Harnessing adaptive NK cell transfer to deplete viral reservoirs

利用适应性 NK 细胞转移来耗尽病毒库

• 批准号: 10393660
• 负责人: Edward Barker
• 金额: $ 75.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-16 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10393660
• 关键词: Activated Natural Killer Cell; Adoptive Transfer; Agonist; Allogenic; Animals; Antibodies; Antibody Specificity; Antibody-Dependent Enhancement; Autologous; Bar Codes; Binding; C Type Lectin Receptors; CD3 Antigens; Cell Therapy; Cells; Cellular Assay; Chronic; Control Animal; Coupling; Development; Disease; Disease Progression; Effector Cell; Engineering; FCGR3B gene; Fc Receptor; Genetic Polymorphism; Genotype; HIV; HIV-1; Human; Immunoglobulins; Individual; Infection; KLRD1 gene; Killer Cells; Ligands; Ligation; MHC Class I Genes; Macaca; Macaca mulatta; Memory; Molecular; NK cell therapy; Natural Killer Cells; Pathway interactions; Peptide/MHC Complex; Peptides; Persons; Plasma; Primates; Rapid screening; Receptor Cell; Rhesus; Role; SIV; Signal Transduction; Specificity; Surface; Testing; Viral; Viral Load result; Viral reservoir; Virus; Virus Replication; antibody-dependent cell cytotoxicity; antiretroviral therapy; cancer cell; cancer immunotherapy; experimental study; lymph nodes; receptor; response; tumor; viral rebound

PROJECT SUMMARY Natural killer (NK) cells provide an immediate defense against viruses and tumors by virtue of their ability to respond to infected or malignant cells without prior antigenic stimulation. This is accomplished through the integration of signals from activating and inhibitory NK cell receptors (aNKRs & iNKRs). In humans and other primate species, these include C-type lectin receptors, such as CD94/NKG2A and CD94/NKG2C, and the highly polymorphic killer-cell immunoglobulin-like receptors (KIRs), both of which interact with MHC class I ligands. These receptor-ligand interactions are fundamental to the ability of NK cells to differentiate healthy cells from unhealthy cells and provide a potential mechanism of specificity for the development of “NK cell memory”. NK cells can have a significant impact on HIV-1 infection. KIR and HLA class I polymorphisms have been identified that are associated with lower viral loads and slower courses of disease progression and certain NK cell subsets can kill HIV-infected cells in culture. Thus, NK cell-based therapies represent a promising approach for targeting HIV-infected cells and reducing the size of viral reservoirs. We hypothesize that viral peptides bound by the MHC class I ligands of aNKRs are critical to NK cell recognition and killing of HIV/SIV-infected cells and that the adoptive transfer of ex vivo activated NK cells in combination with latency reversal can deplete viral reservoirs in SIV-infected macaques on suppressive antiretroviral therapy (ART). In Aim 1, we will determine the contribution of viral peptides bound by MHC class I ligands of aNKRs to NK cell recognition of HIV- and SIV-infected cells. These studies will utilize high-throughput cellular assays to rapidly screen viral peptides for MHC class I interactions with aNKRs and to identify substitutions that disrupt these interactions. The corresponding changes will be introduced into HIV-1 and SIV to assess their impact on NK cell responses to virus-infected cells. In Aim 2, we will assess the capacity of ex vivo expanded NK cells in combination with latency reversal to deplete viral reservoirs in SIV-infected, ART-suppressed rhesus macaques. This aim will take advantage of barcoded SIV and a potent new latency reversal agent to compare with maximal sensitivity the ability of autologous versus allogeneic NK cell transfer to reduce the rate of viral reactivation after discontinuing ART. In Aim 3, we will test the hypothesis that the depletion of viral reservoirs by adaptive NK cell transfer can be enhanced by an Env-specific antibody with antibody-dependent cellular cytotoxicity against SIV-infected cells. This aim will use a similar approach as Aim 2 to determine the extent to which coupling NK cell effector function to the unparalleled specificity of antibodies can maximize reservoir depletion. These unprecedented studies will provide a better understanding of the role of viral peptides in NK cell recognition of HIV- and SIV-infected cells and an important proof-of-concept for the development of NK cell therapies to eradicate HIV-1 reservoirs in chronically infected individuals.

项目摘要 天然杀手（NK）细胞可以立即防御病毒和肿瘤，以致其能力 在没有事先抗原刺激的情况下应对感染或恶性细胞。这是通过 在人类和其他 灵长类动物，其中包括C型杠杆受体，例如CD94/NKG2A和CD94/NKG2C，以及 高度多态性杀手细胞免疫球蛋白样受体（KIRS），两者都与MHC I类相互作用 配体。这些受体 - 配体相互作用是NK细胞区分健康的能力的基础 细胞来自不健康的细胞，并为开发“ NK细胞的开发”提供了潜在的特异性机制 记忆”。NK细胞可能会对HIV-1感染产生重大影响。KIR和HLA I类多态性具有 被鉴定出与较低的病毒载荷和疾病进展和较慢的疾病进展过程相关的 某些NK细胞亚群可以杀死培养物中的HIV感染细胞。那就是NK基于细胞的疗法代表 靶向艾滋病毒感染细胞并减少病毒储层的大小的有前途的方法。我们假设 ANKR的MHC I类配体约束的病毒肽对于NK细胞识别和杀死至关重要 HIV/SIV感染的细胞，并且离体激活的NK细胞的自适应转移与潜伏期结合 在抑制性抗逆转录病毒疗法（ART）中，可以将逆转在SIV感染的猕猴中耗尽病毒库。 在AIM 1中，我们将确定由ANKR的MHC I类结合的病毒肽对NK的贡献 HIV和SIV感染细胞的细胞识别。这些研究将利用高通量的细胞测定 MHC I类与ANKRS相互作用的快速筛选病毒肽，并确定破坏的替代品 这些相互作用。相应的更改将引入HIV-1和SIV，以评估其对 NK细胞对病毒感染细胞的反应。在AIM 2中，我们将评估离体扩展的NK细胞的能力 结合潜伏期逆转以在SIV感染的Art抑制的恒河神中复制病毒储层 猕猴。这个目标将利用条形码的SIV和潜在的新延迟逆转代理来比较 具有最大灵敏度的自体和同种异体NK细胞转移的能力降低病毒率 中断艺术后的重新激活。在AIM 3中，我们将检验以下假设：病毒库的耗竭 通过具有抗体依赖性细胞的ENV特异性抗体可以增强自适应NK细胞转移 针对感染SIV的细胞的细胞毒性。此目标将使用类似的方法与目标2确定 哪种将NK细胞效应子函数耦合到无与伦比的抗体特异性可以最大化储层 消耗。这些前所未有的研究将更好地理解病毒肽在NK中的作用 HIV和SIV感染细胞的细胞识别以及NK细胞发展的重要概念验证 在长期感染的个体中对放射性HIV-1储存剂进行疗法。