HIV eradication by ADCC-activated NK cell killing

通过 ADCC 激活的 NK 细胞杀伤来根除 HIV

基本信息

批准号：
9243206
负责人：
JONATHAN KARN
金额：
$ 46.94万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-15 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9243206
关键词：
Acquired Immunodeficiency Syndrome Activated Natural Killer Cell Acute Affinity Anti-Retroviral Agents Antibodies Antigen Targeting Antiviral Agents Autologous Biological Assay CD4 Positive T Lymphocytes Caring Cell model Cells Chronic Clinic Clinical Research Cytokine Activation DNA Data Down-Regulation Drug Costs Effectiveness Ensure Epitopes Flow Cytometry Genetic Transcription Goals HIV HIV Infections HIV-1 Highly Active Antiretroviral Therapy Histocompatibility Antigens Class I Histone Deacetylase Inhibitor Human Immune response Immune system Immunization Immunology Individual Infection Interruption Killer Cells Lead Life Ligands MHC Class I Genes Macaca mulatta Malignant Neoplasms Measurement Measures Mediating Medical Messenger RNA Microglia Modeling NK Cell Activation Natural Killer Cells Patients Peripheral Pharmaceutical Preparations Pharmacotherapy Phase Population Protocols documentation Proviruses Reaction Recruitment Activity Regulatory T-Lymphocyte Rest SIV Sampling Specificity T memory cell T-Lymphocyte Techniques Testing Thailand United States Use Effectiveness Vaccine Clinical Trial Viral reservoir Virus Virus Latency Virus Replication Vorinostat antibody-dependent cell cytotoxicity antiretroviral therapy base cell killing clinical investigation compare effectiveness design experimental study human subject in vivo killings macrophage memory CD4 T lymphocyte neutralizing antibody next generation sequencing polarized cell public health relevance receptor research clinical testing scale up simian human immunodeficiency virus viral rebound

项目摘要

DESCRIPTION (provided by applicant): In order to achieve a functional cure for HIV infections the size of the latent viral reservoir must be sufficiently reduced to allow an indefinie cessation of antiviral drug treatment. Our strategy combines a variety of approaches that have been successfully tested against cancer and HIV-1 with the goal of achieving a significant reduction of HIV reservoirs in patients who are well suppressed on HAART. Specifically, we will combine ex vivo cytokine activation of natural killer (NK) cells with stimulation of antibody-dependent cell-mediated cytotoxicity (ADCC) using broadly neutralizing antibodies against HIV-1 Env. Env expression will be achieved by induction of proviral transcription with latency reversal drugs, such as HDAC inhibitors that are currently undergoing clinical evaluation. During the R21 phase of this application we will use newly developed ex vivo models of HIV latency to optimize the strategy. In Specific Aim 1, we will define and optimize the specificity of NK cell-mediated killing of different CD4+ T cell subsets. This will involve measuring NK cell-activating/-inhibiting receptor ligands expressed on CD4+ T cell subsets using different latency-reversing drugs, measuring the efficacy of autologous NK cells to kill latently infected CD4+ T cell subsets, and optimizing ex vivo cytokine activation of NK cells. In Specific Aim 2, we will investigate the effectiveness of using broadly neutralizing antibodies against Env to stimulate ADCC in NK cell-mediated killing of latently HIV-1-infected primary T cells, in combination with proviral reactivation. Upon completion of the R21 phase of this project we will have established reliable assays to measure NK-mediated killing of reactivated latently infected T cells, defined protocols for activating NK cells ex vivo, and identified antibodies for use in ADCC-mediated killing. The R33 phase of this application will focus exclusively on ex vivo studies using patient cells that are designed to show elimination of latently infected cells. Human subjects will be recruited from our HIV clinic, the Special Immunology Unit (SIU, UHCMC). The SIU has an active population of 1026 HIV-infected patients who are followed regularly for routine medical care; approximately 90% are receiving antiretroviral therapy (ART). In Specific Aim 3, we will validate whether techniques during the R21 phase can enhance the killing of autologous primary T cells from HIV-infected individuals. In Specific Aim 4, we will assess whether techniques developed in aims 1, 2, and 3 also lead to a reduction in the latent endogenous viral reservoir in HIV-infected patients. We will perform pairwise comparisons of NK-killing in the presence and absence of antibody for ADCC, with and without T-cell induction and with and without NK cell activation. At least 10 patient samples will be analyzed to ensure statistical significance of the results. The readout for the assays will be quantitative HIV proviral DNA and mRNA measurements from quantitative PCR reactions analyzed by next-generation sequencing. The experiments in this proposal will set the stage for clinical studies of NK-directed eradication of latent HIV. Because our strategy combines enhancing NK selectivity by ADCC using available antibodies, ex vivo activation of NK cells and in vivo activation of latent proviruses, we believe it represents a practical solution that can be readily implemented in the clinic for a large number of patients.

