Immune determinants of pediatric HIV/SIV reservoir establishment and maintenance

摘要-项目2 本计划项目申请的总体目标是全面了解 复杂的宿主-病原体相互作用对艾滋病毒宿主的播种和持久性至关重要，因此新的治愈方法 可以创建真正针对艾滋病毒携带者儿童(CLWH)独特免疫环境的战略。 我们在项目2中解决的知识差距是如何建立和维护艾滋病毒储存库 受新生儿和儿童免疫系统的调节，特别关注溶细胞性和非溶细胞性 CD8+T细胞的抗病毒作用。CLWH储集层持久性的驱动因素还不完全清楚 儿科先天免疫系统和适应性免疫系统在不同发育阶段的复杂性 找到治疗艾滋病毒的方法的挑战。在成人模型中越来越多的证据表明CD8+T细胞是 在抗逆转录病毒疗法(ART)下维持艾滋病毒抑制所需的，我们最近发现 感染后播种的病毒库不依赖于抗原特异性的经典细胞溶解功能 CD8+T细胞。项目2中要检验的假设是CD8+T细胞介导的HIV/SIV前潜伏期 这种作用在儿科模型中是可重现的；然而，CD8+T细胞这种非经典作用的明显特征 细胞在生命早期可能受到调节性免疫环境的影响。在目标1中，我们将发展儿科 CD8+T细胞介导的HIV/SIV调控机制的体外模型研究 婴儿和儿童的潜伏期通过比较免疫表型、转录和病毒学分析 CD8+T细胞的存在或缺失。在目标2中，我们将进行体内原则验证研究，使用 实验性抗体介导的CD8+细胞在恒河猴(RM)婴儿感染前的耗竭 SIVmac239M。为了评估CD8+T细胞在SIV储备库建立中的作用，我们将比较病毒动力学 以及ART上的病毒库大小/多样性/整合位置以及ART中断后病毒的反弹 CD8耗竭和未耗竭的红斑狼疮婴儿。在目标3中，将进行抗体介导的CD8+细胞耗尽 在使用和不使用LRA的长期ART治疗后，感染SIVmac239M的RM婴儿 CD8+T细胞的清除扰乱了水库的维护。CD8+T细胞介导的潜伏期机制 逆转将通过多组学分析进一步评估。更好地了解艾滋病毒的脆弱性 先天免疫压力和适应性免疫压力将推动治疗慢性肝炎的知情方法。这个 建议的研究建立在我们在婴儿RM模型、最先进的水库分析和T细胞方面的专业知识的基础上 免疫学对儿童HIV/SIV的深入询问。借助多学科方法，实现跨领域协同 项目和核心，以及我们由成熟和早期调查人员组成的高度协作的团队，我们是 相信项目2将导致有关儿科艾滋病毒免疫调节的重要发现 蓄水池和免疫功能障碍。我们的使命是将这些发现转化为临床试验方案，以 推进治疗艾滋病毒儿童的研究。

ABSTRACT – Project 2 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 2 is how the establishment and maintenance of HIV reservoirs are regulated by the neonatal and childhood immune system, with specific focus on the cytolytic and non-cytolytic antiviral role of CD8+ T cells. The drivers of reservoir persistence in CLWH are incompletely understood and the complexity of the pediatric innate and adaptive immune system across developmental stages contributes to the challenge of a finding cure for HIV. Mounting evidence in adult models implicates CD8+ T cells as being required for maintaining HIV suppression under antiretroviral therapy (ART) and we have recently found that the viral reservoir seeded after infection does not depend on the classical cytolytic function of antigen-specific CD8+ T cells. The hypothesis to be tested in Project 2 is that the CD8+ T cell-mediated HIV/SIV pro-latency effect will be reproducible in pediatric models; however, distinct features of this non-classical role for CD8+ T cells may be influenced by the regulatory immune environment in early life. In Aim 1, we will develop pediatric in vitro models to thoroughly investigate the mechanisms involved in CD8+ T cell-mediated control of HIV/SIV latency in infants and children via comparative immunophenotyping, transcriptomic, and virological analyses in presence or absence of CD8+ T cells. In Aim 2, we will conduct a proof-of-principle in vivo study using experimental antibody mediated CD8+ cell depletion in rhesus macaque (RM) infants prior to infection with SIVmac239M. To assess the role of CD8+ T cells in SIV reservoir establishment we will compare viral dynamics and viral reservoir size/diversity/integration sites on ART and virus rebound after ART interruption between CD8-depleted and undepleted RM infants. In Aim 3, antibody-mediated CD8+ cell depletion will be performed in SIVmac239M-infected RM infants after long-term ART with and without an LRA to test the extent to which removal of CD8+ T cells disrupts reservoir maintenance. CD8+ T cell-mediated mechanisms involved in latency reversal will be further assessed through multiomic analyses. 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 infant RM models, state-of-the-art reservoir assays, and T cell immunology to deeply interrogate pediatric HIV/SIV. With multidisciplinary approaches, synergies across Projects and Cores, and our highly collaborative group of established and early-stage investigators, we are confident that Project 2 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.