Generation of highly differentiated NK cells to act in synergy with broadly neutralizing antibodies to reduce the SIV reservoir and establish viral control in absence of ART

生成高度分化的 NK 细胞，与广泛中和抗体协同作用，减少 SIV 储存库，并在没有 ART 的情况下建立病毒控制

• 批准号: 10883005
• 负责人: Mirko Paiardini
• 金额: $ 81.28万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-22 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10883005
• 关键词: Abnormal Cell; African Green Monkey; Animals; Antibody Therapy; Antibody-Dependent Enhancement; Automobile Driving; B-Lymphocytes; Biological Assay; Blood; CD8B1 gene; Cells; Collaborations; Complex; Coupled; DNA; Data; Development; Disease remission; Drug or chemical Tissue Distribution; Education; Effector Cell; Environment; Exhibits; FCGR3B gene; Fc Receptor; Frequencies; Gastrointestinal tract structure; Generations; Goals; HIV; Immune; Immunotherapy; In Situ; Infection; Inflammation; Inflammatory; Intercept; Interruption; Intervention; Kinetics; Lymphoid; Lymphoid Tissue; Macaca mulatta; Maintenance; Measures; Mediating; Modeling; Natural Killer Cells; Oncology; Pathogenicity; Peptides; Persons; Phenotype; Positioning Attribute; Pre-Clinical Model; Production; Proliferating; Property; Recording of previous events; Reporting; Research Personnel; SIV; T-Lymphocyte; Testing; Time; Tissues; Viral; Viral Load result; Viral Physiology; Viral reservoir; Virus; Virus Diseases; Virus Replication; Withholding Treatment; Work; antibody-dependent cell cytotoxicity; antiretroviral therapy; cell killing; cohort; cytokine; cytotoxic; cytotoxicity; design; experience; genome-wide; in vivo; innovation; insight; interleukin-21; interleukin-21 receptor; lymph nodes; neutralizing antibody; neutralizing vaccine; neutrophil; preservation; prevent; receptor; response; synergism; transcriptome; transcriptome sequencing; vaccine efficacy; viral rebound; virus host interaction

PROJECT SUMMARY/ABSTRACT Lymph nodes and the gastrointestinal tract are major tissue viral reservoirs in people living with HIV and treated with antiretroviral treatment (ART). A distinguishing feature of SIV infection in African green monkeys (AGMs), a natural host species for SIV, is the absence of viral dissemination in the lymphoid B-cell follicle (BCF) and the presence of strong NKcell-mediated control of viral replication in the T-cell zone and BCF of secondary lymphoid tissues (SLT). In SIVagm-infected AGMs, we recently identified the expansion of NK cells with a terminally-differentiated phenotype expressing (i) high levels of CD16, an activating Fc receptor that mediates antibody-dependent cellular cytotoxicity (ADCC), and (ii) low levels of NKG2a, an inhibitory receptor, that modulates NK cell education via interactions with MHC-E. This terminally-differentiated NK cell (NKTD) subset (NKG2alowCD16+) exhibited adaptive-like properties and, due at least in part to reduced NKG2a expression, showed high cytolytic activity against target cells presenting SIV-Env peptides in the context of MHC-E. The formation of adaptive NKTD cells was blocked in SIVmac-infected rhesus macaques (RMs) in favor of a subset (NKG2ahiCD16+) with pro-inflammatory function, such as production of IFN, a cytokine also known for its capacity to increase MHC-E expression on target cells. Remarkably, in a separate cohort of SIVmac-infected, ART-treated RMs, we demonstrated that AGM-like profiles of adaptive NKTD cells are rescued via treatment with interleukin-21 (IL-21). In IL-21 treated RMs, the frequency and responsiveness of the adaptive NKTD cells to target cells presenting SIV peptides in the context of MHC-E strongly correlated with a reduction of SIV DNA in the gut, lower levels of replication competent virus in SLT, and a delay in viral rebound following ART interruption. In Aim 1 of this proposal we will define (I) the mechanisms driving the formation and activity of adaptive NKTD cells, such as cytokine environment and lymphoid inflammation; (II) their tissue distribution; and (III) ADCC capacity. In addition to NK cells, the impact of IL-21 will be investigated in T-cells and other immune cells that can respond to IL-21. In Aim 2, we will assess whether and to what extent the combination of IL-21-induced NKTD cells and a cocktail of anti-SIV broadly neutralizing antibodies (bNAbs) facilitates, through ADCC-mediated clearance of infected cells, a reduction of viral burden and control of viral rebound following ART interruption. Specifically, in SIVmac-infected, ART-treated RMs, the “differentiate, target, and kill” approach will allow differentiation of the NKTD cells with IL-21 followed by targeting of cells harboring reactivated virus with bNAbs. Using a model reproducing the complex virus-host interactions occurring in people living with HIV, we expect that the rescue of NK cell terminal differentiation will promote robust ADCC activity, which will synergize with the selective targeting of viral reservoirs that reactivate following ATI, thus leading to prolonged ART-free remission.

