Identification of SIV replication and reservoirs in the CNS

摘要 尽管高效的抗逆转录病毒疗法（ART）能够将病毒复制减少到无法检测的程度， 在感染猴免疫缺陷病毒（SIV）的猴子血浆中，我们最近开发了一种病毒包膜， 定向免疫PET/CT成像技术已经能够检测持续SIV信号的福耶斯，即使在 然而，迄今为止，这种非侵入性技术在CNS中映射SIV信号的能力 难以捉摸在本申请中，我们建议专注于探针和策略的开发， 通过优化技术，使SIV特异性PET探针能够穿过CNS， 血脑屏障和体内表达SIV env的标志细胞。为了实现这一目标，我们将使用辫子 感染SIVsmB 670/SIVmac 239 - 17 E-Fred的猕猴模型， Clements是最可靠的neuroHIV模型之一。利用实力和技术进步 在新伊比利亚研究中心和格鲁吉亚技术，我们将绘制水库播种在 中枢神经系统在急性感染和监测病毒信号的动力学后，急性感染的组合使用 免疫PET/CT，光片和共聚焦成像技术，以及感染细胞的分选，以记录 在CNS SIV感染的早期慢性阶段保持活性的细胞的性质和功能。接下来我们将 记录ART开始时以及ART后CNS中相对于全身的病毒动力学动态。 ART中断。从这些非常详细的分析中产生的数据将描绘出哪些细胞保持持久的 ART下的病毒复制（如有），更重要的是评估CNS病毒储库对 体内早期病毒反弹。对这些数据的深入分析也将为实现以下目标提供重要步骤： 开发包括CNS在内的治疗策略。

Abstract Even though highly potent antiretroviral therapy (ART) has the ability to reduce viral replication to undetectable levels in the plasma in simian immunodeficiency virus (SIV) infected monkeys, our recently developed viral env directed immunoPET/CT imaging technology has been able to detect foyers of continued SIV signals even after prolonged ART. However, the ability of this non-invasive technique to map SIV signal in the CNS have thus far been elusive. In this application we propose to focus on the development of probes and strategies to specifically address SIV signals in the CNS, by optimizing the technology to enable SIV specific PET probes to cross the blood brain barrier and mark cells expressing SIV env in vivo. Towards this goal we will use the pigtailed macaque model infected with SIVsmB670/SIVmac239-17E-Fred developed and validated by the team of Dr. Clements as one of the most reliable models of neuroHIV. Leveraging the strength and technological advances available at the New Iberia Research Center and Georgia Tech, we will map the seeding of reservoirs in the CNS during acute infection and monitor the kinetic of viral signals post-acute infection using a combination of immunoPET/CT, light-sheet and confocal imaging techniques, as well as sorting of infected cells to document the nature and function of cells remaining active in the early chronic phase of CNS SIV infection. Next we will document the dynamics of viral kinetics in the CNS relative to total body upon ART initiation, as well as post- ART interruption. Data generated from these highly detailed analyses will delineate which cells retain persistent viral replication under ART (if any) and more importantly evaluate the contribution of CNS viral reservoirs to the early viral rebound in vivo. The in depth analysis of this data will also provide important steps towards the development of cure strategies that include the CNS.