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.

对HIV-1感染治愈的追求是建立在这样一种假设之上的，即支持病毒持续存在的宿主 是可以被消灭的因此，大多数关于HIV-1治愈和缓解的研究都集中在 确定CD 4 + T细胞库的动态以及消除它们的方法。我们最近 提出的证据表明，分析治疗中断（ATI）后，反弹病毒血症包括病毒粒子 它们是高度嗜巨噬细胞的。通过使用一种新的免疫亲和富集方法，我们进一步 证明ATI后血浆中的嗜巨噬细胞病毒具有髓样细胞来源[1]。这些 观察将髓系细胞定义为抑制性ART下的储库，并且这些储库需要 设计病毒缓解和治愈策略时的考虑 在ART下容纳HIV-1储库的髓样区室是未知的。然而，我们的方法 发达国家有能力提供这些信息。这些方法取决于我们的证明， 病毒体膜来源于宿主细胞质膜，因此，病毒体携带配体， 告知其宿主细胞来源。我们会采用这些方法，以解决长期存在的问题， 肝脏中的库普弗细胞是免疫系统中最大的髓样区室， 我们的目标是： 目的1：鉴定来源于原代库普弗细胞的病毒体上的宿主细胞配体， ATI后血浆中库普弗细胞来源的病毒粒子的免疫亲和分离。 目的2：从HIV-1感染者的肝组织中获得全长病毒包膜文库 并通过高通量病毒表型分析和分子生物学方法评估其细胞向性和动力学。 时钟分析。 目的3：采用配体病毒粒子捕获和高分辨Taq（HiDi Taq）聚合酶定向 等位基因扩增以评估组织匹配血浆中的病毒体是否来源于枯否细胞， 来自血浆的免疫捕获的病毒粒子和来自肝组织的前病毒是否表现出相关的病毒学特性， 特征和系统发育结构。 这项提案利用了重要的临床资源，将用于确定库普弗细胞是否 作为HIV-1的病毒库。从本提案中获得的信息将为 确定消除肝脏和其他骨髓内骨髓储库的定制方法 隔间