Cell Surface Marker Combinations to Identify Latently Infected CD4+ Cells In Vivo

用于识别体内潜伏感染 CD4 细胞的细胞表面标记组合

• 批准号: 8719846
• 负责人: Fabio Romerio
• 金额: $ 40.03万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-12 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8719846
• 关键词: CD4 Positive T Lymphocytes; CD6 antigen; Cell surface; Cells; Clinical; Florida; Flow Cytometry; Flushing; Funding; Gene Expression Profile; Goals; Grant; HIV-1; Highly Active Antiretroviral Therapy; Immune response; Immunologic Surveillance; In Vitro; Infection; Investigation; Laboratories; Lead; Measures; Messenger RNA; Modeling; Molecular Profiling; Monitor; Outcome Study; Patients; Pharmaceutical Preparations; Proviruses; Refractory; Rest; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Shock; Specificity; Surface; System; T-Cell Activation; Techniques; Testing; United States National Institutes of Health; Validation; Viral; Virion; Virus; Virus Diseases; Work; antiretroviral therapy; cell type; in vitro Model; in vivo; killings; lymph nodes; memory CD4 T lymphocyte; novel therapeutics; peripheral blood; prevent; public health relevance; receptor; therapeutic target; therapy development; viral RNA

DESCRIPTION (provided by applicant): A major hurdle in achieving a cure for HIV-1 is the existence of reservoirs of persistent viral infection, with resting memory CD4+ T cells being the largest and best studied. In these cells, HIV-1 rests as a latent provirus refractory to antiretroviral therapy (ART) and invisible to immune surveillance, which can be reawakened and spread the infection. In recent years there has been a push to develop therapies targeting latently infected CD4+ T cells. The "shock and kill" approach seeks to flush latent provirus with "anti-latency" drugs that do not cause global T cell activation, and in the presence of ART to prevent the flushed virus from spreading the infection. However, recent evidence suggests that this approach may be inadequate. Our long-term goal is to develop strategies to directly target and deplete latently infected CD4+ T cells without reactivating the provirus. Our central hypothesis is that surface markers differentially expressed on latently infected CD4+ T cells may be used to achieve that goal. With the support of an NIH-funded grant, we profiled the transcriptome of latently infected and uninfected CD4+ T cells generated with our in vitro latency model, and we identified 33 differentially expressed surface markers. We also found that one of the receptors identified in the microarray screen - CD2 - is expressed at higher levels on latently infected resting memory CD4+ T cells of HIV-1 subjects under suppressive ART. The present application proposes to further pursue these studies with the aim of establishing a panel of cell surface markers that when used in combination will allow to identify with high specificity latently infected resting memory CD4+ T cells in vivo. Thus, in Specific Aim 1 we propose to use our in vitro latency model to validate all the surface markers differentially expressed in the microarray study, and to understand why latently infected cells present that specific expression profile. Specific Aim 2 proposes to validate the markers identified in the previous specific aim, and to generate a panel of surface receptors to define with high specificity latently infected CD4+ T cells in peripheral blood and lymph nodes of HIV-1 patients. The expected outcome of these studies is the establishment of a panel of cell surface markers that in combination will define with high specificity latently infected resting memory CD4+ T cells in vivo. These markers can be used to monitor the size of the latent reservoir in the clinical setting, to enrich latently infcted CD4+ T cells in the laboratory setting, and possibly as a therapeutic target to deplete the latent reservoir without reactivating the provirus. This work is a partnership with Nicolas Chomont at VGTI-Florida, who developed ultra-sensitive techniques to measure integrated HIV-1 provirus in clinical samples, and viral RNA after ex vivo reactivation.

描述（由申请人提供）：实现治愈HIV-1的主要障碍是存在持续性病毒感染库，静止记忆CD4+ T细胞是最大和研究最好的。在这些细胞中，HIV-1作为一种对抗逆转录病毒治疗（ART）难治的潜伏前病毒，对免疫监视不可见，它可以被重新唤醒并传播感染。近年来，人们一直致力于开发针对潜伏感染的CD4+ T细胞的治疗方法。“休克和杀伤”方法寻求用不会引起全局T细胞激活的“抗潜伏期”药物冲洗潜伏的原病毒，并在抗逆转录病毒疗法存在的情况下防止被冲洗的病毒传播感染。然而，最近的证据表明，这种方法可能是不够的。我们的长期目标是开发直接靶向和消耗潜伏感染的CD4+ T细胞而不重新激活原病毒的策略。我们的中心假设是，在潜伏感染的CD4+ T细胞上差异表达的表面标记物可能用于实现这一目标。在美国国立卫生研究院的资助下，我们分析了用体外潜伏期模型生成的潜伏感染和未感染的CD4+ T细胞的转录组，并鉴定了33个差异表达的表面标记。我们还发现，在微阵列筛选中鉴定的受体之一CD2在潜伏上表达的水平更高