Optimizing the Depletion of innate immune Effector Cells in Nonhuman Primates

优化非人灵长类动物先天免疫效应细胞的消耗

• 批准号: 10646290
• 负责人: Afamefuna Okoye
• 金额: $ 20.54万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10646290
• 关键词: Acute; Adaptive Immune System; Antibodies; Antigens; Autologous; B-Lymphocytes; Benchmarking; Binding; Biological Assay; Biology; Blood; Bone Marrow; CCR5 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell surface; Cells; Chronic; Complex; DNA; Data; Dose; Effector Cell; Engineering; FCGR3B gene; Future; Goals; HIV; HIV Infections; HIV vaccine; Herpesviridae; Human; Human Herpesvirus 8; Immune; Immune response; Immunologic Markers; In Vitro; Infusion procedures; Innate Immune Response; Innate Immune System; Interleukin-15; JAK3 gene; Lead; Lentivirus Vector; Leukapheresis; Liver; Lymphocyte; Lymphoid Tissue; MS4A1 gene; Macaca mulatta; Macrophage; Mediating; Modality; Modeling; Monoclonal Antibodies; Myelogenous; Myeloid Cells; Natural Immunity; Natural Killer Cells; Oncogenic; Pathogenesis; Performance; Play; Population; Proliferating; Prophylactic treatment; Quality Control; Rhadinovirus; Rhesus; Role; SIV; Series; Signal Pathway; Signal Transduction; Specificity; Spleen; T memory cell; T-Lymphocyte; Techniques; Testing; Time; Tissues; Validation; Virus; Virus Diseases; Virus Replication; acute infection; anti-CD20; antibody-dependent cell cytotoxicity; antiviral immunity; cell type; chimeric antigen receptor; chimeric antigen receptor T cells; chronic infection; cytotoxic; design; experimental study; gammaherpesvirus; granulocyte; in vivo; inhibitor; innovation; interest; loss of function; lymph nodes; manufacture; monocyte; nonhuman primate; novel; peripheral blood; response; vaccine development

PROJECT SUMMARY Innate immunity plays an important role in the host response against HIV infection. Crucial to this response are innate immune effector cells including NK cells, monocytes, macrophages and granulocytes, which possess numerous anti-HIV activities. Notably, monocytes and macrophages may also contribute to HIV persistence during suppressive therapy by harboring integrated copies of replication-competent HIV proviral DNA. Cytotoxic NK cells and other Fcγ-RIIIA (CD16) expressing cells can bind to specific antibodies to enable antibody- dependent cellular cytotoxicity (ADCC), which can mediate killing of HIV-infected cells. However, the nonredundant role of innate immune effector activity is often difficult to define in vivo. A key barrier is that modalities to deplete these innate immune cells for loss-of-function studies (e.g., Janus kinase 3 inhibitors, anti- IL-15 monoclonal antibodies [mAb], anti-MHC-II mAb) can also impact cells of the adaptive immune system, namely T cells. For example, we previously used anti-IL-15 mAbs to neutralize IL-15 signaling in vivo in rhesus macaques (RM) to assess the role of NK cells during acute and chronic infection, using the CCR5-tropic SIVmac239. We observed a massive depletion of NK cells in blood and tissues (DeGotarrdi et al., J. Immunol. 2016), surprisingly minimal impact on SIV replication, and a significant reactivation of oncogenic gammaherpes viruses (Okoye et al., J. Immunol. 2019). In particular, we noted a reactivation of rhesus RM rhadinovirus (RRV), a simian γ-herpesvirus closely related to human herpesvirus type 8/Kaposi's sarcoma–associated herpesvirus, which may have been directly related to the depletion of NK cells, or alternatively due to the impact of IL-15 signaling blockade on other lymphocyte populations such as CD4+ and CD8+ effector memory T cells. Likewise, use of an anti-CD16 depleting mAb to assess the role of CD16/Fcγ-RIIIA in SIV-infected RM resulted in transient and often incomplete loss-of-function (Choi et al., J. Virol. 2008). The experiments proposed here will overcome these limitations by using chimeric antigen receptor (CAR) T cells designed to deplete NKG2A+ and CD16+ cells in RM. Our preliminary data clearly shows our expertise in this approach using CD20-directed CAR T cells to deplete B cells in blood and lymphoid tissues of RM. We will use similar techniques to optimize two CAR molecules, which we have already designed and preliminarily validated. Each is designed to target markers that are expressed on the vast majority of NK cells in RM (NKG2A) or cells that can mediate ADCC activity (CD16), which will in turn induce their specific functional deficiency in vivo. CD16 CAR T cells may provide added value as an approach to deplete myeloid reservoirs. If successful, this project has the potential to substantially advance the utility of nonhuman primate models to study the specific role of innate immune effector activity in HIV pathogenesis, vaccine development and cure strategies.

