TLR2/NLR Signal Regulation of Protective Immunity to F. Tularensis

项目总结（见说明）; Toll样受体（TLR）和NOD样受体（NLR）在先天性感染反应中的信号传导 影响宿主细胞对细菌胞质复制的容许性， 保护性免疫然而，目前还不完全清楚这些信号事件是如何通过参与细胞内的 TLR和NLR通过以下途径影响赋予针对致死性细菌感染的保护的能力： 预防针还不清楚抗体如何以失活的抗体形式结合土拉热弗朗西丝菌（Ft）。 本提案中评价的疫苗原（1，2），即FF-单克隆抗体（IFf-mAb）复合物， 通过TLR 2或NLR的先天性信号传导，从而产生适应性/保护性免疫应答。 病原体抵抗氧化还原介导的宿主防御或改变基于氧化还原的宿主细胞的能力的变化 信号传导，如已经显示的Ft的超氧化物歧化酶缺陷的活疫苗株（LVS） 活性（3，4）影响TLR和NLR依赖性炎症反应。因此，病原体来源的 抗氧化剂可能影响不同疫苗原产生适应性/保护性免疫的能力。 因此，我们假设通过TLR 2和/或NLR转导的信号影响免疫调节剂的功效。 疫苗接种后，这些信号受到Ft和/或细菌的抗体调理作用的影响。 促炎能力。该假设将通过以下方式进行检验：1）确定mAb结合Ft 和Ft抗氧化剂突变体在先天免疫（诱导）阶段对协同TLR 2/NLR信号传导的影响 分子水平的抗Ft免疫应答和2）评估TLR 2和/或NLR信号传导是否 影响用各种Ff制剂接种后保护性免疫的诱导。 协同作用：子项目2中获得的结果将为TLR和NLR导向的先天性 在次级项目1中采用的现行疫苗战略的背景下， 这些方法。参与巨噬细胞ROS/RNS产生的信号通路与这些通路交叉， 通过FcR、TLR和NLR刺激激活，即PI 3激酶、NF-κ B和MAPK途径; 因此，这些目标的研究结果，对于以下方面的实验具有重要的指导意义，并将有助于为以下方面的实验提供信息 次级项目3.此外，这种协同作用将通过提供表征的抗氧化剂突变体来促进 从子项目3，从而使我们的子项目阐明Ft抗氧化途径如何影响

PROJECT SUMMARY (See instructions); Toll-like receptor (TLR) and NOD-like receptor (NLR) signaling during the innate response to infection influences the host cell's permissiveness for cytosolic replication of bacteria and the development of protective immunity. However, it is not entirely clear how these signaling events initiated by engagement of TLRs and NLRs influences the capacity to confer protection against lethal bacterial infection through vaccination. It also is unclear how antibody bound to Francisella tularensis (Ft) in the form of an inactivated Ff-monoclonal antibody (IFf-mAb) complex, the vaccinogen (1,2) under evaluation in this proposal, alters innate signaling via TLR2 or NLRs and thereby the adaptive/protective immune response. Changes in a pathogen's capacity to resist redox-mediated host defenses or alter redox-based host cell signaling, as has been shown for the live vaccine strain (LVS) of Ft deficient for superoxide dismutase activity (3, 4), impacts both TLR and NLR-dependent inflammatory responses. Thus, pathogen-derived antioxidants likely influence the ability of different vaccinogens to engender adaptive/protective immunity. Therefore, we hypothesize that signals transduced via TLR2 and/or NLRs influence the efficacy of vaccination and these signals are impacted by antibody opsonization of Ft and/or the bacterium's proinflammatory capacity. This hypothesis will be tested by 1) Establishing the impact of mAb bound Ft and Ft antioxidant mutants on synergistic TLR2/NLR signaling during the innate immune (inductive) phase of the anti-Ft immune response at the molecular level and 2) Evaluating whether TLR2 and/or NLR signaling impacts induction of protective immunity after vaccination with various Ff formulations. Synergy: Results obtained in Subproject 2 will provide a new view of TLR and NLR-directed innate responses in the context of the ongoing vaccine strategies employed in Subproject 1, enabling refinement of these approaches. The signaling pathways involved in macrophage ROS/RNS production intersect those activated by FcR, TLR, and NLR stimulation, namely the PI3 kinase, NF-KB, and MAPK pathways; accordingly, the results of these Aims are significantly relevant to, and will help inform, experiments in Subproject 3. Further, this synergy will be facilitated by the provision of characterized antioxidant mutants from Subproject 3, thus enabling our Subproject to shed light on how Ft antioxidant pathways impact