Kvbeta2 and the host response to cyclic dinucleotides

Kvbeta2 和宿主对环状二核苷酸的反应

• 批准号: 10358622
• 负责人: Joshua Woodward
• 金额: $ 20.59万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-23 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10358622
• 关键词: Affinity; Anti-Bacterial Agents; Bacteria; Bacterial Infections; Bacterial Physiology; Binding; Biochemical; Biological Assay; Blood capillaries; Brain; Breeding; Cell Line; Cells; Chlamydia trachomatis; Communicable Diseases; Complement; Complement Receptor; Complex; Coupled; Detection; Dinucleoside Phosphates; Eukaryotic Cell; Exhibits; Family; Family member; Foundations; Functional disorder; Future; Germ Lines; Growth; Heart; Human; Immune; Immune response; Immunity; Impairment; In Vitro; Infection; Inflammasome; Inflammation Mediators; Inflammatory; Inflammatory Response; Innate Immune Response; Interferons; Knockout Mice; Ligands; Listeria monocytogenes; Mediating; Membrane; Memory; Memory impairment; Meningitis; Microglia; Modeling; Molecular; Mus; Mutagenesis; Mycobacterium tuberculosis; NADP; Natural Immunity; Neuraxis; Neurodegenerative Disorders; Neuroimmune system; Neurologic; Nucleotides; Oxidation-Reduction; Oxidative Stress; Oxidoreductase; Pathogenesis; Pattern; Pattern recognition receptor; Periodicity; Potassium; Potassium Channel; Production; Proteins; Proteomics; Radial; Reactive Oxygen Species; Reporting; Role; Second Messenger Systems; Seizures; Signal Transduction; Specificity; Staphylococcus aureus; Streptococcus Group B; Stress; Testing; Vaccine Adjuvant; Virus Diseases; Work; adaptive immune response; base; cofactor; cytokine; human pathogen; immunogenicity; insight; lymphoid organ; microorganism; mouse model; novel; pathogen; pathogenic bacteria; receptor; relating to nervous system; response; voltage

PROJECT SUMMARY C-di-GMP, c-di-AMP, and 3′3′-cGAMP, among others, are bacterial second messengers that regulate a variety aspects of bacterial physiology and pathogen-associated molecular patterns (PAMPs) that elicit host immune responses during infection. These CDNs are widely produced by bacteria, and more importantly, crucial for the pathogenesis of human pathogens including Group B Streptococcus, Mycobacterium tuberculosis, Staphylococcus aureus, Chlamydia trachomatis and Listeria monocytogenes. CDNs also have significant potential as vaccine adjuvants for infectious diseases due to their capability to boost type I IFN and adaptive immune responses. Pattern recognition receptors (PRRs) that recognize PAMPs are the first line defense for bacterial infection. Identification and characterization of CDN PRRs are therefore critical to the study of pathogenesis of infectious diseases and the application of CDNs as vaccine adjuvants. Here, we identified Kvb2, the cytosolic b subunit of voltage-dependent potassium channel Kv1, as a c-di-AMP-interacting protein through c-di-AMP affinity pull-down assay. Kvb2 is a functional aldo-keto reductase (AKR) which modulates the cellular excitability depending on the oxidative and redox status of its NADPH cofactor. Kvβ2 is widely expressed in human brain, heart and lymphoid organs. However, the dominating consequence of Kvb2 dysfunction is causing neurological impairments that leads to memory impairments and seizures. We hypothesize that Kvb2 is an innate immune guard in the neuroimmune system that senses bacterial infection by detecting CDNs and potential oxidative stresses. We aim to (i) characterize the specificity and dynamics of Kvb2 for CDN binding, as well as the mechanisms of Kv channel modulation by Kvb2 response to CDN binding; (ii) investigate the role of Kvb2 in restricting bacterial growth and modulating innate immune responses using a microglia cell line and L. monocytogenes meningitis mouse model. Our studies will couple the excitability of cells with detection of bacterial infection, which will broaden our understanding of CDN-mediated antibacterial immunity in the central nervous system.

项目摘要 C-DI-GMP，C-DI-AMP和3'3'-CGAMP等是细菌第二使者，可调节多样性 细菌生理学和病原体相关的分子模式（PAMP）的各个方面引起宿主免疫 感染过程中的反应。这些CDN被细菌广泛产生，更重要的是，对于 人类病原体的发病机制，包括B组链球菌，结核分枝杆菌， 金黄色葡萄球菌，沙眼衣原体和单核细胞增生李斯特氏菌。 CDN也有明显的 由于其能力提高I型和自适应的能力，因此作为感染疾病的疫苗调节剂的潜力 免疫反应。识别弹药的模式识别接收器（PRR）是第一行的防御 细菌感染。因此，CDN PRR的识别和表征对于研究至关重要 传染病的发病机理以及CDN作为疫苗调节剂的应用。在这里，我们确定了KVB2， 电压依赖性钾通道KV1的胞质B亚基，作为C-DI-PAMP相互作用蛋白通过 C-DI-AMP亲和力下拉测定法。 KVB2是一种功能性的Aldo-Keto降低酶（AKR），可调节细胞 兴奋性取决于其NADPH辅因子的氧化和氧化还原状态。 KVβ2在 人脑，心脏和淋巴器官。但是，KVB2功能障碍的主导后果正在引起 导致记忆力障碍和癫痫发作的神经障碍。我们假设KVB2是天生的 神经免疫系统中的免疫卫队通过检测CDN和电势来感知细菌感染 氧化应激。我们的目标是（i）表征KVB2对CDN结合的特异性和动力学，以及 KVB2对CDN结合的反应KV通道调制的机制； （ii）研究KVB2在 使用小胶质细胞系和L。限制细菌生长并调节先天免疫复杂。 单核细胞增生脑膜炎小鼠模型。我们的研究将使细胞的兴奋与检测 细菌感染将扩大我们对中央CDN介导的抗菌免疫的理解 神经系统。