Complement Resistance Acquired During Acute to Persistent Rubulavirus Infection

急性至持续性风疹病毒感染期间获得的补体耐药性

• 批准号: 10645486
• 负责人: Griffith D. Parks
• 金额: $ 24.12万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-23 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10645486
• 关键词: Acute; Air; Binding; Biochemical; Biological Assay; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Clinical; Clinical Trials; Complement; Complement Factor H; Complement Inactivators; Cytolysis; Data; Drug Modulation; Engineering; Environment; Epithelial Cells; Fibrinogen; Future; Genetic Polymorphism; Genome; Goals; Human; In Vitro; Infection; Inflammation; Innate Immune System; Liquid substance; Lung; Lung infections; Mediating; Mumps virus; Paramyxovirus; Pathway interactions; Population; Production; Proteomics; RNA Virus Infections; Resistance; Respiratory System; Respiratory Tract Infections; Role; Rubulavirus; Rubulavirus Infections; Severities; Surface; Testing; Therapeutic; Time; Trachea; Variant; Viral; Viral Respiratory Tract Infection; Virion; Virus; Virus Diseases; Virus Replication; Vitronectin; Work; acute infection; chronic infection; conditioning; gene complementation; inhibitor; innate immune mechanisms; innate immune pathways; interest; parainfluenza virus; pathogen; prototype; recruit; respiratory; respiratory infection virus; response; sulfated glycoprotein 2; transcriptomics

For many RNA virus infections, an initial acute infection can transition to a prolonged or persistent infection, in which infected cells survive and continue to produce progeny virus. Complement (C’) is a powerful innate immune system which can directly lyse virus-infected cells or neutralize virus (1-4), but the role of C’ in controlling persistent respiratory RNA virus infections is not well understood. Given that viruses have mechanisms to block C’-mediated cell lysis, persistent infections can set up a prolonged inflammation cycle – where activated C’ continues to provide damaging inflammation, but viral inhibitory mechanisms block elimination of pathogen and infected cells. This project emerged from our striking findings that during an initial acute infection of human lung cells with the Rubulavirus Parainfluenza virus 5 (PIV5), infected cells are very sensitive to C’-mediated lysis. Importantly however, after transitioning to a persistent infection, PIV5-infected cells are nearly completely resistant to C’ lysis. Our transcriptomics data show that PIV5 acutely infected cells have low level expression of C’ inhibitors, but this shifts to high level expression of cellular C’ inhibitors Factor H, Factor I, Vitronectin and Clusterin in persistently infected cells. Our central hypothesis is that PIV5 persistently infected cells acquire resistance to C’-mediated lysis due to their acquired ability to express high levels of C’ inhibitors Factor H and Vitronectin. Our goals are to identify: 1) the mechanisms for acquiring C’ resistance during the PIV5 acute-to-persistent transition (Aim 1), and 2) consequences of this shift for production of C’-resistant virus (Aim 2). Aim 1 will define the mechanism for differential sensitivity of airway cells to C’-mediated lysis during transition from acute to persistent infection. Engineered respiratory tract cell lines and primary tracheal or bronchial air-liquid interface (ALI) cell cultures will be used to test the hypothesis that synthesis of C’ inhibitors Factor H and Vitronectin by persistently infected cells results in conditioning of the cells to be resistant to C’-mediated lysis. Aim 2 will identify C’ factors associated with virus particles derived from acute versus persistently infected cells and define the sensitivity of persistent virus to C’- mediated neutralization. Proteomics and biochemical assays will test the hypothesis that virus derived from persistently infected cells will be C’-resistant due to recruitment of Factor H or Vitronectin. Results from our work on C’ interactions with persistent RNA virus infections will have strong potential to inform therapeutics, given: 1) the clinical impact of prolonged viral respiratory infections, 2) polymorphisms in C’ genes can correlate with severity of viral infections, and 3) clinical trials for respiratory tract infections are underway with drugs that modulate C’ responses.

对于许多RNA病毒感染，初始急性感染可以过渡到持续的 感染，其中感染细胞生存并继续产生进展的病毒。补充（C'）是 强大的先天免疫抑制系统，可以直接裂解病毒感染的细胞或中和病毒（1-4），但 C’在控制持续的呼吸道RNA病毒感染中的作用尚不清楚。鉴于 病毒具有阻断C'-介导的细胞裂解的机制，持续感染可以延长 炎症周期 - 激活C'继续提供损害感染，但病毒抑制 机制阻止了消除病原体和感染细胞的消除。 该项目来自我们的打击发现，在最初的急性感染人类肺部 具有rubulavirus parainfluenza病毒5（PIV5）的细胞，感染的细胞对C'M介导的细胞非常敏感 裂解。但是，重要的是，过渡到持续感染后，PIV5感染的细胞几乎是 完全抵抗C的裂解。我们的转录组学数据表明，PIV5急性感染的细胞低 C'抑制剂的水平表达，但这转移到细胞C'抑制剂因子H的高水平表达， 因子I，玻璃体蛋白和簇蛋白持续感染细胞。 我们的中心假设是PIV5持续感染的细胞获得了对C'-介导的抗性 由于其获得的能力表达高水平的C抑制剂因子H和玻璃体蛋白而引起的裂解。我们的 目标是识别：1）在PIV5急性之间获得C抗性的机制 过渡（AIM 1）和2）这种转变对C'-耐药病毒的产生（AIM 2）。 AIM 1将定义气道细胞对C介导的裂解的差异敏感性的机制 从急性到持续感染的过渡期间。设计的呼吸道细胞系和主要 气管或支气管空气界面（ALI）细胞培养物将用于检验以下假设。 通过持续感染的细胞合成C抑制剂因子H和玻璃体蛋白会导致调节 细胞对C介导的裂解具有抗性。 AIM 2将识别与病毒颗粒相关的C因子 源自急性与持续感染的细胞，并定义持续病毒对C'-的敏感性 介导的神经化。蛋白质组学和生化测定将检验病毒衍生的假设 由于因子H或玻璃纤维素的募集，因此持续感染的细胞将具有C'耐药性。 我们在C与持续RNA病毒感染的C互动的工作的结果将具有很强的 给予治疗的潜力：1）长期病毒呼吸道感染的临床影响，2） C的基因中的多态性可能与病毒感染的严重程度相关，3）临床试验的临床试验 呼吸道感染正在进行调节C反应的药物。