Exploring the features of HIV exceptional elite controllers in humanized mice

探索人源化小鼠中艾滋病毒杰出精英控制者的特征

• 批准号: 10472752
• 负责人: Ramesh Akkina
• 金额: $ 19.34万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10472752
• 关键词: Animal Model; Antibodies; Area; Autologous; Biological Assay; CD34 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Clinical; Colorado; Complex; Comprehension; DNA; Detection; Disease remission; Dose; Enterochromaffin Cells; Evaluation; Exhibits; Genetic Transcription; Goals; HIV; HIV Infections; HIV drug resistance; Hematopoietic stem cells; Human; Immune; Immune response; Immunologics; Immunology procedure; In Vitro; Individual; Infection; Inflammation; Investigation; Mediating; Modeling; Mus; Peripheral Blood Mononuclear Cell; Population; Proviruses; Recommendation; Residual state; Resistance; Resources; Role; Site; T cell response; T-Lymphocyte; Testing; Time; Umbilical Cord Blood; Universities; Viral; Viral Physiology; Viral reservoir; Virus; Virus Latency; Withdrawal; Work; base; cohort; experience; humanized mouse; in vivo; insight; novel; reconstitution; superinfection; viral detection

Project Summary/Abstract: Newer insights into long-term natural control of HIV infection in the absence of ART will inform efforts to induce a functional cure or “remission.” Recent work has identified a subset of HIV elite controllers (EC) who are considered “exceptional” (EEC) with decades of virologic control without immunologic decline. Multiple mechanisms likely contribute to control in EECs, including a potent and sustained CD8+ T cell response. However, it is unclear whether EECs can maintain this status indefinitely, whether they have any ongoing viral activity and benefit from initiation of ART (or are harmed by withdrawal of ART), or whether they spontaneously cleared viral reservoirs. Whether such individuals may represent a true model of durable HIV control or, in some cases, a functional cure are important questions needing further investigation. Leveraging the largest elite controller cohort in the USA, the UCSF SCOPE cohort, our recent work focused on the question: is there a subset of ECs that might serve as a model for long-term ART-free HIV remission? Using the new intact proviral DNA assay (IPDA) to detect intact proviruses as well as sensitive antibody and T-cell immune-assays, we identified a small subset of 21 exceptional ECs with ultra-low reservoir size. Recent analyses using a combination of IPDA and FLIP-seq suggested that some ECs have ultralow levels of intact proviruses existing in a deeply latent state making their detection difficult. Therefore, novel virological assays to detect ultralow levels of latent HIV and immunological analyses to evaluate the role of CD8+ T cell control need to be employed to further our understanding of EECs. Towards this goal, we propose to employ our new ultrasensitive in vivo humanized mouse viral outgrowth assay (hmVOA) model for latent viral detection. This model also provides a unique in vivo setting to examine the role of immune control since CD4+ T cells from the EEC can be tested for viral outgrowth in the presence or absence of autologous CD8+ T cells believed to mediate control which is otherwise not possible to evaluate in a human setting. While full comprehension of EECs is a complex undertaking, here we propose to start with simpler initial goals to: 1. Determine whether intact HIV can be recovered (via viral outgrowth) from exceptional elite controllers not currently on ART and elite controllers on long-term ART using the humanized mouse viral outgrowth (hmVOA) assay 2. Ascertain if the predominant viral control in EEC is indeed mediated by CD8+ T cells and assess if this control can also provide resistance to a potential new HIV exposure (superinfection) in vivo. Results from here will help in furthering our understanding of the latent virus and the role of strong cell-mediated virus control in EECs.

项目摘要/摘要： 在缺乏抗逆转录病毒治疗的情况下对艾滋病毒感染进行长期自然控制的新见解将为诱导努力提供信息 功能性治愈或“缓解”。最近的工作已经确定了 HIV 精英控制者 (EC) 的一个子集，他们是 被认为是“例外”（EEC），数十年的病毒学控制而没有免疫学下降。多种的 机制可能有助于控制 EEC，包括有效且持续的 CD8+ T 细胞反应。 然而，目前尚不清楚 EEC 是否可以无限期地维持这种状态，以及它们是否有任何持续的病毒感染。 开始 ART 的活动和益处（或因停止 ART 受到损害），或者是否自发地 清除病毒库。这些人是否可以代表持久艾滋病毒控制的真正典范，或者在某些方面 在这些病例中，功能性治愈是需要进一步研究的重要问题。撬动最大规模的精英 美国的控制者队列，UCSF SCOPE 队列，我们​​最近的工作重点是这个问题：是否存在一个 EC 的子集可以作为长期无 ART 艾滋病毒缓解的模型吗？使用新的完整原病毒 DNA 测定 (IPDA) 用于检测完整的原病毒以及敏感的抗体和 T 细胞免疫测定，我们 确定了 21 个具有超小储层尺寸的特殊 EC 的一小部分。最近使用组合进行的分析 IPDA 和 FLIP-seq 的结果表明，一些 EC 存在超低水平的完整原病毒，存在于深层 潜伏状态使得它们的检测变得困难。因此，新的病毒学检测方法可以检测超低水平的潜伏病毒 需要采用 HIV 和免疫学分析来评估 CD8+ T 细胞控制的作用，以进一步推进我们的研究。 对 EEC 的了解。为了实现这一目标，我们建议采用我们新的超灵敏体内人源化 用于潜伏病毒检测的小鼠病毒生长测定（hmVOA）模型。该模型还提供了独特的体内 设置检查免疫控制的作用，因为可以测试来自 EEC 的 CD4+ T 细胞的病毒生长情况 在存在或不存在自体 CD8+ T 细胞的情况下，据信可以介导控制，否则则不会 可以在人类环境中进行评估。虽然全面理解 EEC 是一项复杂的任务，但在这里我们 建议从更简单的初始目标开始： 1. 确定完整的艾滋病毒是否可以从特殊的精英控制者那里恢复（通过病毒生长） 目前正在接受 ART 治疗，精英控制者则使用人源化小鼠病毒生长物 (hmVOA) 进行长期 ART 治疗 化验 2. 确定 EEC 中的主要病毒控制是否确实由 CD8+ T 细胞介导，并评估这种控制是否有效 还可以对体内潜在的新的 HIV 暴露（重复感染）提供抵抗力。 这里的结果将有助于进一步了解潜伏病毒和强细胞介导的作用 东部经济走廊中的病毒控制。