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.

项目摘要 TREG和MDSC已成为抑制TB肉芽肿的效应免疫的关键抑制细胞。 此外，Tregs与维持艾滋病毒潜伏期有关。我们已经产生了两个选择性 使用白喉毒素（DT）融合蛋白的Tregs和MDSC有效耗竭的生物学剂 技术：DT-IL2靶向IL2-R阳性（CD25+）细胞（Tregs）和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减少了Treg和 MDSC细胞分别在小鼠肺和脾脏中，（ii）DT-IL2和DT-IL4降低了M. tb的能力 在急性鼠结核病感染模型中增殖，（iii）具有有效抗MDSC活性的药物，tasquinimod， M. TB在急性结核病感染模型中增殖的能力降低了MDSC的降低， （iv）来自受控HIV患者的DT-IL2对CD4+ T细胞的施用可重新激活病毒复制。 这些数据支持（1）Treg和MDSC在M. TB中起重要抑制作用的假设。 遏制和（2）Treg细胞在HIV潜伏期维持中起作用。一个中心科学前提 应用是TB和HIV感染中Treg和MDSC的免疫抑制作用的进一步定义 将揭示重要的免疫原理机制，这些机制可能被利用用于发展的改善 两种疾病的宿主定向疗法（HDTS）。在此提案中，我们寻求通过追求来解决这一前提 以下目标。 AIM 1：使用靶向的CD25+细胞耗竭来定义Treg细胞在TB肉芽肿中的作用 与DT-IL2。我们将（a）评估细胞组成的细胞组成（C57BL/6，wt是SST1R）和 坏死（C57BL/6 SST1S）使用单细胞RNA-Seq进行有或没有CD25+细胞耗竭的鼠颗粒 （SSR）以及传统方法，（b）评估靶向CD25+细胞耗竭在慢性中的功效 有或没有抗TB药物治疗的鼠结核 使用新型兔腔TB模型的空化（肉芽肿分解）。目标2：定义角色 使用靶向的CD124+细胞用DT-IL4进行靶向的CD124+细胞耗竭，MDSC在TB肉芽肿中。我们将（a）评估细胞 使用有或没有CD124+细胞耗尽的细胞和坏死鼠颗粒的组成 以及传统方法，（b）评估靶向CD124+细胞耗竭在慢性鼠TB中的疗效 在有或没有抗TB药物治疗的情况下，（c）评估双CD25+和CD124+细胞在结核病中的影响 使用SSR和传统方法的B6 SST1R和SST1S小鼠的肉芽肿，（D）评估 双重CD25+和CD124+细胞耗竭在慢性鼠结核病中有和没有抗TB药物治疗的功效。 AIM 3：通过（I）BCG疫苗接种和（II）HIV进行CD25+细胞耗竭的翻译应用 重新激活。我们将（a）评估靶向CD25+细胞耗竭作为疫苗佐剂的疗效 BCG疫苗接种和（b）评估靶向CD25+细胞耗竭对PBMC中HIV重新激活的影响 来自受控HIV感染的患者。