Imaging Virological Synapses During Parenteral HIV Transmission

摘要： 为了制定阻断艾滋病毒非肠道传播的战略，至关重要的是 了解在体内引发艾滋病毒感染和病毒传播的机制。 尽管25年来，该领域主要关注的是无细胞机制 病毒传播来模拟和了解疾病，一个令人信服的案例可以 用于通过感染者和患者之间的直接接触传播艾滋病毒 未受感染的细胞。我们最近的研究集中在细胞生物学上。 细胞间黏附结构，称为病毒学突触，可增强病毒 在细胞间传播。使用创新策略将病毒传播可视化 通过突触，我们设计出了一种具有感染性的荧光分子克隆 爱滋病毒。在病毒学突触，我们发现了组装的动态运动 感染T细胞中的病毒颗粒，以及一种新的内吞过程，该过程 伴随突触形成和感染。细胞的复杂性 进程及其高效率表明这可能是占主导地位的 艾滋病毒传播的范例。我们将测试病毒学的假设 突触是体内信息传递的基本单位。阻挡 突触，无论是与抗体或与其他拮抗剂，可能是关键 打破了传播的循环。对于这个前卫的项目，我们将利用 最近开发的小型人性化动物模型来检查序列 在非肠道内传播艾滋病毒期间发生的事件。我们将测试相关的 无细胞途径与细胞相关途径的效率，并跟踪病毒传播 在病毒式挑战后的第一分钟和几个小时内。以显示细胞-细胞和 在生物体水平上的细胞-病毒相互作用，我们将使用活体成像 跟踪受感染或携带病毒的细胞在 人性化的老鼠。我们将揭示感染病毒之间的一系列相互作用 细胞/病毒和未感染细胞。研究结果将为以下方面提供新的战略 干扰病毒传播和传播。

ABSTRACT: To develop strategies to block parenteral transmission of HIV, it is critical to understand the mechanisms that initiate HIV infection and viral spread in vivo. Though for 25 years, the field has largely focused on cell-free mechanisms of viral spread to model and understand the disease, a compelling case can be made for the spread of HIV through direct contact between infected and uninfected cells. Our recent studies have focused on the cell biology of intercellular adhesive structures, called virological synapses, that enhance viral spread between cells. Using an innovative strategy to visualize viral transmission through synapses, we engineered a fluorescent, infectious molecular clone of HIV. At virological synapses, we uncovered dynamic movements of assembling virus particles in infected T cells, as well as a novel endocytic process that accompanies synapse formation and infection. The complexity of the cellular processes and their high efficiency suggests that this may be the dominant paradigm for HIV dissemination. We will test the hypothesis that the virological synapse represents the fundamental unit of transmission in vivo. Blocking the synapse, either with antibodies or with other antagonists, may be the key to breaking the cycle of transmission. For this avant garde project, we will exploit recently developed small humanized animal models to examine the sequence of events that occurs during parenteral HIV transmission. We will test the relative efficiency of cell-free versus cell-associated routes, and track viral dissemination in the first minutes and hours after viral challenge. To reveal the cell-cell and cell-virus interactions at an organismal level, we will employ in vivo imaging approaches to track the movement of infected or virus-carrying cells within humanized mice. We will reveal a sequence of interactions between infected cells/virus and uninfected cells. The results will provide new strategies to interfere with viral transmission and dissemination.