Targeted cell-depleting immunotherapy for TB and HIV

结核病和艾滋病毒的靶向细胞消耗免疫疗法

• 批准号: 10012368
• 负责人: WILLIAM Ramses BISHAI
• 金额: $ 67.28万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-25 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10012368
• 关键词: Acute; Address; Animals; Bacille Calmette-Guerin vaccination; Biological Products; CD4 Positive T Lymphocytes; Cells; Chimeric Proteins; Chronic; Containment; Data; Denileukin Diftitox; Diphtheria Toxin; Disease; FOXP3 gene; Granuloma; HIV; HIV Infections; Human; IL2 gene; IL2RA gene; IL4 gene; IL4R gene; ITGAM gene; Immunosuppressive Agents; Immunotherapy; Indoles; Lead; Lung; Maintenance; Methods; Modeling; Mus; Mycobacterium tuberculosis; Myeloid-derived suppressor cells; Necrosis; Oryctolagus cuniculus; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacotherapy; Play; Process; Proliferating; Regulatory T-Lymphocyte; Research; Role; Spleen; Suppressor-Effector T-Lymphocytes; System; Technology; Tuberculosis; Vaccine Adjuvant; Virus Replication; cancer therapy; global health; improved; novel; novel therapeutics; pathogen; prevent; reactivation from latency; single-cell RNA sequencing; tuberculosis drugs; tuberculosis granuloma; tuberculosis immunity

PROJECT SUMMARY Tregs and MDSCs have emerged as key suppressor cells that inhibit effector immunity in the TB granuloma. In addition, Tregs have been implicated in the maintenance of HIV latency. We have generated two selective biologic agents for potent depletion of Tregs and and MDSCs using diphtheria toxin (DT) fusion protein technology: DT-IL2 targets IL2-R-positive (CD25+) cells (Tregs), and DT-IL4 targets IL4-R positive (CD124+) cells (MDSCs). Our preliminary data demonstrate that: (i) DT-IL2 and DT-IL4 reduce the abundance of Treg and MDSC cells, respectively, in mouse lungs and spleens, (ii) DT-IL2 and DT-IL4 reduces the ability of M. tb to proliferate in an acute murine TB infection model, (iii) the drug, tasquinimod which has potent anti-MDSC activity, reduced the ability of M. tb to proliferate in an acute TB infection model with a concomitant reduction in MDSCs, and (iv) administration of DT-IL2 to CD4+ T cells from patients with controlled HIV reactivates viral replication. These data support the hypotheses that (1) Tregs and MDSCs play important inhibitory roles in M. tb containment, and (2) that Treg cells play a role in HIV latency maintenance. A central scientific premise of this application is that further definition of the immunosuppressive roles of Tregs and MDSCs in TB and HIV infection will reveal important immunopathogenesis mechanisms that may be exploited towards developing improved host-directed therapies (HDTs) for both diseases. In this proposal we seek to address this premise by pursuing the following aims. Aim 1: define the role of Treg cells in the TB granuloma using targeted CD25+ cell depletion with DT-IL2. We will (a) evaluate the cellular composition of both cellular (C57BL/6, WT which are sst1R) and necrotic (C57BL/6 sst1S) murine granulomas with and without CD25+ cell depletion using single-cell RNA-Seq (SSRS) as well as traditional methods, (b) evaluate the efficacy of targeted CD25+ cell depletion in chronic murine TB with and without anti-TB drug therapy, and (c) evaluate the contribution of Treg cells to the process of cavitation (granuloma breakdown) using a novel model of rabbit cavitary TB. Aim 2: define the role of MDSCs in the TB granuloma using targeted CD124+ cell depletion with DT-IL4. We will (a) evaluate the cellular composition of both cellular and necrotic murine granulomas with and without CD124+ cell depletion using SSRS as well as traditional methods, (b) evaluate the efficacy of targeted CD124+ cell depletion in chronic murine TB with and without anti-TB drug therapy, (c) assess the impact of dual CD25+ and CD124+ cell depletion in the TB granuloma with B6 sst1R and sst1S mice using SSRS as well as traditional methods, and (d) evaluate the efficacy of dual CD25+ and CD124+ cell depletion in chronic murine TB with and without anti-TB drug therapy. Aim 3: conduct translational applications of CD25+ cell depletion with (i) BCG vaccination and (ii) HIV reactivation. We will (a) evaluate the efficacy of targeted CD25+ cell depletion as a vaccine adjuvant prior to BCG vaccination and (b) evaluate the impact of targeted CD25+ cell depletion on HIV reactivation in PBMCs from patients with controlled HIV infection.

