Defining a Role for Liver Myeloid Cells in Viral Persistence under ART-SUPPLEMENT 1

ART-补充 1 下定义肝髓细胞在病毒持久性中的作用

• 批准号: 10852118
• 负责人: Mario Stevenson
• 金额: $ 6.02万
• 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/10852118
• 关键词: Address; Adopted; Alleles; Anatomy; Binding; CD4 Positive T Lymphocytes; Cell membrane; Cells; Cellular Tropism; Characteristics; Clinical; Disease remission; Exhibits; HIV; HIV-1; Immune system; Individual; Infection; Interruption; Kupffer Cells; Length; Libraries; Ligands; Liver; Macrophage; Membrane; Methods; Molecular; Myelogenous; Myeloid Cells; Phenotype; Phylogenetic Analysis; Plasma; Plasma Cells; Polymerase; Proviruses; Research; Resources; Role; Structure; Tissues; Viral; Viral reservoir; Viremia; Virion; Virus; antiretroviral therapy; design; molecular clock; novel; personalized approach

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感染的治疗的追求建立在支持病毒持久性的宿主的假设基础上 是可以被淘汰的。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细胞储存库的动态以及消除它们的方法。我们最近 有证据表明，在分析治疗中断(ATI)后，反弹的病毒血症由病毒粒子组成 对巨噬细胞有高度亲和性。通过使用一种新的免疫亲和浓缩方法，我们进一步 证实ATI后血浆中的嗜巨噬细胞病毒起源于髓系细胞[1]。这些 观察将髓系细胞定义为抑制ART下的储存库，这些储存库需要 关于设计病毒缓解和治疗策略的思考 ART下存放HIV-1病毒储存库的髓细胞室尚不为人所知。然而，我们的方法是 发达国家有能力提供这些信息。这些方法取决于我们的演示 病毒粒子的膜来源于宿主细胞的质膜，因此，病毒粒子含有 告知其宿主细胞的来源。我们将采用这些方法来解决长期存在的问题，即 肝脏中的库普弗细胞是免疫系统中最大的髓系细胞，起病毒的作用。 水库面对压抑的艺术。我们的目标是： 目的：鉴定来源于原代Kupffer细胞的病毒粒子上的宿主细胞配体，使其能够 ATI后血浆中来源于Kupffer细胞的病毒粒子的免疫亲和分离。 目的2：从HIV-1感染者的肝组织中提取全长病毒包膜文库 并通过高通量病毒表型和分子生物学方法评估它们的细胞亲和性和动力学 分别进行了时钟分析。 目的3：利用连接的病毒粒子捕获和高分辨Taq(Hidi Taq)聚合酶 等位基因扩增评估组织匹配血浆中的病毒粒子是否起源于库普弗细胞和 来自血浆和肝组织的免疫捕获的病毒粒子是否表现出相关的病毒学 特征和系统发育结构。 这项提议利用了重要的临床资源，这些资源将被用来定义库普弗细胞 作为HIV-1的病毒库。从这项提案中获得的信息将为 确定消除肝脏和其他髓系内髓系储存库的量身定制的方法 车厢。