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

摘要 尽管对艾滋病毒进行了有效的治疗，但病毒库的存在阻碍了 一种治疗艾滋病毒感染的方法。尽管大多数储备库的研究都集中在CD4+T细胞上，但这些细胞并不是主要的 中枢神经系统中的感染细胞，而是长寿的巨噬细胞和小胶质细胞被感染。此外，许多 抗逆转录病毒治疗不会渗透到大脑。此外，药物滥用常常与艾滋病毒并存。 感染并影响中枢神经系统病毒和疾病。研究病毒在大脑中的状态，以及进行受控 滥用药物的实验不能在人类身上进行。这里我们将使用高度相关的SIV- 用有效的抗逆转录病毒治疗方案治疗感染猕猴模型的三组： 接受兴奋剂甲基苯丙胺的人、接受鸦片类吗啡的人和没有接受药物的人 滥用职权。我们的实验设计将验证大脑是一个受控的储存库的假设 治疗方案，并确定滥用药物对这种储蓄者的影响。同时，我们将 检查一个已知的储存库(淋巴中的CD4+T细胞)，作为比较，并阐明其他 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药物有效性的影响 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于 检测可能影响感染和组织迁移的细胞标志物变化的机械性研究，以及 我们对细胞内抗逆转录病毒浓度的分析在这里以及在 淋巴样细胞在药物设置中的滥用。特殊目标2将利用一系列最先进的分析方法来 检验在抑制性抗逆转录病毒治疗下，大脑确实是病毒库的假设 对于SIV和药物滥用，增加了这个储存库的大小。我们将利用聚合酶链式反应检测，病毒副产物 用指示细胞和高灵敏的Tilda试验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。为了确保具有高度敏感性和特异性的确定结果，我们将 过继将感染转移给幼稚的猴子作为新的白金标准，以确定 解决中枢神经系统作为储藏室的问题和滥用药物的影响。这些知识是至关重要的 制定治疗艾滋病毒感染的战略，包括在吸毒者中。

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.