项目概要/摘要 淋巴结和胃肠道是艾滋病毒感染者的主要组织病毒库 非洲绿猴感染 SIV 的一个显着特征是接受抗逆转录病毒治疗 (ART)。 (AGM) 是 SIV 的天然宿主物种，是指淋巴 B 细胞滤泡中不存在病毒传播 (BCF) 以及 T 细胞区和 BCF 中存在强大的 NK 细胞介导的病毒复制控制 在 SIVagm 感染的 AGM 中，我们最近发现了 NK 细胞的扩增。 具有终末分化表型表达 (i) 高水平的 CD16，一种激活 Fc 受体 介导抗体依赖性细胞毒性 (ADCC)，以及 (ii) 低水平的 NKG2a（一种抑制性受体）， 通过与 MHC-E 这种终末分化的 NK 细胞 (NKTD) 相互作用来调节 NK 细胞的教育。 子集 (NKG2alowCD16+) 表现出类似适应性的特性，并且至少部分归因于 NKG2a 的减少 表达，对呈递 SIV-Env 肽的靶细胞表现出高细胞溶解活性 MHC-E 在 SIVmac 感染的恒河猴 (RM) 中适应性 NKTD 细胞的形成被阻断 有利于具有促炎功能的子集（NKG2ahiCD16+），例如产生 IFNα（也是一种细胞因子） 值得注意的是，在一个单独的群体中，它具有增加靶细胞 MHC-E 表达的能力。 SIVmac 感染、ART 处理的 RM，我们证明了适应性 NKTD 细胞的 AGM 样特征被拯救 通过用白细胞介素 21 (IL-21) 治疗，在 IL-21 治疗的 RM 中，IL-21 的频率和反应性。 NKTD 细胞对在 MHC-E 背景下呈递 SIV 肽的靶细胞的适应性与 肠道中 SIV DNA 减少、SLT 中具有复制能力的病毒水平降低以及病毒反弹延迟 在 ART 中断之后，在本提案的目标 1 中，我们将定义 (I) 驱动形成的机制。 适应性 NKTD 细胞的活性，例如细胞因子环境和淋巴炎症 (II) 其组织； (III) ADCC 能力 除了 NK 细胞外，还将研究 IL-21 对 T 细胞的影响。 以及其他可以对 IL-21 做出反应的免疫细胞 在目标 2 中，我们将评估是否以及程度如何。 IL-21 诱导的 NKTD 细胞与抗 SIV 广泛中和抗体 (bNAb) 混合物的组合 通过 ADCC 介导的感染细胞清除，促进减少病毒负荷并控制病毒 具体来说，在感染 SIVmac 并接受 ART 治疗的 RM 中，“区分、 “靶向并杀死”方法将允许使用 IL-21 分化 NKTD 细胞，然后靶向细胞 使用 bNAb 携带重新激活的病毒，使用重现复杂病毒-宿主相互作用的模型。 发生在 HIV 感染者身上，我们预计 NK 细胞终末分化的拯救将促进 强大的 ADCC 活性，这将与重新激活的病毒库的选择性靶向产生协同作用 ATI 后，从而导致长期无 ART 缓解。