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

摘要 西尼罗河病毒（WNV），一种B类NIAID优先因子，是一种嗜神经黄病毒，其是引起感染性疾病的主要原因。 蚊子传播的人类脑炎。西尼罗河病毒的持续传播， 由于缺乏对抗或预防感染的特异性治疗剂或疫苗，迫切需要确定 病毒和宿主控制感染和免疫的过程。西尼罗河病毒在人类中的发病机制很差， 定义，但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，为引起西尼罗河病毒的机制提供了重要的见解 疾病先天性免疫应答（由RIG-I样受体[RLR]信号传导介导）和体液免疫应答， 细胞介导的应答对于防止WNV感染是至关重要的。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 最近揭示了RLR信号传导和CD 8 + 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涉及MAVS，中央衔接蛋白 TLR介导的先天性免疫信号传导所需，在慢性炎症期间调节CD 8 + T细胞应答 病毒感染，在细菌感染期间驱动CD 4 + T细胞极化，并调节TH 1和TH 17 在自身免疫性脑炎期间的反应，从而证明了RLR信号转导在免疫性脑炎中的重要性。 编程T细胞免疫。然而，MAVS调节T细胞的免疫学机制可能与MAVS调节T细胞的免疫学机制有关。 对免疫性了解不多。我们试图通过开发一种新的研究工具来填补我们知识的空白 以细胞和组织特异性方式研究MAVS免疫调节。在目标1中，我们将产生Mavsfl/fl小鼠 其可以与表达cre的小鼠杂交，以上下文依赖的方式消除MAVS表达。在 目的2，我们将产生MAVS-DC KO和MAVS-CD 8 KO小鼠，其在树突状细胞中缺乏MAVS表达。 细胞（DC）和CD 8 + 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作用 感染、控制病毒发病机制以及CD 8 + T细胞免疫反应的发展。的结果 本研究将揭示MAVS介导的T细胞免疫调节的免疫学机制。 感染西尼罗河病毒期间的免疫力。这些研究将有可能揭示治疗和免疫调节 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. !!