Project 1: Dissecting Persistent Virus Reservoirs in Tissues

项目 1：剖析组织中的持久病毒库

• 批准号: 10666579
• 负责人: Thomas Hope
• 金额: $ 23.77万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666579
• 关键词: Ablation; Anatomy; Animals; Antibodies; Aquila; Area; Autopsy; Binding; Biological Models; CD4 Positive T Lymphocytes; Cells; Cellular Morphology; Collaborations; DNA; Dendritic Cells; Detection; Down-Regulation; Environment; Evaluation; Grant; HIV; Image; Immobilization; Infection; Inflammation; Interruption; Intervention; Investigation; Kinetics; Knowledge; Label; Location; Lymphoid Tissue; Macaca mulatta; Macrophage; Methods; Modeling; Monitor; Mucous Membrane; Myeloid Cells; Names; Nature; Nucleic Acids; PET/CT scan; Phase; Physiology; Play; Population; Population Dynamics; Positron-Emission Tomography; Process; Production; Proteins; RNA; Radioactive; Reproducibility; Resolution; Role; SIV; Sampling; Scanning; Signal Transduction; Site; Source; T-Lymphocyte; Techniques; Technology; Time; Tissue Banks; Tissue Sample; Tissues; Transmission Electron Microscopy; Trauma; Viral Antigens; Viral Proteins; Viral reservoir; Viremia; Virion; Virus; Visualization; acute infection; antiretroviral therapy; innovation; insight; mast cell; nonhuman primate; pathogen; prevent; programs; recruit; simian immunodeficiency virus gp120; therapy development; tissue repair; viral rebound

PROJECT SUMMARY/ABSTRACT Without a complete knowledge of the formation of the HIV reservoir, its persistence, and potential rebound, it will be difficult to develop therapies to achieve a cure for HIV. A major limitation that the HIV field needs to overcome in order to advance our understanding of viral rebound is the inability to systemically study the reservoir in tissues. With the support of a P01 focused on viral rebound of the early reservoir, we have recently optimized a new workflow that we named PET-CT guided necropsy. This workflow is based on the use of a 64Cu-labeled anti-SIV env FAB2 (64Cu-7D3FAB2) to guide the sampling and/or necropsy process to identify and collect pieces of tissues containing foci of 64Cu signal revealing sites of SIV replication. The presence of virus infected cells in these “hot” tissues is now validated by 4 markers (Gag and Env expression and CD4 and MHC-I downregulation). Unexpectedly, we only detect SIVmac239 infection of myeloid cells during the first 7-10 days after ATI. No SIV infected T cells are observed although CD4 T cells are abundant in these foci. Finally, our 64Cu-7D3FAB2 probe led us to tissues where we can reproducibly visualize virion producing cells, with a myeloid cell morphology, by transmission electron microscopy. This multiscaling imaging, leveraging PET/CT guided necropsy, allows us to peak into the elusive tissue reservoir. In this Project 1 of the Program, we propose to leverage our advancements with PET-CT-guided sampling of tissues as well as additional innovative sampling techniques in non-human primate (NHP) models to dissect the dynamics of the persistent SIV reservoir. Specifically, we have planned 3 different type of NHP studies that, with the help of ad-hoc interventions, will give us an unprecedented resolution into the dynamics of the reservoir and its rebound. We have planned interventions that aim at “tickling” and disturbing the reservoir during cART, which will allow PET-CT-signal increase and collection of tissue samples that will be analyzed with different techniques including multiscale imaging and distributed to the other 2 projects. We will “mulch” the reservoir by favoring its seeding into mucosal tissues with DSS treatment before infection and then again an in-depth investigation of the tissue reservoir at the time of cART initiation and after several months of suppressive cART. Finally, we will deplete or stimulate myeloid cells, which our studies have revealed as main contributor to viral rebound after ATI and study the impact of these interventions on the reservoir dynamics and timing of rebound viremia. In summary, our project will use different strategies in NHP studies to clarify the nature and source of the SIV reservoir with particular focus at the mucosal tissue and myeloid cells.

项目摘要/摘要 如果不完全了解艾滋病毒蓄水池的形成、其持久性和潜在的反弹，它将 很难开发出治愈艾滋病毒的疗法。艾滋病毒领域需要克服的一个主要限制 为了增进我们对病毒反弹的理解，我们无法系统地研究 纸巾。在关注早期病毒式反弹的P01的支持下，我们最近优化了一个 我们命名为PET-CT引导尸检的新工作流程。此工作流程基于64Cu型标签的使用 抗SIV env FAB2(64Cu-7D3FAB2)，用于指导采样和/或尸检过程，以识别和收集碎片 含有64Cu病灶的组织的信号揭示了SIV复制的位置。病毒感染细胞在体内的存在 这些“热”组织现在被4个标记物证实(GAG和Env的表达以及CD4和MHC-I的下调)。 出乎意料的是，我们只在ATI后的头7-10天检测到SIVmac239感染髓系细胞。无SIV 虽然这些病灶中有大量的CD4T细胞，但仍可观察到感染的T细胞。最后，我们的64Cu7D3FAB2探测器 将我们带到了组织中，在那里我们可以重复地看到产生病毒粒子的细胞，具有髓系细胞的形态，通过 透射电子显微镜。这种多尺度成像，利用PET/CT引导的尸检，使我们能够 进入难以捉摸的组织储藏室。在该计划的这个项目1中，我们建议利用我们的进步 利用PET-CT引导的组织采样以及其他创新的非人类采样技术 灵长类(NHP)模型来剖析持久性SIV储集层的动力学。具体地说，我们计划了3个 不同类型的国家惠普研究，在特别干预的帮助下，将为我们提供前所未有的解决方案 水库的动态和它的反弹。我们有计划的干预措施，目的是“挠痒痒”和 在CART期间扰乱水库，这将允许PET-CT信号增加和组织样本收集 这将用包括多尺度成像在内的不同技术进行分析，并分发给其他两个项目。 我们将通过在感染前用DSS治疗将其播种到粘膜组织中来覆盖水库。 然后再次对CART启动时的组织储存库进行深入调查 几个月的压抑大车。最后，我们将耗尽或刺激髓系细胞，我们的研究揭示了这一点 作为ATI后病毒反弹的主要贡献者，并研究这些干预措施对宿主的影响 反跳性病毒血症的动态和时间。总而言之，我们的项目将在NHP研究中使用不同的策略来 明确SIV储存库的性质和来源，特别是粘膜组织和髓样细胞。