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: 10527635
• 负责人: Mario Stevenson
• 金额: $ 46.95万
• 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/10527635
• 关键词: AIDS/HIV problem; Address; Adopted; Alleles; Anatomy; Antibodies; Area; 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; Immune system; In Vitro; Individual; Infection; Interruption; Kupffer Cells; Length; Libraries; Ligands; Liver; Location; 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; macrophage; 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 感染治愈方法的追求是建立在支持病毒持续存在的病毒库的假设之上的。 是可以消除的。因此，大多数关于 HIV-1 治愈和缓解的研究都集中在 确定 CD4+ T 细胞库的动态以及消除它们的方法。我们最近 提供的证据表明，分析治疗中断（ATI）后，反弹的病毒血症由病毒粒子组成 那是高度巨噬细胞热带的。通过使用新型免疫亲和富集方法，我们进一步 证明 ATI 后血浆中的嗜巨噬细胞病毒具有骨髓细胞起源 [1]。这些 观察结果将骨髓细胞定义为抑制性 ART 下的储存库，并且这些储存库需要 设计病毒缓解和治愈策略时的考虑 ART 治疗下容纳 HIV-1 病毒库的骨髓区室尚不清楚。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： 病毒粒子膜源自宿主细胞质膜，因此，病毒粒子含有配体 告知其宿主细胞的起源。我们将采用这些方法来解决长期存在的问题： 肝脏中的库普弗细胞是免疫系统最大的骨髓区室，充当病毒 面对抑制性ART。我们的目标是： 目标1：鉴定源自原代库普弗细胞的病毒颗粒上的宿主细胞配体，使 在 ATI 后血浆中对具有库普弗细胞起源的病毒体进行免疫亲和分离。 目标 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评估其细胞向性和动力学 分别进行时钟分析。 目标 3：采用配体病毒颗粒捕获和高分辨 Taq (HiDi Taq) 聚合酶导向 等位基因扩增以评估组织匹配血浆中的病毒粒子是否源自库普弗细胞和 从血浆中免疫捕获的病毒粒子和从肝组织中免疫捕获的原病毒是否表现出相关的病毒学特征 特征和系统发育结构。 该提案利用了重要的临床资源，这些资源将用于定义库普弗细胞是否 作为 HIV-1 的病毒储存库。从该提案中获得的信息将为该提案提供理由 确定消除肝脏和其他骨髓细胞内的骨髓细胞储存库的定制方法 隔间。