Epigenetic Control of CD8 T-Cell Differentiation in Human Lymph Nodes

人类淋巴结 CD8 T 细胞分化的表观遗传控制

基本信息

批准号：
9767954
负责人：
Zaza Mtine Ndhlovu
金额：
$ 14.47万
依托单位：
KWAZULU-NATAL RESEARCH INSTITUTE TB-HIV
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-07 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9767954
关键词：
ATAC-seq Accounting Acute Address Adherence Animals Area B-Lymphocytes BLR1 gene Binding Biological Assay CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CD8B1 gene Caring Cells Chromatin Chronic Chronic Disease DNA Data Detection Development Education Epidemic Epigenetic Process Exclusion Failure Female Flow Cytometry Foundations Frequencies Future Gene Expression Gene Expression Profiling Genes Genetic Transcription HIV HIV Infections HIV-1 Health Heart Helper-Inducer T-Lymphocyte High-Throughput Nucleotide Sequencing Home environment Human Image Immune Immune response Immune system Immunotherapeutic agent In Vitro Individual Infection Interleukin-12 Interleukin-15 Interruption Intervention Knowledge Lead Life Logistics Longitudinal cohort Lymph Node Tissue Lymph node excision Lymphoid Tissue MADH4 gene Mediating Methods Molecular Molecular Conformation Monitor Mus Oral Persons Phenotype Plasma Population Problem Solving RNA Regulation Sampling Site Source South Africa Structure of germinal center of lymph node T cell differentiation TCF Transcription Factor TCF3 gene Testing Tissues Transcription Initiation Site Transforming Growth Factor beta Transposase Tumor-infiltrating immune cells Ursidae Family Viral Load result Viremia Virus antiretroviral therapy base cell motility chemokine receptor chronic infection cohort cost cytokine cytotoxic differential expression epigenetic drug epigenome follow-up genome-wide human tissue immune clearance immune function innovation interleukin-23 lymph nodes novel novel strategies pill pre-exposure prophylaxis preservation programs recruit repository transcription factor transcriptome sequencing transmission process treatment program young woman

项目摘要

PROJECT SUMMARY The development of potent antiretroviral therapies, now delivered as a single pill once a day, has transformed HIV infection into a chronic disease. Globally, over 19 million people are now on life-long treatment, and test- and-treat strategies as was well as oral PrEP have the ability to further reduce HIV transmissions. However, despite these remarkable advances, prolonged ART mediated suppression of plasma viral loads to undetectable levels does not eradicate the virus, which rapidly rebounds upon treatment interruption. The many logistical limitations and cost challenges that come with providing life-long care to those living with HIV highlights the need for novel strategies of controlling the virus in the absence of therapy. Emerging data indicate that lymph nodes are a major source of HIV persistence during ART mainly because infected T follicular helper cells are protected from immune elimination due to partial exclusion of cytotoxic CD8 T cells (CTL) from germinal centers (GC). The molecular mechanisms that regulate CXCR5 expression which allows CTL migration into GCs are not known, mainly due to the difficulty associated with obtaining lymph node samples for such studies. Through innovative recruitment strategies, we have now solved this problem. We propose to use stored lymph node samples obtained from persons identified and treated at the onset of plasma viremia, in many when plasma viral loads are less than 1,000 RNA copies/ml to investigate why CTLs are largely excluded from GCs. Our preliminary ATAC-Seq studies suggest that CXCR5 expression on lymph node CTLs is epigenetically regulated. To follow up on this observation we propose to conduct chromatin accessibility analysis and transcriptional profiling of sorted HIV-specific CD8 T cells to identify epigenetic mechanisms that regulate CXCR5 expression. Our experimental approach will involve pairwise chromatin accessibility and transcription analysis of sorted follicular and extrafollicular HIV-specific CD8 T cells. This strategy will help to identify novel epigenetic and transcription factors that are different between the two populations. Finally, we will manipulate some of the differentially expressed genes using exogenous agents namely cytokines and epigenetic modifying drugs to determine the optimal conditions to induce CXCR5 expression on CD8 T cells. Our results will lead to novel strategies for redirecting CTL to follicular areas where they are needed to kill HIV infected cells. If successful, our studies will lead to novel strategies for redirecting CTL to follicular areas to eradicate HIV infection as part of a cure strategy.

项目概要有效的抗逆转录病毒疗法的开发，现在以每天一次的单一药丸形式提供，已经改变了 HIV感染变成一种慢性疾病。在全球范围内，目前有超过 1900 万人正在接受终身治疗，并且正在接受测试- 治疗策略以及口服 PrEP 能够进一步减少 HIV 传播。然而，尽管取得了这些显着的进步，长期 ART 介导的血浆病毒载量抑制至无法检测到水平并不能根除病毒，治疗中断后病毒会迅速反弹。许多后勤向艾滋病毒感染者提供终身护理所带来的限制和成本挑战凸显了这一需求在没有治疗的情况下控制病毒的新策略。新出现的数据表明，淋巴结是 ART 期间 HIV 持续存在的一个主要原因，主要是因为受感染的滤泡辅助 T 细胞受到保护由于细胞毒性 CD8 T 细胞 (CTL) 被部分排除在生发中心 (GC) 之外，从而导致免疫消除。这调节 CXCR5 表达从而允许 CTL 迁移到 GC 的分子机制尚不清楚，主要是由于此类研究难以获取淋巴结样本。通过创新招聘策略，我们现在已经解决了这个问题。我们建议使用储存的淋巴结样本从血浆病毒血症发作时被识别和治疗的人获得，在许多情况下，血浆病毒载量低于 1,000 个 RNA 拷贝/ml，以调查为什么 CTL 很大程度上被排除在 GC 之外。我们的初步 ATAC-Seq 研究表明，淋巴结 CTL 上的 CXCR5 表达受到表观遗传调控。关注根据这一观察，我们建议进行染色质可及性分析和转录谱分析对 HIV 特异性 CD8 T 细胞进行分类，以确定调节 CXCR5 表达的表观遗传机制。我们的实验方法将涉及分选滤泡的成对染色质可及性和转录分析和滤泡外 HIV 特异性 CD8 T 细胞。该策略将有助于识别新的表观遗传和转录两个人群之间存在差异的因素。最后，我们将操纵一些差异使用外源性试剂（即细胞因子和表观遗传修饰药物）表达基因，以确定诱导 CD8 T 细胞上 CXCR5 表达的最佳条件。我们的研究结果将带来新的策略将 CTL 重定向到需要它们杀死 HIV 感染细胞的毛囊区域。如果成功的话，我们的研究将导致将 CTL 重定向到毛囊区域以根除 HIV 感染的新策略，作为治疗策略的一部分。