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.

为了成功感染靶宿主细胞，HIV必须执行感染的早期步骤，以整合到靶细胞中。 宿主染色质。整合后，HIV完成了感染的最后步骤，新的粒子聚集在一起。 这些病毒体含有病毒宿主蛋白和RNA病毒基因组。最近，艾滋病毒感染的早期步骤 都受到了严格的审查一些实验室采用的多种方法已经开始 推进我们的动力学，动力学，和细胞内的位置的不同步骤的理解， 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。 尽管这些系统都很有见地，但它们都有优点和缺点，导致数据冲突， 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 争论的焦点是，HIV衣壳是否能保护逆转录的基因组不受先天传感器的影响， 核易位，例如cGAS。到目前为止，尚不清楚先天感觉机制如何靶向病毒， 颗粒以及这些颗粒是否被降解，在特定的细胞区室中积累，或感染 cell.在我的整个研究生涯中，我优化了研究HIV-1早期阶段的方法，主要是 HIV-2和各种SIV的分离株。我的长期职业规划是阐明各种病毒 机制及其与细胞的相互作用，通过利用我以前开发的技术。这些 方法监测单个病毒颗粒的行为，并直接将颗粒行为与成功的 细胞感染该提案的主要目标是澄清以下方面的早期步骤的关键方面： HIV感染，并明确地将病毒行为与原代细胞的生产性细胞感染联系起来。这 一项研究项目旨在澄清先天感觉中的参与者是否对传染病有直接影响。 颗粒，或导致先天感应途径激活期间的流产感染。研究 这些细胞和病毒机制将为细胞机制提供新的线索， 在艾滋病毒/艾滋病流行病方面取得了积极成果和应用。