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

量化免疫VS。

• 批准号: 8885643
• 负责人: Ruy M Ribeiro
• 金额: $ 69.47万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8885643
• 关键词: AIDS/HIV problem; Accounting; Animals; Behavior; Biology; CD8B1 gene; Cell model; Cells; Combined Modality Therapy; Communicable Diseases; Control Animal; Coupled; DNA; DNA Integration; Data; Ethics; Follicular Dendritic Cells; HIV; Human; Immune; Immune response; Immune system; Infection; Infection Control; Integrase; Integrase Inhibitors; Interruption; Kinetics; Lead; Macaca; Macaca mulatta; Maintenance; Measures; Modeling; Morbidity - disease rate; Pathogenesis; Population; Process; Production; Protease Inhibitor; Protocols documentation; RNA-Directed DNA Polymerase; SIV; Satellite Viruses; Structure; Suspension substance; Suspensions; T-Lymphocyte; Therapeutic Agents; Time; Vaccine Therapy; Viral; Viral Load result; Viral Proteins; Virus; cell killing; in vivo; inhibitor/antagonist; insight; interdisciplinary approach; killings; mathematical model; mortality; pinacolyl methylphosphonic acid; prevent; public health relevance; research study; viral DNA

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.

描述：我们提案的目的是描述 CD8 诱导杀死恒河猴 (RM) 中已感染但尚未产生 SIV 的细胞的机会窗口。为此，我们将比较对照动物和 CD8 耗尽的动物对 SIV 感染的治疗。尽管我们和其他人之前（部分）做过类似的实验，但这些实验使用的是 RTI 疗法（PMPA 和 FTC）。在这里，我们建议使用整合酶链转移抑制剂（拉替拉韦 - RAL），我们将获得定性和定量不同的结果，这将允许在细胞感染期间但在病毒产生之前（即整合之前）探测免疫反应的强度。我们将扩展病毒治疗的标准和成功模型来分析 RAL 单一疗法的情况，因为我们预计病毒的衰变行为是不同的。分析 CD8 耗尽的猕猴在 RAL 治疗（及其悬浮液）下的病毒载量将为探讨 CD8 控制病毒的效果提供最佳条件。此外，我们将分析具有不同形式的 HIV DNA（整合的、未整合的、2LTR 环）的感染细胞的数量，以获得对病毒库的长期动态的宝贵见解，尤其是潜伏感染的细胞。我们的具体目标是： 目标 1. 量化恒河猴 (RM) RAL 单药治疗期间 SIV 的动态。假设：RAL 单一疗法下的病毒载量分布是由于病毒感染的细胞腐烂而导致的，这些病毒尚未完成整合，但渗漏并被有效感染。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，我们将量化未整合的 HIV DNA 细胞库的动力学。目标 2. 量化 CD8+ T 细胞在有效感染 SIVmac 之前（即前病毒整合之前）杀死感染 SIVmac 的细胞的作用。假设：大部分CD8+T细胞起杀伤作用 受感染的细胞发生在受感染的细胞开始产生病毒之前。我们将量化 CD8 对杀死具有未整合 DNA 的细胞的作用。目标 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（包括结构化 RAL 治疗中断）的建模将允许表征不同细胞/病毒库的衰减和维持。我们将测量具有整合和未整合病毒DNA（例如2-LTR环）的细胞、潜伏感染的细胞和滤泡树突细胞相关病毒。我们将开发这些细胞区室的模型，以量化它们的衰减并了解它们是否在整个治疗过程中得到补充或是否持续减少。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衰减方面的强度。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，并探讨艾滋病毒的局限性 消灭病毒的免疫反应。