Generation of MAVS conditional KO mice to study cell-type specific immunity

摘要 西尼罗河病毒(West Nile Virus，WNV)是一种嗜神经性的黄病毒，是B类NIAID的优先制剂，是引起 美国由蚊子传播的人类脑炎。西尼罗河病毒的持续传播，与 由于缺乏对抗或预防感染的特定疗法或疫苗，迫切需要确定 控制感染和免疫的病毒和宿主过程。西尼罗河病毒在人类中的致病机制很差。 定义，但一个西尼罗河病毒感染的小鼠模型，忠实地概括了西尼罗河病毒的主要阶段 在人类中观察到的发病机制，为导致西尼罗河病毒的机制提供了重要的见解 疾病。先天免疫反应(由RIG-I样受体[RLR]信号介导)和体液和 细胞介导的反应对于预防西尼罗河病毒感染至关重要。RLR信号通路的功能 在西尼罗河病毒感染期间触发抗病毒免疫防御和程序保护性免疫。我们的研究已经 最近发现了RLR信号与CD8+T细胞免疫调节之间的新联系 病毒感染。为了支持我们的发现，其他研究小组现在已经证实了MAV，即中枢适配器蛋白 RLR介导的先天免疫信号所必需的，在慢性疾病过程中调节CD8+T细胞反应 病毒感染，在细菌感染过程中驱动CD4+T细胞极化，并调节TH1和TH17 自身免疫性脑炎期间的反应，从而表明RLR信号在脑内的重要性 编程T细胞免疫。然而，MAVS调节T细胞的免疫学机制 豁免权还没有得到很好的理解。我们试图通过开发一种新的研究工具来填补我们知识上的这一空白 以细胞和组织特异性的方式研究MAV的免疫调节。在目标1中，我们将产生Mavsfl/fl小鼠 这可以与表达cre的小鼠杂交，以上下文相关的方式抑制MAV的表达。在……里面 目的：建立树突状细胞表达MAVS缺失的MAVS-DC KO和MAVS-CD8KO小鼠 细胞(DC)和CD8+T细胞。我们将使用这些小鼠来评估对西尼罗河病毒的保护 感染、病毒致病机制的控制和CD8+T细胞免疫反应的发展。结果来自于 本研究将揭示MAVS介导的T细胞免疫调节的免疫学机制 西尼罗河病毒感染期间的免疫力。这些研究可能会发现治疗和免疫调节 RLR信号通路在病毒感染和疫苗接种过程中的潜力。 ！！

Abstract West Nile virus (WNV), a category B NIAID priority agent, is a neurotropic flavivirus that is the leading cause of mosquito-borne encephalitis of humans in the United States. The continuing spread of WNV, combined with the lack of specific therapeutics or vaccines to combat or prevent infection, imparts a pressing need to identify the viral and host processes that control infection and immunity. The pathogenesis of WNV in humans is poorly defined, but a mouse model of WNV infection, which faithfully recapitulates the major phases of WNV pathogenesis observed in humans, has provided significant insights into the mechanisms that cause WNV disease. The innate immune response (mediated by RIG-I like receptor [RLR] signaling) and the humoral and cell-mediated responses are critical for protection against WNV infection. The RLR signaling pathway functions to trigger antiviral immune defenses and program protective immunity during WNV infection. Our studies have recently revealed a novel connection between RLR signaling and regulation of CD8+ T cell immunity during virus infection. In support of our findings, other groups have now implicated MAVS, the central adaptor protein required for RLR-mediated innate immune signaling, with regulation of CD8+ T cell responses during chronic viral infection, driving CD4+ T cell polarization during bacterial infection, and regulating TH1 and TH17 responses during autoimmune encephalitis, thus demonstrating the importance of RLR signaling in programming T cell immunity. However, the immunological mechanism underlying MAVS regulation of T cell immunity are not well understood. We seek to fill this gap in our knowledge by developing a new research tool to study MAVS immune regulation in a cell and tissue-specific manner. In Aim 1, we will generate Mavsfl/fl mice that can be crossed with cre-expressing mice to ablate MAVS expression in a context-dependent manner. In Aim 2, we will generate MAVS-DC KO and MAVS-CD8 KO mice, which lack MAVS expression in dendritic cells (DCs) and CD8+ T cells, respectively. We will use these mice to evaluate protection against WNV infection, control of viral pathogenesis, and development of CD8+ T cell immune responses. The results from this study will reveal the immunological mechanisms underlying MAVS-mediated immune regulation of T cell immunity during WNV infection. These studies will potentially uncover the therapeutic and immune-modulating potential of the RLR signaling pathway during virus infection and vaccination. !!