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: 10188906
• 负责人: Joshua Woodward
• 金额: $ 25.05万
• 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 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188906
• 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.

项目概要 C-di-GMP、c-di-AMP 和 3′3′-cGAMP 等是调节多种细菌的第二信使 引起宿主免疫的细菌生理学和病原体相关分子模式（PAMP）的各个方面 这些 CDN 广泛由细菌产生，更重要的是，对于感染过程中的反应至关重要。 人类病原体的发病机制，包括 B 族链球菌、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菌也有显着的CDN。 由于其能够增强 I 型干扰素和适应性，因此有潜力作为传染病的疫苗佐剂 识别 PAMP 的模式识别受体 (PRR) 是免疫反应的第一道防线。 因此，CDN PRR 的鉴定和表征对于细菌感染的研究至关重要。 传染病的发病机制和 CDN 作为疫苗佐剂的应用在这里，我们鉴定了 Kvb2， 电压依赖性钾通道 Kv1 的胞质 b 亚基，作为 c-di-AMP 相互作用蛋白 c-di-AMP 亲和力下拉测定 Kvb2 是一种功能性醛酮还原酶 (AKR)，可调节细胞。 兴奋性取决于其 NADPH 辅因子 Kvβ2 的氧化和氧化还原状态，广泛表达于体内。 然而，Kvb2 功能障碍的主要后果是导致人类大脑、心脏和淋巴器官。 我们发现 Kvb2 是一种先天性神经损伤，会导致记忆障碍和癫痫发作。 神经免疫系统中的免疫卫士，通过检测 CDN 和电位来感知细菌感染 我们的目标是 (i) 表征 Kvb2 对 CDN 结合的特异性和动态，以及 Kvb2 对 CDN 结合的反应调节 Kv 通道的机制；(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反应。 我们的研究将细胞的兴奋性与检测结合起来。 细菌感染，这将拓宽我们对 CDN 介导的中枢抗菌免疫的理解 神经系统。