Quantifying Immune VS. Viral Killing of SIV Infected and Latently Infected Cells

描述：我们建议的目的是描述CD8诱导的机会窗在恒河猴(RM)中杀死感染但尚未产生SIV的细胞的机会之窗。为此，我们将比较对照动物和CD8耗尽动物对SIV感染的治疗。尽管我们和其他人以前(部分)也做过类似的实验，但那些使用RTI疗法(PMPA和FTC)的人。在这里，我们建议使用整合酶链转移抑制剂(raltegravir-ral)，我们将获得定性和定量不同的结果，这些结果将探测细胞感染期间但在病毒产生之前(即整合前)的免疫反应强度。我们将推广标准的和成功的病毒治疗模型来分析单一治疗的病例，因为我们预计病毒的衰变行为是不同的。对CD8缺失的猕猴进行病毒载量(及其悬液)的分析，将为探讨CD8在病毒控制中的作用提供最佳条件。此外，我们将分析感染细胞的数量，以及不同形式的HIV DNA(整合的、非整合的、2ltr圈)，以获得对病毒库，特别是潜伏感染细胞的长期动态的宝贵见解。我们的具体目标是：目的1.量化SIV在恒河猴(RM)单一治疗过程中的动态。假设：RAL单一疗法下的病毒载量分布是由于感染病毒的细胞腐烂，这些细胞尚未完成整合，但会渗漏并成为生产性感染。我们将开发新的病毒动力学模型，其中考虑到RAL的特定作用模式。因此，我们将量化未整合HIV DNA的细胞储存库的动力学。目标2.量化 CD8 T细胞在SIVmac感染前(即前病毒整合前)对感染细胞的杀伤作用假设：大部分CD8T细胞具有杀伤作用 被感染的细胞在被感染的细胞开始产生病毒之前发生。我们将量化CD8对含有未整合DNA的细胞的杀伤作用。目的3.对长期SIV治疗期间不同细胞/病毒库的命运进行量化和建模。假设：长期联合治疗的模型，包括结构化的AL治疗中断，将允许描述不同细胞/病毒储存库的衰变和维持。我们将测量具有整合和非整合病毒DNA的细胞(例如，2-LTR环)、潜伏感染的细胞和滤泡树突状细胞相关病毒。我们将开发这些细胞间隔的模型来量化它们的衰变，并了解它们是在整个治疗过程中得到补充，还是持续减少。这些研究将更好地表征CD8 T细胞在控制感染和不同病毒库的长期腐烂方面的强度。因此，我们的结果将对艾滋病毒控制(通过疫苗和治疗)产生影响，并探索 根除病毒过程中的免疫反应。

DESCRIPTION: The objective of our proposal is to characterize the window of opportunity for CD8-induced killing of cells infected but not yet producing SIV in rhesus macaques (RM). To this end we will compare treatment of SIV infection in control animals and CD8-depleted animals. Although similar experiments were done before (in part) by us and others, those used RTI therapy (PMPA and FTC). Here, we propose that using an integrase strand transfer inhibitor (raltegravir - RAL), we will obtain qualitatively and quantitatively different results that will alow probing of the strength of the immune response during cell infection but before viral production (i.e., before integration). We will extend the standard and successful models of viral therapy to analyze the case of RAL monotherapy, because we expect that the behavior of the decay of virus is different. Analyzing viral load under RAL therapy (and its suspension) in CD8 depleted macaques will provide the best conditions to probe the effect of CD8s on controlling the virus. In addition, we will analyze the number of infected cells, with different forms of HIV DNA (integrated, unintegrated, 2LTR circles) to gain precious insights into the long-term dynamics of viral reservoirs, especially latently infected cells. Our specific aims are: Aim 1. Quantify the dynamics of SIV during RAL monotherapy in rhesus macaques (RM). Hypothesis: The profile of viral load under RAL monotherapy is due to decay of cells infected by virus that have not completed integration but which leak through and become productively infected. We will develop new models of viral dynamics, which take into account the specific mode of action of RAL. Thus, we will quantify the kinetics of cellular reservoirs with unintegrated HIV DNA. Aim 2. Quantify the effect of CD8+ T-cells in killing cells infected with SIVmac before they become productively infected (i.e., before proviral integration). Hypothesis: Most of the CD8+ T cell effect on killing infected cells occurs before the start of viral production by the infected cell. We will quantify te effect of CD8 on killing of cells with unintegrated DNA. Aim 3. Quantify and model the fate of different cellular/viral reservoirs during long-term SIV treatment. Hypothesis: Modeling of long-term combination therapy including structured RAL treatment interruptions will permit a characterization of the decay and maintenance of different cellular/viral reservoirs. We will measure cells with integrated and unintegrated viral DNA (eg., 2-LTR circles), latently infected cells and follicular dendritic cell-associated viruses. We will develop models of these cellular compartments to quantify their decay and understand whether they are replenished throughout treatment or if they continuously decrease. These studies will better characterize the strength of CD8+ T-cells in controlling infection and the long-term decay of different viral reservoirs. Thus, our results will have implications to HIV control (by vaccines and therapy) and probe the limits of the immune response in eradicating the virus.