Induction/Eradication of SIV in an Optimized HAART CNS Macaque Model

虽然高效抗逆转录病毒疗法对艾滋病流行病产生了重大影响，但它不会终结这一流行病；需要的是清除体内的感染，这就需要使用药物进入骨髓，大脑，任何潜伏艾滋病毒的组织，清除或降低这些病毒库，使HAART成为不必要的。设计抗逆转录病毒药物不需要动物模型，但动物模型是测试药物诱导潜伏HIV和根除潜伏病毒库能力的重要临床前步骤。由于这些药物的潜在毒性和未知的疗效，有效的动物模型对于临床试验设计中诱导/根除潜伏性HIV的治疗药物的测试至关重要。我们在hiv感染者中建立了严格的HAART SIV模型，将外周血和脑脊液（CSF）中的病毒载量降低到无法检测的水平，并且在组织、脑和脾脏中的CD4+ T细胞和巨噬细胞中存在潜伏病毒。由于大脑是病毒潜伏的部位，根除艾滋病毒需要HAART，其中包括cns渗透的抗逆转录病毒药物（ARTs）。我们将在SIV猕猴身上使用现有的抗逆转录病毒药物来优化一种cns渗透的HAART方案，确认药物在组织中达到有效水平，定量测定每种抗逆转录病毒药物的CSF和脑水平。使用SIV HAART模型，假设单核细胞/巨噬细胞谱系和CD34+细胞中存在潜在的病毒库（除了CD4+ T细胞），并且需要动物模型来识别，定量和表征这些库。我们假设，病毒感染可以通过诱导潜伏感染细胞的病毒再激活，并通过有效渗透组织（包括大脑）的抗逆转录病毒药物阻止病毒生命周期的完成，从组织中根除。我们将在这个SIV模型中描述CD4+ T细胞、单核细胞、组织巨噬细胞和OD34+细胞（脑、脾、骨髓、淋巴结、肺和肠道）中的潜伏库。一旦潜伏感染的静息CD4+ T细胞数量在血浆中达到稳定水平，我们将在感染siv的恒河猴中使用HAART测试诱导/根除药物，定量检测诱导治疗前后血浆中残留的病毒复制和潜伏感染的细胞，CSF和脾脏活检。我们将检查HAART停药后病毒再激活的时间，成功的诱导/根除治疗可能不会导致病毒再激活。我们将使用这样一个成功的治疗方案来剖析这种诱导/根除方法的分子基础。

While HAART has significantly impacted the AIDS epidemic it will not end the epidemic; the need is to purge the body of infection which will require the use of drugs that reach into bone marrow, brain, any tissue that harbors latent HIV and clear or lower these viral reservoirs to make HAART unnecessary. Animal models were not required for design of antiretroviral drugs, but are an essential preclinical step to test the ability of drugs to induce latent HIV and eradicate latent viral reservoirs. Because of potential toxicity and unknown efficacy of such drugs, an effective animal model is critical for testing of therapeutic agents that will induce/eradicate latent HIV for design of clinical trials. We have a rigorous SIV model of HAART in HIVinfected individuals that reduces viral load in peripheral blood and cerebrospinal fluid (CSF) to undetectable levels and have latent virus in CD4+ T cells and macrophages in tissues, brain and spleen. HIV eradication requires HAART that includes CNS-penetrant antiretrovirals (ARTs) since brain is a site of viral latency. We will optimize a CNS-penetrant HAART regimen using current ART drugs using our SIV macaque, confirming that drugs reach effective levels in tissues quantitating CSF and brain levels of each antiretroviral drug. Using this SIV HAART model, the hypothesis that latent viral reservoirs (in addition to CD4+ T cells) in cells ofthe monocyte/macrophage lineage and potentially in CD34+ cells exist, and that an animal model is required to identify, quantitate and characterize these reservoirs. We hypothesize that viral infection can be eradicated from tissues by inducing virus reactivation from latently infected cells and preventing completion ofthe virus life cycle by ART that effectively penetrate tissues, including brain. We will characterize latent reservoirs in this SIV model in CD4+ T cells, monocytes, tissue macrophages and OD34+ cells (brain, spleen, bone marrow, lymph nodes, lung and gut). We will test induction/eradication drugs in SIV-infected macaques with HAART once the number of latently infected resting CD4+ T cells plateaus in plasma, quantitate residual virus replication and latently infected cells in plasma, CSF and spleen biopsies before and after induction therapy. We will examine the time to virus reactivation after withdrawal from HAART and successful induction/eradication therapy may result in no virus reactivation. We will use such a successful treatment regimen to dissect the molecular basis ofthis induction/eradication approach.