项目概要 Tregs 和 MDSC 已成为抑制结核肉芽肿中效应免疫的关键抑制细胞。 此外，Tregs 与 HIV 潜伏期的维持有关。我们生成了两个选择性的 使用白喉毒素 (DT) 融合蛋白有效消除 Tregs 和 MDSC 的生物制剂 技术：DT-IL2 靶向 IL2-R 阳性 (CD25+) 细胞 (Treg)，DT-IL4 靶向 IL4-R 阳性 (CD124+) 细胞（MDSC）。我们的初步数据表明：(i) DT-IL2 和 DT-IL4 降低了 Treg 和 Treg 的丰度。 分别在小鼠肺和脾中的 MDSC 细胞，(ii) DT-IL2 和 DT-IL4 降低了结核分枝杆菌的能力 在急性小鼠结核感染模型中增殖，(iii) 药物，他喹莫德，具有有效的抗 MDSC 活性， 降低了结核分枝杆菌在急性结核感染模型中的增殖能力，同时减少了 MDSC， (iv) 对 HIV 受控患者的 CD4+ T 细胞施用 DT-IL2 可重新激活病毒复制。 这些数据支持以下假设：(1) Tregs 和 MDSC 在结核分枝杆菌中发挥重要的抑制作用 (2) Treg 细胞在 HIV 潜伏期维持中发挥作用。这个的核心科学前提 应用是进一步定义 Tregs 和 MDSC 在 TB 和 HIV 感染中的免疫抑制作用 将揭示重要的免疫发病机制，可用于开发改进的 针对这两种疾病的宿主定向疗法（HDT）。在本提案中，我们试图通过追求来解决这个前提 以下目标。目标 1：利用靶向 CD25+ 细胞去除确定 Treg 细胞在结核肉芽肿中的作用 与 DT-IL2 一起。我们将 (a) 评估细胞（C57BL/6、WT，均为 sst1R）和 使用单细胞 RNA-Seq 检测有或没有 CD25+ 细胞耗竭的坏死性 (C57BL/6 sst1S) 小鼠肉芽肿 (SSRS) 以及传统方法，(b) 评估靶向 CD25+ 细胞耗竭在慢性疾病中的功效 接受和不接受抗结核药物治疗的小鼠结核病，以及 (c) 评估 Treg 细胞对该过程的贡献 使用兔空洞结核的新型模型来观察空化（肉芽肿破裂）。目标 2：定义角色 使用 DT-IL4 靶向去除 CD124+ 细胞，观察结核肉芽肿中的 MDSC。我们将 (a) 评估蜂窝 使用 SSRS 分析具有和不具有 CD124+ 细胞耗竭的细胞和坏死鼠肉芽肿的组成 以及传统方法，(b) 评估靶向 CD124+ 细胞去除对慢性小鼠结核病的疗效 有或没有抗结核药物治疗，(c) 评估双重 CD25+ 和 CD124+ 细胞消除对结核病的影响 使用 SSRS 以及传统方法对 B6 sst1R 和 sst1S 小鼠进行肉芽肿检测，以及 (d) 评估 双重 CD25+ 和 CD124+ 细胞消除对慢性小鼠结核病（有或没有抗结核药物治疗）的疗效。 目标 3：通过 (i) BCG 疫苗接种和 (ii) HIV 进行 CD25+ 细胞消除的转化应用 重新激活。我们将 (a) 在使用之前评估靶向 CD25+ 细胞去除作为疫苗佐剂的功效 BCG 疫苗接种和 (b) 评估靶向 CD25+ 细胞去除对 PBMC 中 HIV 重新激活的影响 来自艾滋病毒感染得到控制的患者。