Direct live-cell visualization and quantification of interactions between infectious HIV particles in primary target cells.

主要靶细胞中感染性 HIV 颗粒之间相互作用的直接活细胞可视化和量化。

• 批准号: 9695810
• 负责人: Joao Filipe Inacio Mamede
• 金额: $ 16.2万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-06 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9695810
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Behavior; Behavior monitoring; Binding Proteins; Biological Assay; Blood; CD4 Positive T Lymphocytes; Calorimetry; Capsid; Cell model; Cells; Chromatin; Complex; Conflict (Psychology); Cyclic GMP; Cytoplasm; DNA; Data; Dendritic Cells; Detection; Development Plans; Environment; Epidemic; Etiology; Event; Genome; Goals; HIV; HIV Infections; HIV-1; HIV-2; Imaging Techniques; Immunologic Factors; Individual; Infection; Kinetics; Laboratories; Lead; Light; Location; Mentors; Methods; Microtubules; Molecular; Myelogenous; Nuclear Pore Complex; Nuclear Translocation; Nucleic Acids; Outcome; Pathogenesis; Pathway interactions; Population Analysis; Primary Infection; Protein Subunits; Proteins; Reporting; Research; Research Project Grants; Reverse Transcription; SIV; Series; Site; Stimulator of Interferon Genes; Structure; Suggestion; Surface Plasmon Resonance; System; Techniques; Titrations; Viral; Viral Genome; Viral Proteins; Virion; Virus; Virus Integration; Virus Replication; Visualization; analytical method; antiretroviral therapy; biophysical properties; biophysical techniques; career; career development; imaging modality; insight; knowledge of results; live cell imaging; macrophage; novel strategies; pandemic disease; particle; polyglutamine; response; scale up; sensor; trafficking; viral RNA

To successfully infect a target host-cell HIV has to perform the early-steps of infection to integrate into the host chromatin. After integration, HIV completes the late-steps of infection where new particles assemble. These virions contain viral host proteins and the RNA viral genome. Recently, the early steps of HIV infection have been under intense scrutiny. Multiple approaches utilized by a number of laboratories have begun to advance our understanding of the kinetics, dynamics, and intracellular location of the different steps of the early events including viral interaction with cellular co-factors, or cellular innate/intrinsic immune factors. Although insightful, each of these systems have strengths and weaknesses leading to conflicting data due to different cellular models, techniques, and analysis of populations of viruses that do not infect a cell. A question under debate is whether the HIV capsid shields the reverse transcribed genome from innate sensors before nuclear translocation, e.g. cGAS. To date, it is not clear how the innate sensing machinery targets viral particles and whether these particles are degraded, accumulate in specific cellular compartments, or infect a cell. Throughout my research career, I have optimized methods that study the early-steps of HIV-1, primary isolates from HIV-2, and various circulating SIVs. My long-term career plan is to elucidate the various viral mechanisms and their interactions with the cell by utilizing techniques that I previously developed. These methods monitor the behavior of individual viral particles and directly connect particle behavior to successful cellular infection. The main goal of this proposal is the clarification of key aspects of the early-steps of HIV infection and to definitively connect viral behavior to productive cell infection of primary cells. This research project aims to clarify if the actors in innate sensing have a direct impact on infectious particles, or lead to an abortive infection during the activation of innate sensing pathways. The study of these cellular and viral mechanisms will shed new light into cellular mechanisms that could be utilized to have a positive outcome and application on the HIV/AIDS epidemic.

为了成功感染目标宿主细胞，HIV 必须执行感染的早期步骤，以整合到目标宿主细胞中。 宿主染色质。整合后，艾滋病毒完成了感染的后期步骤，新的颗粒聚集在一起。 这些病毒颗粒含有病毒宿主蛋白和 RNA 病毒基因组。最近，HIV感染的早期步骤 都受到严格审查。许多实验室已开始采用多种方法 增进我们对不同步骤的动力学、动力学和细胞内位置的理解 早期事件包括病毒与细胞辅助因子或细胞先天/内在免疫因子的相互作用。 尽管很有洞察力，但这些系统都有优点和缺点，导致数据冲突，因为 不同的细胞模型、技术和对不感染细胞的病毒群体的分析。一个问题 目前争论的焦点是，HIV 衣壳是否能保护逆转录基因组免受先天传感器的影响？ 核易位，例如气体。迄今为止，尚不清楚先天传感机制如何针对病毒 颗粒以及这些颗粒是否被降解、积聚在特定的细胞区室中或感染 细胞。在我的研究生涯中，我优化了研究 HIV-1 早期步骤的方法，主要是 从 HIV-2 和各种循环的 SIV 中分离出来。我的长期职业计划是阐明各种病毒式传播 通过利用我之前开发的技术来了解它们的机制及其与细胞的相互作用。这些 方法监测单个病毒颗粒的行为并将颗粒行为与成功直接联系起来 细胞感染。该提案的主要目标是澄清早期步骤的关键方面 HIV 感染并明确将病毒行为与原代细胞的生产性细胞感染联系起来。这 研究项目旨在澄清先天感知的参与者是否对传染性有直接影响 颗粒，或在先天传感通路激活期间导致流产感染。研究的 这些细胞和病毒机制将为细胞机制提供新的线索，可用于 在艾滋病毒/艾滋病流行方面取得了积极的成果和应用。