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.

项目总结/摘要 淋巴结和胃肠道是HIV感染者的主要组织病毒库， 抗逆转录病毒治疗（ART）。非洲绿色猴SIV感染的一个显著特征 （AGM）是SIV的天然宿主物种，其特征在于在淋巴B细胞滤泡中没有病毒传播 (BCF)以及NK细胞介导的对T细胞区和BCF中病毒复制的强控制的存在， 次级淋巴样组织（secondary lymphoid tissues，ELT）。在SIVagm感染的AGM中，我们最近发现NK细胞的扩增， 具有表达（i）高水平的CD 16的终末分化表型，CD 16是一种活化Fc受体， 介导抗体依赖性细胞毒性（ADCC），和（ii）低水平的NKG 2a，一种抑制性受体， 通过与MHC-E的相互作用调节NK细胞教育。这种终末分化的NK细胞（NKTD） 亚群（NKG 2alowCD 16+）表现出适应性样特性，至少部分由于NKG 2a减少 表达，显示出高的细胞溶解活性对靶细胞呈递SIV-Env肽的背景下， 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亚群（NKG 2ahiCD 16+），如产生IFN γ，一种细胞因子也 以其增加靶细胞上MHC-E表达的能力而闻名。值得注意的是，在一个单独的队列， SIVmac感染，ART治疗的RM，我们证明了自适应NKTD细胞的AGM样特征被拯救， 白细胞介素-21（IL-21）治疗。在IL-21处理的RM中， 在MHC-E的背景下，NKTD细胞对呈递SIV肽的靶细胞的适应性与NKTD细胞对SIV肽的耐受性强相关。 肠道中SIV DNA的减少，肠道中具有复制能力的病毒的水平降低，以及病毒反弹的延迟 ART中断后。在本提案的目标1中，我们将定义（I）驱动形成的机制 和适应性NKTD细胞的活性，如细胞因子环境和淋巴炎症;（II）它们的组织 （3）ADCC能力。除NK细胞外，还将在T细胞中研究IL-21的影响 以及其他免疫细胞对IL-21的反应。在目标2中，我们将评估 IL-21诱导的NKTD细胞和抗SIV广泛中和抗体（bNAb）混合物的组合 通过ADCC介导的感染细胞的清除，有助于减少病毒负荷和控制病毒感染。 抗逆转录病毒疗法中断后反弹。具体而言，在SIVmac感染的ART治疗的RM中，“分化， 靶向和杀伤”方法将允许用IL-21分化NKTD细胞，然后靶向细胞 携带带有bNAb的再活化病毒。使用一个模型再现复杂的病毒-宿主相互作用 发生在艾滋病毒感染者中，我们预计拯救NK细胞终末分化将促进 强大的ADCC活性，这将与选择性靶向病毒储库协同作用， ATI后，从而导致延长的无ART缓解。