项目概要 先天免疫在宿主对抗艾滋病毒感染的反应中发挥着重要作用。此响应的关键是 先天性免疫效应细胞，包括 NK 细胞、单核细胞、巨噬细胞和粒细胞，它们具有 许多抗艾滋病毒活动。值得注意的是，单核细胞和巨噬细胞也可能导致艾滋病毒持续存在 在抑制治疗期间，通过携带具有复制能力的 HIV 前病毒 DNA 的整合副本。细胞毒性 NK 细胞和其他 Fcγ-RIIIA (CD16) 表达细胞可以与特定抗体结合，使抗体- 依赖性细胞毒性（ADCC），可以介导杀死 HIV 感染细胞。然而， 先天免疫效应子活性的非冗余作用通常很难在体内定义。一个关键障碍是 消除这些先天免疫细胞以进行功能丧失研究的方式（例如 Janus 激酶 3 抑制剂、抗 IL-15 单克隆抗体 [mAb]、抗 MHC-II mAb）也可以影响适应性免疫系统的细胞， 即T细胞。例如，我们之前使用抗 IL-15 mAb 来中和恒河猴体内的 IL-15 信号传导 猕猴 (RM) 使用 CCR5-tropic 评估 NK 细胞在急性和慢性感染期间的作用 SIVmac239。我们观察到血液和组织中 NK 细胞大量减少（DeGotarrdi 等人，J.Immunol. 2016），令人惊讶的是对 SIV 复制的影响极小，并且致癌性伽马疱疹显着重新激活 病毒（Okoye 等人，J.Immunol.2019）。特别是，我们注意到恒河猴 RM 病毒 (RRV) 的重新激活， 与人类疱疹病毒 8 型/卡波西肉瘤相关疱疹病毒密切相关的猿猴 γ-疱疹病毒， 这可能与 NK 细胞的消耗直接相关，或者是由于 IL-15 的影响 对其他淋巴细胞群（如 CD4+ 和 CD8+ 效应记忆 T 细胞）的信号传导阻断。同样地， 使用抗 CD16 耗竭单克隆抗体评估 CD16/Fcγ-RIIIA 在 SIV 感染 RM 中的作用，导致短暂的 并且常常不完全丧失功能（Choi et al., J. Virol. 2008）。这里提出的实验将克服 通过使用旨在消除 NKG2A+ 和 CD16+ 细胞的嵌合抗原受体 (CAR) T 细胞来克服这些限制 单位：RM。我们的初步数据清楚地表明了我们在使用 CD20 定向 CAR T 细胞的方法方面的专业知识 消耗 RM 血液和淋巴组织中的 B 细胞。我们将使用类似的技术来优化两个 CAR 分子，我们已经设计并初步验证了。每个都旨在针对以下标记： 在 RM 中的绝大多数 NK 细胞 (NKG2A) 或可介导 ADCC 活性的细胞 (CD16) 上表达， 这反过来又会导致它们体内特定的功能缺陷。 CD16 CAR T 细胞可提供附加值 作为耗尽骨髓库的一种方法。如果成功，该项目有可能大幅推进 利用非人类灵长类动物模型研究先天免疫效应子活性在艾滋病毒中的具体作用 发病机制、疫苗开发和治疗策略。