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

描述（由申请人提供）：干扰素-?？（IFN-?）以其限制正在进行的HIV复制的能力而闻名，但随着ART的出现，对IFN-？美国抗击艾滋病毒的活动减弱了。来自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IFN-？可能对HIV储存库有活性，因为在接受IFN-? ?治疗的患者中，整合HIV DNA（但不是总HIV DNA）减少了。(聚乙二醇干扰素- ? 2 a)。这项R21提案提出了CTL作为驱动IFN-? HIV整合减少的作用。治疗。我们认为IFN-？整合的HIV DNA比非整合的HIV DNA更有效地表达HIV蛋白，这将导致整合的HIV DNA比非整合的HIV DNA更优先被清除，这是先前IFN-a试验在体内发现的结果。这些发现使我们得出了首要假设：IFN-？增强免疫清除水库。我们将使用我们的潜在模型进一步探索IFN-？通过使储存库更明显或通过增强效应功能来增加免疫清除。我们的一般方法是从ART感染患者中纯化cd4和cd8，然后用IFN-？在共培养前，然后评估水库是否减少。我们将把这些结果与已知能增强HIV表达的hdac进行比较，hdac目前正在临床试验中测试其对HIV储存库的活性。hdac对抗原呈递和HIV特异性CTL的影响尚不清楚。我们将分析IFN-a是否对CD4s或cd8产生影响，以及IFN-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IFN-？在最初的试验中，反应者和无反应者阐明了油藏减少的基本步骤。我们处于一个独特的位置，可以为这个问题做出贡献，因为我们在测量整合艾滋病毒DNA方面的专业知识，以及我们检测整合水平微小变化的能力。我们设想我们的研究将导致未来更好的试验设计和新的见解，这将是根除艾滋病毒所需要的。我们的具体目标是：目标1：确定是否

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.