such as the presence of microbial products in the host cell cytosol, through mechanisms that are not fully resolved. Identification of signaling pathways involved in the detection of pathogen-induced processes is often difficult because PAMPs expressed by a pathogen can activate many pattern recognition receptors in parallel. Brucella abortus is a stealthy pathogen expressing modified PAMPs that no longer serve as agonists for pattern recognition receptors. As a result, host responses generated during B. abortus infection are entirely dependent on detecting the deployment of a virulence factor, the type IV secretion system (T4SS), as a pathogen-induced process. Here we propose to use this model organism to define a new signaling pathway involved in sensing the T4SS-dependent injection of proteins into the host cell cytosol as a pathogen-induced process. Our long-range goal is to determine the mechanisms and consequences of inducing inflammatory host responses during B. abortus infection. The objectives of this application are to study how translocation of the T4SS substrate VceC into host cells induces pro-inflammatory responses and alters the disease outcome. We hypothesize that translocation of the T4SS substrate VceC activates the unfolded protein response (UPR) with consequent induction of NF-?B-dependent inflammatory responses, thereby contributing to B. abortus- induced abortion. We will test key aspects of our hypothesis and accomplish the objectives of this application by pursuing two specific aims: (1) Elucidate the signaling pathway that detects targeting of VceC to the endoplasmic reticulum; and (2) Characterize the mechanism by which the UPR activates inflammatory responses in vivo. Successful completion of this work will move the field forward by establishing the UPR as a component of the innate immune system that detects microbial proteins targeting the ER as a pathogen- induced process. This new concept has important ramifications not only for bacterial pathogenesis, but also for host-virus interactions, innate immunity and the pathogenesis of certain inflammatory disorders, such as type 1 diabetes and inflammatory bowel disease.