Imaging Virological Synapses During Parenteral HIV Transmission

描述（由申请人提供）：为了制定阻断HIV胃肠外传播的策略，了解启动HIV感染和病毒体内传播的机制至关重要。尽管25年来，该领域主要集中在病毒传播的无细胞机制上，以建模和了解该疾病，但可以通过感染细胞和未感染细胞之间的直接接触来证明艾滋病毒的传播是令人信服的。我们最近的研究集中在细胞间粘附结构的细胞生物学上，称为病毒学突触，它增强了细胞间的病毒传播。使用一种创新的策略来可视化病毒通过突触的传播，我们设计了一种荧光的、感染性的HIV分子克隆。在病毒学突触中，我们发现了受感染的T细胞中组装病毒颗粒的动态运动，以及伴随突触形成和感染的新型内吞过程。细胞过程的复杂性及其高效率表明，这可能是艾滋病毒传播的主要模式。我们将检验病毒学突触代表体内传播的基本单位的假设。用抗体或其他拮抗剂阻断突触可能是打破传递循环的关键。对于这个前卫的项目，我们将利用最近开发的小型人源化动物模型来检查在肠胃外HIV传播过程中发生的事件的顺序。我们将测试无细胞途径与细胞相关途径的相对效率，并在病毒攻击后的最初几分钟和几小时内跟踪病毒传播。为了在生物体水平上揭示细胞-细胞和细胞-病毒的相互作用，我们将采用体内成像方法来跟踪感染或携带病毒的细胞在人源化小鼠体内的运动。我们将揭示感染细胞/病毒和未感染细胞之间的相互作用序列。这一结果将为干扰病毒性T细胞提供新的策略。

DESCRIPTION (provided by applicant): 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 t