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

基本信息

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
项目类别：
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

项目摘要

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).

神经系统疾病是死亡和残疾的主要原因，尽管病因多种多样，但都有一个共同点免疫功能障碍的共同主题。免疫系统和神经系统都针对这两者进行了微调感知并协调对外部刺激的反应。大脑中有数十亿个神经元，但我们知道关于大脑神经元如何与中枢神经系统中的免疫系统相互作用几乎一无所知。最近的进展神经免疫学包括脑膜淋巴管的发现、免疫细胞在中枢神经系统稳态中的作用以及中枢神经系统感染后大脑中持续存在的组织驻留记忆 T 细胞 (Trm) 的特征。我们最近（Urban 等人，自然免疫学，2020）提供的证据表明 Trm 也是在外周免疫和感染后的大脑，表明大脑独特的过程可能外周免疫后促进 Trm 在该器官中的持续存在。然而，尚不清楚是否有特定的神经元大脑中的通路有助于该器官中 Trm 的产生或持续存在，以及神经元信号传导是否影响 CNS Trm 的保护。人类的几种中枢神经系统疾病都有免疫学病因，T 细胞被认为有助于病毒性脑炎（VE）、脑型疟疾（CM）的发病机制因此，第二个未知数是大脑的作用 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来解决这些问题。具体来说，存在高度保守的神经元脑干内，例如中缝背侧 (DR)，其广泛投射到整个中枢神经系统并在对压力、疼痛和瘙痒等免疫相关信号的反应。表达SERT-的转基因B6小鼠 cre 靶向 DR 神经元。表达 flox/stop 设计器的腺相关病毒的立体定位注射受体专门由设计药物（DREADD）激活，导致 DREADD 仅在创造中表达表达神经元。通过 mCherry 表达检测转导的 cre 表达神经元，并且可以根据其性质（由精确的 DREADD 指定的激活或抑制信号）进行操纵注射设计配体。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执行复杂的立体定位手术并将 DREADD 结构传递到目标神经元。我们将利用化学遗传学技术和我们的能力来评估中枢神经系统免疫和免疫介导的发病机制来检验大脑神经元信号通路有助于 CNS Trm 的产生、维持和功能以及 T 细胞介导的 CNS 疾病的结果。具体目标 1. 确定 DR 神经元信号传导是否调节生成、维持或保护大脑 Trm 的功能具体目标 2. 确定 DR 神经元信号传导是否调节 CD8 T 细胞介导的淋巴细胞脉络膜脑膜炎病毒 (LCMV) 诱发 VE 或实验性脑型疟疾 (ECM)。