Genetically Modified Gamma Delta T Cells to Target SIV Reservoirs

目前还没有治愈艾滋病毒/艾滋病的方法。尽管抗逆转录病毒疗法(ART)对 抑制循环中的艾滋病毒水平，并不能消除潜伏和持久的病毒库和病毒 艺术中断后反弹。因此，迫切需要根治疗法来清除病毒。 并消除对终身艺术的依赖。嵌合抗原的临床研究进展 白血病和淋巴瘤中的受体(CAR)T细胞已证明重定向T细胞的CTL活性 与特定的CAR表达可以克服自体CTL功能的局限性。该项目旨在 使用γδ-CARS重定向伽马三角洲(CD4)T细胞对SHIV感染的自然细胞毒性 通过靶向HIV-Env识别受感染的细胞。CD4-CAR将与maC46表达相结合，这将 防止艾滋病毒融合，从而保护CAR-γδT细胞不受体内感染。我们在恒河猴中的数据 猕猴在淋巴结和骨髓中表现出与外周相似的γδT细胞 血液，并在肠道粘膜、肝脏、脾和肺中特别丰富，所有潜在的储存组织 抗逆转录病毒治疗期间残留的艾滋病毒感染。基于它们独特的功能特性，包括：(I)有充分的记录 肿瘤免疫治疗中的CTL活性，(Ii)组织迁移，(Iii)固有的抗HIV/SIV CTL/效应器功能， 以及(Iv)通过肠道微生物配体的持续刺激；我们假设CAR-γδT细胞将 作为有效的抗HIV CTL，抵抗HIV感染，并迁移到不依赖HIV的组织- 刺激，以持续瞄准病毒库。 在R21阶段，我们将确定自体CAR-γδT细胞过继转移的可行性 猕猴组织中的迁移和持久性及其对新城疫病毒感染的体内反应。活体内 (一)分布；(二)增殖；(三)持久性；(四)体外CTL/细胞因子效应功能 在经CD-4-CAR-γδT细胞处理的动物中进行检测，并与接受 载体转导自体γδT细胞。 在R33阶段，我们将确定针对病毒的CAR-γδT细胞免疫疗法的可行性 水库在临床背景下对感染新城疫病毒的猕猴进行有效的抗逆转录病毒治疗。领养的影响 转移的CAR-γδT细胞对病毒载量的影响将在淋巴和粘膜组织储存库中进行评估。这个 持续性和归巢、CTL活性、选择性扩增，以及最终通过 将CD4-CAR-γδT细胞与载体对照组进行比较。 由于恒河猴是HIV致病机理和基因治疗的重要临床前模型， CAR-γδT细胞免疫治疗在恒河猴/SIV模型中的评价具有较高的翻译价值。的能力 CAR-γδT细胞控制ART停用后的病毒血症将是艾滋病毒的重大进步 并将有力地推动CAR-γδT细胞免疫疗法在艾滋病毒/艾滋病中的新的临床试验。

Currently there is no cure available for HIV/AIDS. Although antiretroviral therapy (ART) is effective in suppressing circulating HIV levels, it does not eliminate the latent and persistent viral reservoirs and the virus rebounds following ART-interruption. Thus, curative therapies are urgently needed to clear the virus from infected people and eliminate the dependence on lifelong ART. Recent clinical success of Chimeric antigen receptor (CAR) T cells in leukemia and lymphoma have demonstrated that redirecting CTL activity of T cells with specific CAR expression can overcome the limitations of autologous CTL functions. This project aims to redirect the natural cytotoxicity of gamma delta (γδ) T cells against SHIV-infection using CD4-CARs that can recognize infected cells by targeting HIV-Env. The CD4-CAR will be coupled with maC46 expression that will prevent HIV fusion and thereby protect the CAR-γδ T cells from getting infected in vivo. Our data in rhesus macaques show readily available γδ T cells in lymph nodes and bone marrow at levels similar to peripheral blood, and are particularly enriched in gut mucosa, liver, spleen, and lungs, all potential reservoir tissues for residual HIV infection during ART. Based on their unique functional properties, including (i) well-documented CTL activity in immunotherapy of cancer, (ii) tissue migration, (iii) innate anti-HIV/SIV CTL/effector functions, and (iv) continued stimulation through gut microbial ligands; we hypothesize that CAR-γδ T cells will serve as potent anti-HIV CTLs resistant to HIV infection, and migrate to tissues independent of HIV- stimulation to continually target viral reservoirs. In the R21 phase, we will determine the feasibility of autologous CAR-γδ T cell adoptive transfer for migration and persistence in tissues and their in vivo response to SHIV infection in rhesus macaques. In vivo (i) distribution, (ii) proliferation, (iii) persistence; and (iv) ex vivo CTL/cytokine effector functions will be examined in the CD4-CAR-γδ T cell-treated animals and compared with the vector control group that receive vector transduced autologous γδ T cells. In the R33 phase, we will determine the feasibility of CAR-γδ T cell immunotherapy for targeting viral reservoirs in the clinical setting of SHIV-infected macaques on effective ART. The effects of the adoptively transferred CAR-γδ T cells on viral load will be evaluated in lymphoid and mucosal tissue reservoirs. The persistence and homing, CTL activity, selective expansion, and ultimately, the efficacy of adoptive transfer with CD4-CAR-γδ T cells will be compared with vector control group. Since rhesus macaque is an important pre-clinical model for both HIV pathogenesis and gene therapy, the evaluation of CAR-γδ T cell immunotherapy in rhesus/SHIV model has high translational value. The ability of CAR-γδ T cells to control viremia following withdrawal of ART would be a significant advancement in HIV treatment and would strongly promote a new clinical trial for CAR-γδ T cell immunotherapy in HIV/AIDS.