Regulating Pathogen-induced Protective and Pathogenic CD8 T cells in the CNS

调节中枢神经系统中病原体诱导的保护性和致病性 CD8 T 细胞

• 批准号: 10722304
• 负责人: John T Harty
• 金额: $ 18.77万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-12 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10722304
• 关键词: Address; Area; Assessment tool; Attention; Brain; Brain Stem; CD8-Positive T-Lymphocytes; Cause of Death; Cells; Cellular biology; Central Nervous System Diseases; Central Nervous System Infections; Cerebral Malaria; Clinical; Data; Dependovirus; Disease; Dorsal; Etiology; Exploratory/Developmental Grant; Fostering; Generations; Genetic; Homeostasis; Human; Immune; Immune System Diseases; Immune system; Immunity; Immunization; Immunologics; Immunology; Infection; Injections; Knowledge; Ligands; LoxP-flanked allele; Lymphocytic choriomeningitis virus; Maintenance; Mediating; Meningeal lymphatic system; Modeling; Mus; Nature; Nervous System; Neurons; Operative Surgical Procedures; Organ; Outcome; Pain; Pathogenesis; Pathogenicity; Pathway interactions; Peripheral; Pharmaceutical Preparations; Process; Pruritus; Risk; Role; Serotonin; Signal Pathway; Signal Transduction; Specific qualifier value; Stimulus; Stress; System; T cell response; T-Lymphocyte; Testing; Tissues; Transgenic Organisms; Viral Encephalitis; Viral Pathogenesis; Virus; brain tissue; cell type; designer receptors exclusively activated by designer drugs; disability; draining lymph node; drinking water; experimental study; genetic technology; high reward; nervous system disorder; neuroimmunology; neurotransmission; novel therapeutics; pathogen; response; tissue resident memory T cell

Neurological diseases are a leading cause of death and disability, and despite diverse etiologies, share a common theme of immunological dysfunction. The immune and nervous systems are finely tuned to both sense and orchestrate responses to external stimuli. There are billions of neurons in the brain, yet we know next to nothing about how brain neurons interface with the immune system in the CNS. Recent advances in neuroimmunology include the discovery of meningeal lymphatic, roles for immune cells in CNS homeostasis and characterization of tissue resident memory T cells (Trm) that persist in the brain after CNS infection. We recently (Urban et al, Nature Immunology, 2020) provided evidence that Trm were also generated uniquely in the brain after peripheral immunizations and infections, suggesting that processes unique to the brain may foster Trm persistence in this organ after peripheral immunization. However, it is unknown if specific neuronal pathways in the brain contribute to Trm generation or persistence in this organ and whether neuronal signaling influences protection by CNS Trm. Several CNS diseases in humans have immunological etiologies, with T cells thought contribute to the pathogenesis of viral encephalitis (VE), cerebral malaria (CM) Thus, a second unknown is the role of brain neuronal signaling pathways in T cell-mediated diseases of the CNS. We will attack these problems using chemogenetics. Specifically, there are highly conserved neurons within the brainstem, such as the dorsal raphe (DR), which project broadly throughout the CNS and activate in response to immunity-relevant signals such as stress, pain, and itch. Transgenic B6 mice expressing SERT- cre target DR neurons. Stereotaxic injection of adeno-associated viruses expressing flox/stop designer receptors exclusively activated by designer drugs (DREADD), results in expression of the DREADD only in cre- expressing neurons. Transduced cre-expressing neurons are detected by mCherry expression and can be manipulated, depending on their nature (activating or inhibitory signals specified by the precise DREADD), by injection of a designer ligand. We have obtained these mice and virus systems and provided proof of our ability to perform the sophisticated stereotaxic surgeries and deliver the DREADD constructs to the targeted neurons. We will use the chemogenetic technology and our capacity to evaluate CNS immunity and immune-mediated pathogenesis to test the central hypothesis that brain neuronal signaling pathways contribute to the generation, maintenance and function of CNS Trm and the outcome of T cell-mediated CNS diseases. Specific Aim 1. Determine if DR neuronal signaling regulates the generation, maintenance or protective function of brain Trm. Specific Aim 2. Determine if DR neuronal signaling regulates CD8 T cell-mediated lymphocytic choriomeningitis virus (LCMV) induced VE or experimental cerebral malaria (ECM).

神经疾病是死亡和残疾的主要原因，尽管病因多样化，但 免疫功能障碍的常见主题。免疫系统和神经系统都经过细微调整 感官并策划对外部刺激的反应。大脑中有数十亿个神经元，但我们知道 几乎没有关于脑神经元如何与中枢神经系统中的免疫系统接口。最近的进步 神经免疫学包括发现脑膜淋巴的，免疫细胞在CNS稳态中的作用 以及CNS感染后大脑中持续存在的组织驻留记忆T细胞（TRM）的表征。我们 最近（Urban等，《自然免疫学》，2020年）提供了证据，表明TRM也是独特的 周围免疫和感染后的大脑，表明大脑独有的过程可能 外周免疫后，该器官中的TRM持久性。但是，未知是否特定神经元 大脑的途径有助于TRM的产生或持久性，以及神经元信号传导是否有助于 影响CNS TRM的保护。 人类中的几种中枢神经系统疾病具有免疫学病因，T细胞认为有助于 病毒性脑炎（VE）的发病机理，脑疟疾（CM）因此，第二个未知是脑的作用 T细胞介导的中枢神经系统疾病中的神经元信号通路。 我们将使用化学遗传学攻击这些问题。具体而言，有高度保守的神经元 在脑干中，例如背raphe（DR），该脑干在整个中枢神经系统中广泛投射并激活 对与免疫相关的信号的反应，例如压力，疼痛和瘙痒。表达Sert-的转基因B6小鼠 CRE目标博士神经元。表达Flox/Stop Designer的腺相关病毒的立体定位注射 专门由设计师药物（Dreadd）激活的受体，仅在Cre-中表达Dreadd 表达神经元。通过MCHERRY表达检测到表达CRE的神经元转导的神经元，可以是 根据其性质（由精确的Dreadd指定的激活或抑制信号）操纵，通过 注射设计师配体。我们已经获得了这些小鼠和病毒系统，并提供了我们能力的证明 进行复杂的立体定位手术，并将Dreadd构建体传递给靶向神经元。 我们将使用化学遗传技术和我们的能力来评估中枢神经系统免疫力和免疫介导 发病机理测试中心假设，即脑神经元信号通路有助于 CNS TRM的生成，维护和功能以及T细胞介导的CNS疾病的结果。 特定目的1。确定神经元信号是否调节发电，维护或保护性 脑TRM的功能。 特定目标2。确定神经元信号传导是否调节CD8 T细胞介导的淋巴细胞 绒毛膜炎病毒（LCMV）诱导VE或实验性脑疟疾（ECM）。