Immune determinants of pediatric HIV/SIV reservoir establishment and maintenance

摘要-项目3 本计划项目申请的总体目标是全面了解 复杂的宿主-病原体相互作用对于HIV储存库播种和持久性至关重要， 可以制定真正针对艾滋病毒感染儿童（CLWH）独特免疫环境的战略。 我们在项目3中解决的知识差距是如何建立和维护艾滋病毒库， 由新生儿和儿童免疫系统调节，特别关注宿主和细菌代谢物， 免疫调节细胞因子和胸腺输出。艾滋病毒在成人中的持续存在是由可识别的 代谢组学和微生物谱，包括调节基因调节中的表观遗传变化的代谢物 先天/适应性免疫细胞功能和HIV潜伏期。在CLWH的研究清楚地表明， HIV库是可变，具有高动态范围。在项目3中要检验的假设是年龄- HIV储库大小和稳定性的相关变化是由影响CD 4+的特定代谢物驱动的 和CD 8 + T细胞功能，细胞因子产生（例如，IL-10和TGF-β），以及HIV感染者近期 胸腺移行细胞（RTEs）。在目标1中，我们将量化胸腺输出和感染的RTE对 完整的HIV病毒库的大小发生变化。我们将测试抗炎细胞因子的影响 在较年轻年龄组中流行的环境对胸腺输出和艾滋病毒储存库播种的影响。在目标2中，我们将 解决先天性和适应性细胞免疫稳态和HIV特异性免疫功能是如何 受宿主环境和胸腺输出的影响，使用高维流式细胞术，细胞因子 分析和转录/表观遗传分析。在目标3中，我们将解读 微生物/宿主代谢产物对IL-10和TGF-β的产生、HIV的建立和维持的影响 水库和先天/适应性免疫功能。定量短链脂肪酸的靶向代谢组学 和初级/次级胆汁酸将用于定义和验证与完整和 有翻译能力的HIV DNA。最后，免疫学、病毒学和分子数据将被整合，并且 机器学习算法将用于开发预测艾滋病毒规模和特征的模型 水库来自两个CLWH队列（IMPAACT生物储存库和EPIC 4研究）的纵向样本允许 我们监测整个儿童时期艾滋病毒储存库的动态。更好地了解艾滋病毒的脆弱性 先天性和适应性免疫压力的储存库将推动知情的方法来治愈CLWH。的 研究建议建立在我们的专业知识与尖端系统免疫学深入审问艾滋病毒。 通过多学科方法，项目和核心之间的协同作用，以及我们高度协作的 我们有信心，项目3将导致重要的发现 关于儿科HIV储库和免疫功能障碍的免疫调节。我们的使命是 将这些发现转化为临床试验方案，以推动治愈艾滋病毒感染儿童的研究。

ABSTRACT – Project 3 The overall objective of this Program project application is to generate a comprehensive understanding of the complex host-pathogen interactions critical for HIV reservoir seeding and persistence such that novel cure strategies truly targeted for the unique immune environment of children living with HIV (CLWH) can be created. The knowledge gap we address in Project 3 is how the establishment and maintenance of HIV reservoirs are regulated by the neonatal and childhood immune system, with specific focus on host and bacterial metabolites, immunoregulatory cytokines, and thymic output. HIV persistence in adults is driven by recognizable metabolomic and microbial profiles that include metabolites modulating epigenetic changes in genes regulating innate/adaptive immune cell function and HIV latency. Studies in CLWH clearly demonstrate that the size of the HIV reservoir is variable with a high dynamic range. The hypothesis to be tested in Project 3 is that age- associated changes in HIV reservoir size and stability are driven by specific metabolites that influence CD4+ and CD8+ T cell function, cytokine production (e.g., IL-10 and TGF-β), and the level of HIV-infected recent thymic emigrants (RTEs). In Aim 1, we will quantify the contribution of thymic output and infected RTEs to changes in the size of the intact HIV reservoir. We will test the impact of the anti-inflammatory cytokine environment prevalent in younger age groups on thymic output and HIV reservoir seeding. In Aim 2, we will address how innate and adaptive cellular immune homeostasis and HIV-specific immune function are influenced by the host environment and thymic output using high-dimensional flow cytometry, cytokine analyses and transcriptional/epigenetic profiling on single cells. In Aim 3, we will decipher the impact of microbial/host metabolites on production of IL-10 and TGF-β, establishment and maintenance of HIV reservoirs, and innate/adaptive immune functions. Targeted metabolomics to quantify short chain fatty acids and primary/secondary bile acids will be used to define and validate relationships with levels of intact and translation competent HIV DNA. Finally, immunological, virological, and molecular data will be integrated, and machine learning algorithms will be used to develop models to predict the magnitude and features of HIV reservoirs. Longitudinal samples from two cohorts of CLWH (IMPAACT biorepository and EPIC4 study) allow us to monitor dynamics of HIV reservoirs across childhood. A better understanding of the vulnerability of HIV reservoirs to innate and adaptive immune pressure will drive informed approaches to a cure for CLWH. The research proposed builds on our expertise with cutting-edge systems immunology to deeply interrogate HIV. With multidisciplinary approaches, synergies across Projects and Cores, and our highly collaborative group of established and early-stage investigators, we are confident that Project 3 will lead to important discoveries regarding immune regulation of the pediatric HIV reservoir and immune dysfunction. It is our mission to turn these discoveries into clinical trial protocols to advance research towards a cure for children with HIV.