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

量化免疫VS。

• 批准号: 8603535
• 负责人: Ruy M Ribeiro
• 金额: $ 64.45万
• 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 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603535
• 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：我们提案的目的是表征cd8诱导杀死恒河猴（RM）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缺失动物的SIV感染治疗。虽然我们和其他人之前（部分）做过类似的实验，但那些使用RTI疗法（PMPA和FTC）。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，这将允许在细胞感染期间但在病毒产生之前（即在整合之前）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单药治疗的情况，因为我们预计病毒的衰变行为是不同的。在CD8缺失的猕猴体内分析RAL治疗（及其悬浊液）下的病毒载量将为探索CD8控制病毒的作用提供最佳条件。此外，我们将分析具有不同形式HIV DNA（整合，非整合，2LTR环）的感染细胞的数量，以获得对病毒储存库，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：目标1。量化恒河猴（RM）在RAL单药治疗期间SIV的动态。假设：在RAL单一疗法下，病毒载量的概况是由于被病毒感染的细胞的衰变，这些细胞尚未完成整合，但泄漏出来并成为有效感染。我们将开发新的病毒动力学模型，其中考虑到RAL的具体作用模式。因此，我们将量化带有未整合HIV DNA的细胞储存库的动力学。目标2。量化