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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要自然杀伤 (NK) 细胞能够立即防御病毒和肿瘤在没有事先抗原刺激的情况下对感染或恶性细胞做出反应这是通过整合人类和其他细胞中激活和抑制性 NK 细胞受体（aNKR 和 iNKR）的信号。灵长类动物，包括 C 型凝集素受体，例如 CD94/NKG2A 和 CD94/NKG2C，以及高度多态性的杀伤细胞免疫球蛋白样受体 (KIR)，两者均与 I 类 MHC 相互作用这些受体-配体相互作用对于 NK 细胞分化健康细胞的能力至关重要。从不健康细胞中提取细胞，并为“NK细胞”的发育提供潜在的特异性机制 NK 细胞可对 HIV-1 感染产生重大影响。KIR 和 HLA I 类多态性具有显着影响。已被确定与较低的病毒载量和较慢的疾病进展有关某些 NK 细胞亚群可以杀死培养中的 HIV 感染细胞，因此，基于 NK 细胞的疗法代表了一种治疗方法。针对 HIV 感染细胞并减少病毒库大小的有前途的方法。与 aNKR 的 MHC I 类配体结合的病毒肽对于 NK 细胞识别和杀伤至关重要 HIV/SIV 感染细胞以及离体激活的 NK 细胞的过继转移与潜伏期相结合在接受抑制性抗逆转录病毒治疗（ART）的感染 SIV 的猕猴中，逆转可以耗尽病毒库。在目标 1 中，我们将确定 aNKR 的 MHC I 类配体结合的病毒肽对 NK 的贡献这些研究将利用高通量细胞检测来识别 HIV 和 SIV 感染的细胞。快速筛选病毒肽的 MHC I 类与 aNKR 的相互作用，并识别破坏的替代这些相互作用将被引入到 HIV-1 和 SIV 中，以评估它们对病毒的影响。 NK 细胞对病毒感染细胞的反应在目标 2 中，我们将评估离体扩增的 NK 细胞的能力。与潜伏期逆转相结合，以耗尽 SIV 感染、ART 抑制的恒河猴中的病毒库该目标将利用带条形码的 SIV 和一种有效的新型潜伏期逆转剂进行比较。以最大的灵敏度，自体与同种异体 NK 细胞转移能够降低病毒感染率停止 ART 后重新激活在目标 3 中，我们将检验病毒库耗尽的假设。通过适应性 NK 细胞转移，可以通过具有抗体依赖性细胞的 Env 特异性抗体来增强针对 SIV 感染细胞的细胞毒性该目标将使用与目标 2 类似的方法来确定细胞毒性的程度。将 NK 细胞效应功能与抗体无与伦比的特异性相结合，可以最大限度地提高储存库这些史无前例的研究将有助于更好地了解病毒肽在 NK 中的作用。 HIV 和 SIV 感染细胞的细胞识别以及 NK 细胞发育的重要概念验证根除慢性感染者中 HIV-1 病毒库的疗法。