描述（由申请人提供）：为了实现 HIV 感染的功能性治愈，必须充分减小潜伏病毒库的大小，以允许无限期停止抗病毒药物治疗。我们的策略结合了多种已成功针对癌症和 HIV-1 进行测试的方法，目标是显着减少接受 HAART 治疗后得到良好抑制的患者中的 HIV 储存库。具体来说，我们将使用针对 HIV-1 Env 的广泛中和抗体，将自然杀伤 (NK) 细胞的离体细胞因子激活与抗体依赖性细胞介导的细胞毒性 (ADCC) 的刺激结合起来。 Env 表达将通过潜伏逆转药物（例如目前正在进行临床评估的 HDAC 抑制剂）诱导原病毒转录来实现。在该应用的 R21 阶段，我们将使用新开发的 HIV 潜伏期离体模型来优化策略。在具体目标 1 中，我们将定义并优化 NK 细胞介导的不同 CD4+ T 细胞亚群的杀伤。这将涉及使用不同的潜伏期逆转药物测量 CD4+ T 细胞亚群上表达的 NK 细胞激活/抑制受体配体，测量自体 NK 细胞杀死潜伏感染的 CD4+ T 细胞亚群的功效，以及优化 NK 的离体细胞因子激活细胞。在具体目标 2 中，我们将研究使用针对 Env 的广泛中和抗体与前病毒再激活相结合，在 NK 细胞介导的杀伤潜伏 HIV-1 感染的原代 T 细胞中刺激 ADCC 的有效性。该项目的 R21 阶段完成后，我们将建立可靠的测定方法来测量 NK 介导的对重新激活的潜伏感染 T 细胞的杀伤，定义离体激活 NK 细胞的方案，并确定用于 ADCC 介导杀伤的抗体。该应用的 R33 阶段将专门专注于使用患者细胞进行离体研究，这些细胞旨在显示潜伏感染细胞的消除。人类受试者将从我们的 HIV 诊所、特殊免疫学单位 (SIU、UHCMC) 招募。 SIU 拥有 1026 名 HIV 感染者，他们定期接受常规医疗护理；大约 90% 正在接受抗逆转录病毒治疗 (ART)。在具体目标 3 中，我们将验证 R21 阶段的技术是否可以增强对 HIV 感染者的自体原代 T 细胞的杀伤。在具体目标 4 中，我们将评估目标 1、2 和 3 中开发的技术是否也能减少 HIV 感染患者中潜在的内源性病毒库。我们将在存在和不存在 ADCC 抗体、有和没有 T 细胞诱导以及有和没有 NK 细胞激活的情况下进行 NK 杀伤的成对比较。将分析至少 10 个患者样本，以确保结果的统计显着性。测定的读数将是通过下一代测序分析的定量 PCR 反应的定量 HIV 前病毒 DNA 和 mRNA 测量结果。该提案中的实验将为 NK 引导根除潜伏 HIV 的临床研究奠定基础。因为我们的策略结合了使用可用抗体通过 ADCC 增强 NK 选择性、NK 细胞的离体激活和潜伏原病毒的体内激活，所以我们相信它代表了一种实用的解决方案，可以在临床上为大量患者轻松实施。