Dual CMV and HIV CARs for Cure of HIV

项目总结/摘要 CD 8+细胞毒性T淋巴细胞（CTL）在HIV-1的免疫发病机制中起着关键的保护作用 感染，但最终失败，在大多数人由于落后于病毒。许多数据表明，艾滋病毒- 1-特异性CTL介导HIV-1的抑制，HIV-1介导慢性感染的“无症状期”。 CTL应答通过幼稚细胞群的克隆扩增产生，一旦抗原被清除， 到低水平或缺乏水平，这些细胞中的大多数死亡，留下具有低效应子功能的静息中枢记忆细胞， 但其在再次攻击效应细胞时被引发以更快速地扩增。这一过程导致 CTL落后于HIV-1，病毒序列进化速度超过CTL，从而允许突变逃逸。 建立的感染可能是导致CTL耗尽的病毒持续存在的主要机制， 最终失败。虽然抗逆转录病毒疗法（ART）可以阻止大多数正在进行的病毒复制， 可能允许免疫再生，可以控制治疗中断后的感染，CTL 对静息记忆水平衰退的反应，治疗中断通常导致病毒血症迅速上升 类似于急性感染。 我们假设，持续驱动效应CTL对HIV-1的持久性可能会中断这一过程。 致病循环与其让CTL落后于HIV-1，我们提出了一种策略， 独立于HIV-1复制的HIV-1特异性效应CTL。这可以防止（在一个 未感染的人）或打破（在ART感染的人的情况下）导致CTL的致病循环 功能障碍为了实现这一目标，我们将利用人体内的慢性抗原刺激， 细胞巨细胞病毒（CMV）来驱动识别CMV和HIV-1的CTL。具体而言，我们的目标是： 1.设计慢病毒载体，产生同时靶向CMV和HIV-1的CAR T细胞; 2.确认这些载体在体外的功能，作为未来动物研究的先决条件。

PROJECT SUMMARY/ABSTRACT CD8+ cytotoxic T lymphocytes (CTLs) play a key role protective role in the immunopathogenesis of HIV-1 infection, but eventually fail in most persons due to lagging behind the virus. Numerous data indicate that HIV- 1-specific CTLs mediate the suppression of HIV-1 that mediates the “asymptomatic phase” of chronic infection. The CTL response arises by clonal expansion from the naïve cell population, and once the antigen is cleared to low or absent levels, most of these cells die, leaving resting central memory cells with low effector function, but which are primed for more rapid expansion upon re-challenge to effector cells. This process results in the lag of CTLs behind HIV-1, and viral sequence evolution outpaces CTLs to allow mutational escape during established infection that is likely a major mechanism of viral persistence leading to CTL exhaustion and eventual failure. Although treatment with antiretroviral therapy (ART) stops most ongoing viral replication and could potentially allow immune regeneration that could control infection after treatment interruption, CTL responses to decay to resting memory levels, and treatment interruption usually leads to a rapid rise in viremia similar to acute infection. We hypothesize that continuously driving persistence of effector CTLs against HIV-1 could interrupt this pathogenic cycle. Rather than allowing CTLs to lag behind HIV-1, we propose a strategy to maintain levels of HIV-1-specific effector CTLs independently of HIV-1 replication. This could prevent (in the case of an uninfected person) or break (in the case of an infected person on ART) the pathogenic cycle leading to CTL dysfunction. To achieve this goal, we will take advantage of the in vivo chronic antigenic stimulus of human cytomegalovirus (CMV) to drive CTLs that recognize both CMV and HIV-1. Specifically, we aim: 1. To engineer lentiviral vectors that generate CAR T cells targeting CMV and HIV-1 simultaneously; 2. To confirm the function of these vectors in vitro as a prerequisite for future animal studies.