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： 在没有抗逆转录病毒疗法的情况下，对艾滋病毒感染的长期自然控制的新见解将有助于推动 一种功能性治愈或“缓解”。最近的研究已经确定了HIV精英控制者(EC)的子集，他们是 被认为是“特殊的”(EEC)，具有数十年的病毒学控制而没有免疫力下降。多重 机制可能有助于控制EECS，包括强大和持续的CD8+T细胞反应。 然而，目前还不清楚EECS能否无限期地保持这种状态，他们是否有任何持续的病毒 活动并从ART的发起中受益(或因ART的撤销而受到损害)，或者它们是否自发地 清除了病毒库。这些人是否代表了持久艾滋病毒控制的真实模式，或者在某些情况下 病例中，功能性治愈是需要进一步研究的重要问题。利用最大的精英 在美国的控制员队列，加州大学旧金山分校的范围队列，我们最近的工作集中在这个问题上：是否存在 可以作为长期无抗逆转录病毒药物缓解的模型的ECs子集？使用新的完整的前病毒 DNA检测(IPDA)用于检测完整的前病毒以及敏感抗体和T细胞免疫分析，我们 确定了21个异常ECs中具有超低储集层规模的一小部分。最近的分析使用了一个组合 IPDA和Flip-Seq的研究表明，一些ECs的完整前病毒水平极低，存在于 潜伏期使它们很难被发现。因此，新的病毒学检测方法可以检测超低水平的潜伏病毒 需要使用HIV和免疫学分析来评估CD8+T细胞控制的作用，以进一步促进我们的 对EECS的理解。为了这个目标，我们建议使用我们的新的在体人化超灵敏 小鼠病毒生长试验(HmVOA)模型用于潜伏病毒检测。该模型还提供了体内独一无二的 设置以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 在有或没有被认为介导控制的自体CD8+T细胞的情况下，否则不会 可以在人类环境中进行评估。虽然对EECS的全面理解是一项复杂的任务，但在这里我们 建议从更简单的初始目标开始，以： 1.确定是否可以从杰出的精英控制者那里恢复完整的艾滋病毒(通过病毒副产物) 目前对ART和精英控制器的长期ART使用人性化的小鼠病毒生长(HmVOA) 化验 2.确定EEC中主要的病毒控制是否确实是由CD8+T细胞介导的，并评估这种控制 还可以对体内潜在的新艾滋病毒暴露(双重感染)产生抵抗力。 这些结果将有助于我们进一步了解潜伏病毒和强细胞介导的作用 EECS中的病毒控制。