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， 了解启动HIV感染和病毒在体内传播的机制。 虽然25年来，该领域主要集中在无细胞机制， 病毒传播的模型和了解疾病，一个令人信服的情况下， 艾滋病病毒通过感染者和病人之间的直接接触传播， 未感染的细胞我们最近的研究主要集中在 细胞间的粘附结构，称为病毒学突触， 在细胞间传播。使用创新策略可视化病毒传播 通过突触，我们设计了一个荧光，传染性的分子克隆， 艾滋病。在病毒学突触上，我们发现了 感染T细胞中的病毒颗粒，以及一种新的内吞过程， 伴随着突触的形成和感染。细胞的复杂性 过程及其高效率表明，这可能是主导的 艾滋病毒传播的范例。我们将检验病毒学 突触代表体内传输的基本单位。阻断 突触，无论是与抗体或与其他拮抗剂，可能是关键， 打破传播的循环。为了这个前卫的项目，我们将利用 最近开发的小型人源化动物模型来检查 发生在非肠道HIV传播过程中的事件。我们将测试相对 无细胞途径与细胞相关途径的效率，并跟踪病毒传播 在病毒攻击后的最初几分钟和几小时内。为了揭示细胞-细胞， 在生物体水平上的细胞-病毒相互作用，我们将采用体内成像 追踪受感染或携带病毒的细胞在体内运动的方法 人源化小鼠。我们将揭示受感染者与 细胞/病毒和未感染的细胞。研究结果将提供新的战略， 干扰病毒传播和扩散。

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.