Defining a Role for Liver Myeloid Cells in Viral Persistence under ART

定义肝髓细胞在 ART 下病毒持续存在中的作用

• 批准号: 10654037
• 负责人: Mario Stevenson
• 金额: $ 51.04万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654037
• 关键词: Address; Adopted; Alleles; Anatomy; Antibodies; Area; Binding; Blood specimen; CD4 Positive T Lymphocytes; Cell Compartmentation; Cell membrane; Cells; Cellular Tropism; Characteristics; Clinical; Discrimination; Disease remission; Exhibits; Genetic Polymorphism; Genome; HIV; HIV-1; HIV/AIDS; Immune system; In Vitro; Individual; Infection; Interruption; Kupffer Cells; Length; Libraries; Ligands; Liver; Location; Macrophage; Membrane; Methods; Molecular; Molecular Cloning; Myelogenous; Myeloid Cells; Nature; Phenotype; Phylogenetic Analysis; Plasma; Plasma Cells; Polymerase; Proviruses; Recombinants; Research; Resources; Role; Sampling; Structure; Tissues; Transplantation; Variant; Viral; Viral reservoir; Viremia; Virion; Virus; antiretroviral therapy; design; insight; molecular clock; novel; novel strategies; personalized approach; viral rebound

The pursuit of a cure for HIV-1 infection is founded on the hypothesis that reservoirs that support viral persistence are amenable to elimination. As such, most of the research on HIV-1 cure and remission has centered on identifying the dynamics of CD4+ T cell reservoirs as well as on approaches to eliminate them. We recently presented evidence that following analytic treatment interruption (ATI), rebounding viremia comprised virions that were highly macrophage tropic. Through the use of a novel immunoaffinity enrichment method, we further demonstrated that macrophage-tropic viruses in post-ATI plasma have a myeloid cell origin [1]. These observations define myeloid cells as a reservoir under suppressive ART and that these reservoirs require consideration in the design of viral remission and cure strategies The myeloid compartments that house HIV-1 reservoirs under ART are unkown. However, the approaches we developed have the capacity to yield this information. Those approaches hinge upon our demonstration that virion membranes are derived from the host cell plasma membrane and as such, virions harbor ligands that inform on their host cell of origin. We will adopt these approaches to address the long-standing issue of whether Kupffer Cells in the liver, which is the largest myeloid compartment of the immune system, serve as viral reservoirs in the face of suppressive ART. Our objectives are: Aim1: Identify host cell ligands on virions derived from primary Kupffer cells that enable immunoaffinity isolation of virions with a Kupffer cell origin in post ATI-plasma. Aim 2: Derive libraries of full length viral envelopes from liver tissue of HIV-1-infected individuals and assess their cellular tropism and dynamics by high-throughput viral phenotyping and Molecular Clock analysis, respectively. Aim 3: Employ liganded virion capture and high discrimation Taq (HiDi Taq) polymerase-directed allele amplification to assess whether virions in tissue-matched plasma originate from Kupffer cells and whether immunocaptured virions from plasma and proviruses from liver tissue exhibit related virological characteristics and phylogenetic structures. This proposal leverages important clinical resources that will be used to define whether Kupffer cells serve as viral reservoirs of HIV-1. The information derived from this proposal will provide the rationale for the identification of tailored approaches to eliminate myeloid reservoirs within the liver and other myeloid compartments.

追求HIV-1感染的治疗方法是基于支持病毒持久性的储层的假设 可以消除。因此，关于HIV-1治愈和缓解的大多数研究都集中在 识别CD4+ T细胞库的动力学以及消除它们的方法。我们最近 提出的证据表明，在分析治疗中断（ATI）之后，反弹的病毒率包括病毒体 是高度巨噬细胞的热带。通过使用一种新型免疫亲和力富集方法，我们进一步 证明ATI后血浆中的巨噬细胞 - 循环病毒具有髓样细胞的起源[1]。这些 观察结果将髓样细胞定义为抑制性艺术的储层，这些储层需要 考虑病毒缓解和治愈策略的设计 在艺术中​​容纳HIV-1水库的髓样室是不可行的。但是，我们的方法 开发有能力产生此信息。这些方法取决于我们的演示 病毒体膜源自宿主细胞质膜，因此，病毒体具有配体 告知他们的原产地。我们将采用这些方法来解决长期存在的问题 肝脏中的kupffer细胞是免疫系统中最大的髓样区室，作为病毒 面对抑制艺术的水库。我们的目标是： AIM1：在启用的主要库普弗细胞的病毒体上识别宿主细胞配体 在Ati-plasma后，具有库普弗细胞起源的病毒体的免疫亲和力分离。 AIM 2：从HIV-1感染者的肝组织中得出全长病毒信封的库 并通过高通量病毒表型和分子来评估其细胞的向流和动力学 时钟分析。 AIM 3：采用配体病毒粒子捕获和高歧视TAQ（HIDI TAQ）聚合酶定向 等位基因扩增以评估组织匹配的血浆中的病毒体是否起源于kupffer细胞和 血浆中的免疫接触病毒体和肝组织的原病毒是否表现出相关的病毒学 特征和系统发育结构。 该建议利用重要的临床资源来定义Kupffer细胞是否 用作HIV-1的病毒储藏。从该提案中得出的信息将为 识别量身定​​制的方法，以消除肝脏和其他髓样的髓样库 车厢。