The brain as a SIV reservoir under suppressive cART potentiation by drugs of abuse

摘要 尽管对艾滋病毒有有效的治疗，但病毒储库的存在阻止了艾滋病的发展。 艾滋病病毒感染的治疗方法虽然大多数储库工作都集中在CD 4 + T细胞，但这些不是主要的储库。 CNS中的受感染细胞而不是长寿的巨噬细胞和小胶质细胞被感染。此外，许多 抗逆转录病毒疗法不会穿透大脑。此外，药物滥用经常与艾滋病毒并发 感染并影响CNS病毒和疾病。研究病毒在大脑中的状态， 滥用药物的实验，不能在人类身上进行。在这里，我们将使用高度相关的SIV- 感染的猕猴模型用三组有效的抗逆转录病毒治疗方案治疗： 接受兴奋剂甲基苯丙胺，那些接受阿片类吗啡和那些没有接受药物 虐待我们的实验设计将检验大脑是一个受控制的储存库的假设 治疗方案，并确定滥用药物对这种储库的影响。同时，我们将 检查一个已知的水库（淋巴结CD 4 + T细胞），既作为比较，并阐明额外的 滥用药物对水库的影响。 具体目标1将审查滥用药物对调节 SIV感染猕猴的抗逆转录病毒治疗。创新方面包括我们的假设基础 检查可能影响感染和组织迁移的细胞标志物变化的机制研究，以及 我们对细胞内抗逆转录病毒浓度的分析在脑细胞中进行， 淋巴细胞在滥用药物的情况下。Specific Aim 2将利用一系列最先进的检测方法， 检验在抑制性抗逆转录病毒治疗下，大脑确实是病毒储存库的假设 SIV和药物滥用增加了这个水库的大小。我们将利用PCR检测，病毒生长 在指示细胞和高灵敏度TILDA试验中检测纯化的小胶质细胞/巨噬细胞中的病毒 从目标1中的动物的大脑中提取。为了确保高灵敏度和特异性的明确结果，我们将 将感染过继性转移给未感染的猴子作为新的铂标准用于水库检测， 解决中枢神经系统作为贮存器和滥用药物的影响的问题。这些知识至关重要 制定治疗艾滋病毒感染的战略，包括吸毒者。

Abstract Despite effective treatments for HIV, the presence of viral reservoirs has prevented the development of a cure for HIV infection. Although most reservoir work has focused on CD4+ T cells, these are not the primary infected cells in the CNS instead long-lived macrophages and microglia are infected. Furthermore, many of the anti-retroviral treatments do not penetrate the brain. In addition, drug abuse is frequently co-morbid with HIV infection and affects CNS virus and disease. Studying the state of virus in the brain, as well as doing controlled experiments with drugs of abuse, cannot be performed in humans. Here we will use the highly relevant SIV- infected macaque model treated with an effective anti-retroviral treatment regimen in three groups: those receiving the stimulant methamphetamine, those receiving the opiate morphine and those receiving no drugs of abuse. Our experimental design will test the hypothesis that the brain is a reservoir under controlled treatment regimens and determine the effects drugs of abuse have on such a reservoir. In parallel, we will examine a known reservoir (lymph node CD4+ T cells), both as a comparison and to illuminate additional effects of drugs of abuse on reservoirs. Specific Aim 1 will examine the effects of drugs of abuse on modulating the effectiveness of antiretroviral treatment in SIV infected macaques. Innovative aspects include our hypothesis based mechanistic studies examining changes in cellular markers that may impact infection and tissue migration, and our analyses of intracellular antiretroviral concentrations uniquely performed here in brain cells as well as in lymphoid cells in the setting of drugs of abuse. Specific Aim 2 will utilize a series of state of the art assays to examine the hypotheses that under suppressive anti-retroviral treatment, the brain is indeed a viral reservoir for SIV and substance abuse increases the size of this reservoir. We will utilize PCR detection, viral outgrowth in indicator cells and the highly sensitive TILDA assays to examine virus in microglia/macrophages purified from the brains of the animals in Aim 1. To ensure a definite result with high sensitivity and specificity, we will adoptively transfer infection to naïve monkeys as a new platinum standard for reservoir detection to definitely address this question of the CNS as a reservoir and the effects of drugs of abuse. Such knowledge is crucial for devising strategies to cure HIV infection, including in drug abusers.