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：
8656258
负责人：
JONATHAN KARN
金额：
$ 21.79万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-12-01 至 2015-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8656258
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 Clinical Trials Cytokine Activation DNA Data Down-Regulation 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 Immunology Individual Infection Interruption 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 Solutions Specificity Staging T memory cell T-Lymphocyte T-Lymphocyte Subsets Techniques Testing Thailand United States Use Effectiveness Vaccine Clinical Trial Viral Virus Virus Replication Vorinostat antibody-dependent cell cytotoxicity antiretroviral therapy base cell killing compare effectiveness design human subject in vivo killings macrophage memory CD4 T lymphocyte neutralizing antibody next generation sequencing public health relevance receptor research clinical testing research study scale up simian human immunodeficiency virus

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）：为了实现艾滋病毒感染的功能治愈，必须充分降低潜在病毒储存剂的大小，以允许对抗病毒药物治疗的不确定。我们的策略结合了已成功针对癌症和HIV-1测试的各种方法，目的是在HAART受到良好抑制的患者中大大降低了HIV储层。具体而言，我们将使用针对HIV-1 Env的抗体依赖性细胞介导的细胞介导的细胞介导的细胞毒性（ADCC）结合体内细胞因子激活（NK）细胞的活化。 ENV表达将通过使用潜伏期逆转药物（例如目前正在进行临床评估的HDAC抑制剂）来实现。在此应用程序的R21阶段，我们将使用新开发的艾滋病毒潜伏期的离体模型来优化策略。在特定目标1中，我们将定义和优化 NK细胞介导的不同CD4+ T细胞子集的杀伤。这将涉及测量使用不同的逆向药物在CD4+ T细胞子集上表达的NK细胞激活/抑制受体配体，测量自体NK细胞的疗效，杀死了潜在的受感染的CD4+ T细胞亚群，并优化了NK细胞的EXTokine EXTOKINE活化。在特定的目标2中，我们将研究使用针对ENV的广泛中和抗体刺激NK细胞介导的LAIV抗HIV-1感染的原代T细胞的ADCC的有效性，并结合前病毒的重新激活。该项目的R21阶段完成后，我们将建立可靠的测定法，以测量NK介导的杀死被重新激活的T细胞的杀死，定义了用于激活NK细胞的方案，并确定了用于ADCC介导的杀伤的抗体。该应用程序的R33相将专门集中于使用旨在表现出消除潜在感染细胞的患者细胞的离体研究。人类受试者将从我们的艾滋病毒诊所（SIU，UHCMC）中招募。 SIU的活跃人群为1026名受HIV感染的患者，经常受到常规医疗服务；大约90％正在接受抗逆转录病毒疗法（ART）。在特定的目标3中，我们将验证R21期间的技术是否可以增强受HIV感染者自体原代细胞的杀死。在特定目标4中，我们将评估目标1、2和3中的技术是否还会导致HIV感染患者中潜在的内源性病毒储存剂的降低。在存在和没有T细胞诱导的ADCC抗体的存在和不存在抗体的情况下，我们将对NK杀死的成对比较，并且有或没有NK细胞活化。将至少分析10个患者样本，以确保结果的统计显着性。该测定的读数将是定量HIV病毒DNA和通过下一代测序分析的定量PCR反应的mRNA测量值。该提案中的实验将为NK指导根除潜在艾滋病毒的临床研究奠定了基础。由于我们的策略结合了通过可用抗体，通过可用的NK细胞激活和在体内激活潜在的预科病毒的体内激活来增强NK的选择性，因此我们相信它代表了一种实用的解决方案，可以在诊所中为大量患者提供很容易实施的解决方案。