The Role of Follicular Dendritic Cells in Maintenance of Persistent HIV

尽管抗逆转录病毒疗法（ART）有效地控制了HIV-1感染，但它有一些局限性 包括不舒服的副作用、低可及性和高成本。的发展 消毒或功能性固化将解决这些限制中的许多。主要障碍是 治疗是在ART患者中存在持续的、有复制能力的HIV。最近， 临床试验表明，他们可以用小分子抑制剂靶向持续存在的艾滋病病毒。 然而，这项工作的大部分集中在病毒，这是很容易获得的， 循环系统下一个重要步骤是具体确定 在接受抗逆转录病毒治疗的患者中存在持续的艾滋病毒，因此新的治疗方法可以针对这些部位。 该项目的重点是淋巴结滤泡，这是一个已知的持久性HIV-1的来源。 然而，淋巴结滤泡中的单个细胞类型在HIV持续存在中的作用是 没有完全定义。淋巴结滤泡含有滤泡树突状细胞（FDC）， 在它们的细胞表面隔离有复制能力的病毒，然后这些病毒可能感染 易感滤泡性T辅助细胞（TFH）。这取决于淋巴中的艾滋病病毒 淋巴结主要由靶细胞中潜伏的HIV前病毒接种，或者是否与FDC相关 病毒体对持久性的贡献极大地影响了未来旨在清除 这种病毒应该被开发出来。为了解决这个问题，我们收集了淋巴结滤泡 从长期ART的研究参与者，并将表征FDC的形态， TFH使用显微镜。然后，我们将使用激光捕获分离FDC和TFH 显微切割，并将使用单基因组测序和单前病毒测序， 表征这些细胞中的病毒，并确定隔离在FDC上的HIV是否作为 接受抗逆转录病毒疗法的病人体内的传染性病毒储存库，以及这种病毒是否会传播到 其他已知的持久性艾滋病病毒库。通过定义FDC在持久性和 艾滋病毒在长期接受抗逆转录病毒治疗的患者中传播，这项重要的工作将有助于 发展未来的艾滋病毒根除临床试验，并使他们能够集中和指导 治疗的解剖位置，是最相关的持久性艾滋病毒的来源。 此外，该项目将专门针对NIAID的重点领域“治愈艾滋病毒 感染”的优先事项之一，以确定是否存在其他艾滋病毒感染的水库。

Although anti-retroviral therapy (ART) effectively controls HIV-1 infection, it has several limitations including uncomfortable side effects, low accessibility, and high cost. The development of a sterilizing or functional cure would address many of these limitations. The primary obstruction to a cure is the presence of persistent, replication-competent HIV in patients on ART. Recently, several clinical trials have shown that they can target persistent HIV with small-molecule inhibitors. However, most of this work has focused on virus that is pharmacologically easily accessible in the circulatory system. The next important step is to specifically determine where reservoirs of persistent HIV exist in patients on ART so that new therapies can be targeted to these locations. This project focuses on the lymph node follicles, which are a known source of persistent HIV-1. However, the role of the individual cell types in the lymph node follicles in the persistence of HIV is not fully defined. The lymph node follicles contain follicular-dendritic cells (FDCs) that can sequester replication competent virus on their cell surface, which could then potentially infect susceptible follicular T helper cells (TFHs). Depending on whether the persistent HIV in the lymph nodes is primarily seeded by latent HIV proviruses in target cells or whether FDC-associated virions are contributing to persistence drastically affects how future strategies aimed at clearing this virus should be developed. To address this question, we have collected lymph node follicles from study participants on long-term ART and will characterize the morphology of the FDCs and TFHs using microscopy. Then, we will isolate both the FDC and the TFHs using laser-capture microdissection and will use single-genome sequencing and single-proviral sequencing to characterize the virus in these cells and determine whether HIV sequestered on FDCs acts as a reservoir of infectious virus in patients on ART and whether this virus can then disseminate to other known reservoirs of persistent HIV. By defining the role of FDCs in the persistence and dissemination of HIV in patients on long-term ART, this important work will contribute to the development of future HIV eradication clinical trials and allow them to focus and direct therapeutics to the anatomical locations that are the most relevant sources of persistent HIV. Additionally, this project will specifically address the NIAIDs area of emphasis to “Cure HIV Infection” priority one to determine if additional reservoirs of HIV infection exist.