Probing mechanisms of reduced HIV reservoirs in an interferon-a clinical trial

描述(申请人提供)：干扰素-？？(干扰素-？)S以其限制艾滋病毒复制的能力而闻名，但随着抗逆转录病毒药物的出现，人们对干扰素-？的抗艾滋病毒活性的兴趣减弱。最近一项试验的令人兴奋的新数据重新引起了我们的兴趣，因为它表明干扰素？在接受干扰素治疗的一部分患者中，整合的HIV DNA(而不是总的HIV DNA)减少了，因此可能对HIV宿主具有活性。(聚乙二醇化干扰素-？2a)。这项R21提案解决了CTL在推动HIV与干扰素整合减少方面的作用。心理治疗。我们认为干扰素-？可能增强免疫清除的原因是，整合的艾滋病毒DNA比非整合的艾滋病毒DNA更有效地表达艾滋病毒蛋白，这将导致整合的艾滋病毒DNA优先清除非整合的艾滋病毒DNA--这是在之前的干扰素-a试验中在体内发现的结果。这些发现使我们得出了最重要的假设：干扰素？增强宿主的免疫清除能力。我们将使用我们的潜在模型来进一步探索是否存在干扰素？通过使储存库更明显或通过增强效应器功能来增加免疫清除。我们的一般方法是从ART感染的患者中纯化CD4和CD8，并用干扰素-β单独处理这些细胞。在共培养之前，然后评估是否减少了储藏量。我们将把这些结果与HDACi进行比较，HDACi已知能增强HIV的表达，目前正在临床试验中测试其对HIV宿主的活性。HDACi对抗原提呈和HIV特异性CTL的影响尚不清楚。我们将剖析干扰素对CD4或CD8的影响，以及干扰素-a是否增强了HIV抗原的表达、呈递或CTL功能。我们还将比较和对比干扰素-？？在最初的试验中，有应答者和无应答者，以阐明减少储层的基本步骤。我们处于一个独特的地位，能够为这个问题做出贡献，因为我们在测量整合的艾滋病毒DNA方面的专业知识，以及我们检测整合水平的微小变化的能力。我们设想，我们的研究将导致未来更好的试验设计和根除艾滋病毒所需的新见解。我们的具体目标是：目标1：确定 干扰素-？通过增加HIV表达、抗原呈递和/或CTL效应来增强体外和体外潜伏细胞的清除，并与HDACi的效果进行比较，以了解干扰素-？可能会减少水库的数量。目的2：检查先前的干扰素-α试验的体内证据是否支持我们的CTL介导的清除模型。

DESCRIPTION (provided by applicant): Interferon-?? (IFN-?) is known for its ability to restrict ongoing HIV replication, but with the advent of ART, interest in IFN-?'s activity against HIV waned. Exciting new data from a recent trial renews our interest as it suggests IFN-? may have activity against HIV reservoirs since integrated HIV DNA (but not total HIV DNA) was reduced in a subset of patients treated with IFN-? (pegylated interferon-?2A). This R21 proposal addresses a role for CTL as driving the decrease in HIV integration found with IFN-? therapy. Our rationale that IFN-? may enhance immune clearance is derived from the fact that HIV proteins can be expressed more efficiently from integrated HIV DNA than from unintegrated HIV DNA, which would lead to preferential clearance of integrated over unintegrated HIV DNA-the results found in vivo in the prior IFN-a trial. These findings led us to our overarching hypothesis: that IFN-? enhances immune clearance of the reservoir. We will use our latent model to further explore if IFN-? increases immune clearance by either making the reservoir more visible or by enhancing effector function. Our general approach is to purify CD4s and CD8s from ART infected patients and to treat these cells individually with IFN-? before coculture and then assess if reservoirs are reduced. We will compare these results to HDACi which are known to enhance HIV expression and are currently being tested for activity against HIV reservoirs in clinical trials. The effects of HDACi on antigen presentation and HIV specific CTL are unknown. We will dissect if the effects are on CD4s or CD8s and if HIV antigen expression, presentation or CTL function are enhanced by IFN-a. We will also compare and contrast IFN-? Responders and Nonresponders in the original trial to elucidate the essential steps to reservoir reduction. We are in a unique position to contribute to this question because of our expertise in measuring integrated HIV DNA and our ability to detect small changes in integration levels. We envision our study will lead to future better trial design and new insights that will be required for HIV eradication. Our specific aims are: Aim 1: Determine if IFN-? enhances clearance of latent cells in vitro and ex vivo by increasing HIV expression, antigen presentation, and/or CTL effectiveness and compare to the effects of HDACi to understand how IFN-? might reduce reservoirs. Aim 2: Examine if in vivo evidence in the prior IFN-a trial supports our model of CTL- mediated clearance.