Exploration into the Forgotten HIV Reservoir with Models of HIV/SIV Persistence in Mucosal Tissues

项目摘要/摘要 对于许多艾滋病毒感染者来说，终身联合抗逆转录病毒疗法(CART)已经将艾滋病毒 流行病从死刑转变为一种可控制的慢性病。然而，艾滋病毒宿主在所有人中都存在 在绝大多数患者中，接受治疗的个体和病毒反弹发生在CART中断时。给定 与终生CART治疗相关的挑战，如依从性、病毒逃逸、毒性和成本，发现 在没有CART的情况下，根除病毒和/或诱导持续病毒学控制的新方法是非常高的 艾滋病毒领域的优先事项。组织是HIV潜伏的主要部位，也是病毒反弹的重要因素 手推车中断。我们的初步数据表明，SIV感染的肥大细胞(MC)是一种粒细胞 来自髓系祖细胞的来源可能在HIV持续存在的过程中起着重要的作用，尽管还没有被研究。 纸巾。MC对免疫和神经免疫系统都有贡献，几乎完全驻留在 结缔组织和皮肤。因此，到目前为止，MC感染HIV及其在HIV中的潜在作用的研究 在分离这些细胞和识别罕见的肿瘤病灶方面，储藏库受到了后勤困难的阻碍 艾滋病毒/SIV在组织中的感染，特别是在治疗期间。我们团队的技术进步导致了 猕猴抗逆转录病毒治疗中断后反弹时SIV感染MC的鉴定。 这反过来又导致了体外研究，并由一组调查人员产生了初步数据 在艾滋病毒和MC方面的专业知识。我们的初步数据有力地支持了组织MC在HIV发病机制中的作用 和坚持不懈。这些组织驻留细胞的寿命比淋巴细胞长得多，并且在 至少在某种程度上它可以与CD4+T细胞相媲美。因此，我们建议使用各种独特的工具和 探索HIV体外感染的动力学并解决HIV感染MC的假设的资源 在HIV组织库中扮演着重要的角色。具体地说，我们将使用非人类灵长类动物的组织 研究(目标1)将通过PET-CT引导采样从确定为SIV活跃的地区收集 表情。我们将表征这些地区感染CD4+T细胞和MC的频率 不同类型的干预措施。我们将使用人体肠道和皮肤的体外和体外感染(目标2) MC了解艾滋病毒感染和持久性的动态以及不同刺激对艾滋病毒的影响 在有无购物车的情况下进行复制。最后，我们将使用MC的人性化小鼠模型(目标3)来 研究与Aim 2在体内或体外解决的问题类似的问题。总而言之，我们将 利用不同和互补的模型和技术工具箱来解决组织MC在 有助于艾滋病毒的持续存在，并确定如何操纵和瞄准这个被遗忘的水库，以 促进制定新的、更全面的艾滋病毒治疗战略。

PROJECT SUMMARY/ABSTRACT For many HIV infected people, lifelong combination antiretroviral therapy (cART) has converted the HIV epidemic from death sentence into a manageable chronic disease. However, the HIV reservoir persists in all treated individuals and viral rebound occurs upon cART interruption in the vast majority of patients. Given the challenges related to lifelong cART treatment such as adherence, viral escape, toxicity, and costs, finding novel ways to eradicate the virus and/or induce sustained virologic control in absence of cART is a very high priority in the HIV field. Tissues are major sites for HIV latency and notable contributors to viral rebound after cART interruption. Our preliminary data demonstrate that SIV infected mast cells (MC), a type of granulocyte derived from myeloid progenitors may have an important, albeit understudied role in the persistence of HIV in tissues. MC contribute to both the immune and neuroimmune systems and reside almost exclusively in connective tissues and skin. Hence, until now the study of HIV infection of MC and their potential role in HIV reservoir has been thwarted by logistical difficulties in both isolating these cells and identifying rare foci of HIV/SIV infection in tissues especially during therapy. The technological advancements of our team led to the identification of SIV infected MC at the time of rebound after antiretroviral treatment interruption in macaques. This, in turn, led to ex vivo studies and the generation of preliminary data by a team of investigators with expertise in both HIV and MC. Our preliminary data strongly support a role of tissue MC in HIV pathogenesis and persistence. These tissue resident cells live much longer than lymphocytes and susceptible to infection at least in a way that it is comparable to CD4+ T cells. Hence, we propose to use a variety of unique tools and resources to explore the dynamics of HIV infection ex vivo and address the hypothesis that HIV infected MC play an important role in the HIV tissue reservoir. Specifically, we will use tissues from non-human primate studies (Aim 1) that will be collected via PET-CT-guided sampling from areas identified with active SIV expression. We will characterize the frequency of infected CD4+ T cells and MCs in these areas during different types of interventions. We will use ex vivo and in vitro infection (Aim 2) of human gut and skin derived MC to understand the dynamics of HIV infection and persistence and the impact of different stimuli on HIV replication in presence and absence of cART. Finally, we will use a humanized mouse model of MC (Aim 3) to investigate questions similar to those addressed by Aim 2 either in vivo or ex vivo. In summary, we will leverage a toolbox of different and complementary models and techniques to address the role of tissue MC in contributing to HIV persistence and to determine how to manipulate and target this “forgotten” reservoir to facilitate the development of novel, more comprehensive HIV cure strategies.