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

为了成功地感染目标宿主细胞，艾滋病毒必须执行感染的早期步骤，以整合到 宿主染色质。整合后，HIV完成了感染的后期步骤，在那里新的颗粒组装在一起。 这些病毒粒子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。许多实验室使用的多种方法已经开始 促进我们对不同步骤的动力学、动力学和细胞内定位的理解 早期事件包括病毒与细胞辅助因子或细胞先天/固有免疫因子的相互作用。 虽然这些系统都很有洞察力，但它们都有各自的优点和缺点，从而导致数据冲突 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衣壳是否保护了逆转录的基因组不受先天传感器的影响 核转位，例如cGAS。到目前为止，尚不清楚先天感知机制是如何针对病毒的。 颗粒以及这些颗粒是否被降解、在特定的细胞隔间积聚或感染 手机。在我的研究生涯中，我优化了研究HIV-1早期步骤的方法 来自HIV-2的分离物和各种循环中的SIV。我的长期职业计划是阐明各种病毒 机制以及它们与细胞的相互作用，利用我之前开发的技术。这些 方法监控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-- 并明确地将病毒行为与原代细胞的生殖性细胞感染联系起来。这 一项研究项目旨在阐明先天感知的参与者是否对传染性有直接影响 颗粒，或导致先天感知通路激活过程中的流产感染。的研究。 这些细胞和病毒机制将为可能被利用来 关于艾滋病毒/艾滋病流行的积极成果和应用。

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.