Redox Control of F. tularensis Pathogenesis

说明)： 我们的总体目标是开发有效接种肺图拉热症疫苗的方法。 在上一个供资期间，我们取得了相当大的进展，包括证明 对C57BL/6小鼠对高毒力A型图拉氏杆菌SchuS4株的显著保护作用 可以由I.N诱导。接种FCR靶向灭活LV(Ft)。这是我们接到的第一起案件 对灭活疫苗的了解提供了对Ft SchuS4的保护。在这 更新应用后，我们将进一步优化黏膜疫苗接种条件，表征疫苗的效果 肺疫苗接种，并确定负责保护性免疫的细胞和体液机制 对抗高毒力的A型毒株SchuS4。具体来说，我们将：1)确定I.N.的能力。 在没有或存在外源IL-12的情况下，接种FCR靶向免疫原，以增强 对粘膜的免疫反应和对粘膜的保护水平(I.N.)挑战Ft SchuS4。二 将使用不同的方法将靶向Fcr：a)给药预制mAb-IFF复合体， 和b)同时接种未复合Ft和抗Ff单抗；2)建立机制 负责通过评估FCR靶向组织的分布来增强免疫诱导 和淋巴器官，以确定‘\ft在次级淋巴组织和APC中的增强定位 在这些组织中，由于FCR靶向而发生。还将确定是否增强\fT APC上靶向FCR的处理和呈现结果；3)建立效应器机制 负责在I.N后加强保护。接种以FCR为靶标的细菌。其作用机制 粘膜接种后负责保护的人将通过被动转移抗F/进行调查 抗体或细胞给幼鼠，特别注意检查潜在的协同作用要求 体液免疫和细胞免疫机制之间的关系，以诱导有效的保护。的角色 诱导期和效应期的TLR/NLR和ROS/RNS将通过遗传方法进行检测 缺陷小鼠和特定的激动剂/拮抗剂，与分项目2和3的pls协商。 子项目1的结果将允许设计有效的生物防御的新的粘膜疫苗接种策略 并将为肺免疫机制提供新的见解，包括 负责预防呼吸性图拉热症。 相关性(参见说明)： 这一分项目的结果将使设计有效的新的粘膜疫苗接种策略成为可能。 对强毒Ft感染的生物防御，将为肺免疫提供新的见解 负责预防呼吸性图拉热症的机制。

instmctions): Our overall objective is to develop approaches for effective vaccination against pulmonary tularemia. During the previous funding period, we made considerable progress, including the demonstration that significant protection in C57BL/6 mice against the highly virulent type A strain of F. tularensis {Ft), SchuS4, can be induced by i.n. inoculation of FcR-targeted inactivated LVS {\Ft). This is the first case to our knowledge in wliich an inactivated vaccine has provided protection against Ft SchuS4. In this renewal application, we will further optimize conditions for mucosal vaccination, characterize the effects of pulmonary vaccination, and define the cellular and humoral mechanisms responsible for protective immunity against the highly virulent type A strain, SchuS4. Specifically, we will: 1) Determine the ability of i.n. vaccination with FcR-targeted immunogens, in the absence or presence of exogenous IL-12, to enhance the immune response to, and levels of protection against, mucosal (i.n.) challenge with Ft SchuS4. Two separate approaches will be used to target \Ft to FcR: a) administration of preformed mAb-IFf complexes, and b) simultaneous inoculation of uncomplexed \Ft plus anti-Ff mAb; 2) Establish the mechanisms responsible for enhanced induction of immunity by assessing the distribution of FcR targeted \Ft to tissues and lymphoid organs to determine if enhanced localization of '\Ft to secondary lymphoid tissues, and APC within these tissues, occurs as a result of FcR targeting. It will also be determined if enhanced \Ft processing and presentation results from targeting to FcR on APC; and 3) Establish the effector mechanisms responsible for enhanced protection after i.n. vaccination with FcR-targeted bacteria. The mechanisms responsible for protection after mucosal vaccination will be investigated by passive transfer of anti-F/ antibody or cells to naive mice with particular attention paid to examining a potential requirement for synergy between humoral and cellular immune mechanisms for induction of effective protection. The roles of TLR/NLRs and ROS/RNS in both inductive and effector phases will be examined by using genetically deficient mice and specific agonists/antagonists, in consultation with the PLs of subprojects 2 and 3. The results of subproject 1 will allow the design of new mucosal vaccination strategies for effective biodefense against infection with virulent Ft and will provide novel insight into the pulmonary immune mechanisms that are responsible for protection against respiratory tularemia. RELEVANCE (See Instruct'ons): The results of this subproject will allow the design of new mucosal vaccination strategies for effective biodefense against infection with virulent Ft and will provide novel insight into the pulmonary immune mechanisms that are responsible for protection against respiratory tularemia.