Mechanisms of CD8+ T cell Immune surveillance in human genital skin and mucosa af

CD8 T细胞对人类生殖器皮肤和粘膜免疫监测的机制

基本信息

批准号：
8508374
负责人：
Jia Zhu
金额：
$ 31.75万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8508374
关键词：
Affect Anatomic Sites Antiviral Agents Binding Biopsy Specimen Blood Blood Circulation CD8-Positive T-Lymphocytes CD8B1 gene Cell Cycle Progression Cells Clinical Clonality Containment Cytolysis Dermal Development Disease Evolution Gene Expression Genes Genital system Genome HIV Healed Hour Human Human Herpesvirus 2 Immune Immune response Immune system Immunity Immunologic Monitoring Immunologic Surveillance Immunotherapeutic agent In Situ Infection Kinetics Knowledge Lesion Mediating Metabolic Metabolic Activation Methods Molecular Molecular Profiling Mucous Membrane Normal tissue morphology Outcome Pathogenesis Patients Peripheral Physiological Population Property Proteins Risk Factors Sensory Nerve Endings Simplexvirus Site Skin Symptoms T memory cell T-Cell Activation T-Cell Receptor T-Lymphocyte Technology Tissues Ulcer Vaccines Viral Viral Antigens Virion Virus Work cell type clinical phenotype cohort design healing in vivo laser capture microdissection lymphocyte proliferation new technology novel pandemic disease particle pathogen prevent prophylactic public health priorities self-renewal success transmission process

项目摘要

DESCRIPTION (provided by applicant): The human pathogen herpes simplex virus type 2 (HSV-2) is a leading cause of genital ulcer disease and a risk factor for human immunodeficiency virus (HIV) acquisition and transmission. Developing an effective HSV-2 vaccine is a public health priority for controlling the pandemic; however success has been limited largely due to our poor understanding of protective immunity required for containing HSV-2 infection and reactivation in humans. Our work has shown that the peripheral mucosal immune system, specifically HSV specific CD8+ T cells, persist in the periphery at the dermal-epidermal junction (DEJ) contiguous to the sensory nerve endings where HSV virion particles are released, and that HSV-2 specific CD8+ T cells frequently encounter viral antigens. Recent studies have shown that over 80% of HSV-2 reactivations are subclinical and of short duration, typically lasting only 6 to 12 hours, indicating that a rapid host immune response in the genital skin and mucosa can contain local HSV reactivation thus preventing symptoms. We hypothesize that persistent CD8+ T cells at the dermal-epidermal junction exert antiviral effector functions upon re-encountering viral antigen and rapidly eliminate infected cells before the virus can cause clinically symptomatic disease. We have recently developed novel technologies to define, in situ, the mechanisms by which HSV specific CD8+ T cells contain virus in the periphery, a key concept for the rational design and development of novel immunotherapeutic as well as prophylactic vaccines against herpes simplex virus. We have developed and perfected a cell-type specific laser capture microdissection (LCM) method. Using genome transcriptional profiles of LCM-CD8+ T cells, we have shown that CD8+ T cells at the dermal-epidermal junction (DEJ CD8+) have expression profiles consistent with cytolytic activity, metabolic activation and proliferative potential. We have also applied T cell receptor (TCR) high-throughput sequencing technology to LCM-CD8+ cells to determine the breadth and clonality of the CD8 TCR repertoire in locally affected skin/mucosa tissue and in blood. These studies allow us to compare T cell populations at the dermal-epidermal junction and dermal perivascular sites, and to evaluate, at the TCR sequence level, their T cell clonotypes and dominance over the course of herpes lesion evolution. The unique ability of our group to obtain sequential biopsy samples of HSV infected genital lesions and normal tissue, and to study patient cohorts with distinct clinical phenotypes, will yield opportunities to not only increase knowledge about HSV pathogenesis, but also the kinetics and mechanistic functions of resident mucosal T cells in HSV-2 containment. This proposal is designed to define immune control mechanisms and proliferation potentials for CD8+ T cells at their original anatomic sites and native physiological state in skin and mucosa, thus characterizing the molecular interactions between HSV-2 and human host in vivo.

描述（由申请人提供）：人类病原体单纯疱疹2型（HSV-2）是生殖器溃疡疾病的主要原因，也是人类免疫缺陷病毒（HIV）获取和传播的危险因素。开发有效的HSV-2疫苗是控制大流行的公共卫生优先事项。但是，成功的限制很大程度上是由于我们对含有HSV-2感染和人类重新激活所需的保护性免疫的不良理解。我们的工作表明，外围粘膜免疫系统，特别是HSV特异性CD8+ T细胞，在皮肤 - 表皮连接（DEJ）的外围持续存在于与释放HSV Virion颗粒的感觉神经端的连续，并且HSV-2特异性CD8+ T型细胞经常经常遇到病毒抗原剂。最近的研究表明，超过80％的HSV-2重新激活是亚临床的，持续时间短，通常仅持续6至12个小时，表明生殖器皮肤和粘膜中快速宿主免疫反应可以包含局部HSV重新激活，从而防止症状。我们假设皮肤表皮连接处的持久性CD8+ T细胞在重新遇到病毒抗原后会发挥抗病毒效应子功能，并在病毒引起临床症状性疾病之前迅速消除受感染的细胞。我们最近开发了新型技术来原位定义HSV特异性CD8+ T细胞在外围含有病毒的机制，这是新型免疫治疗性以及针对疱疹病毒的预防性疫苗的合理设计和开发的关键概念。我们已经开发并完善了细胞类型的特异性激光捕获微分解（LCM）方法。使用LCM-CD8+ T细胞的基因组转录谱，我们已经表明，皮肤表皮连接处的CD8+ T细胞（DEJ CD8+）具有与细胞溶解活性，代谢激活和增殖潜力一致的表达谱。我们还将T细胞受体（TCR）高通量测序技术应用于LCM-CD8+细胞，以确定局部受影响的皮肤/粘膜组织和血液中CD8 TCR库的广度和克隆性。这些研究使我们能够比较皮肤 - 表皮结和皮肤血管周围部位的T细胞群体，并在TCR序列水平上评估其T细胞clonotypes和在疱疹病变进化过程中的优势。我们小组获得HSV感染的生殖器病变和正常组织的顺序活检样本的独特能力，以及研究具有不同临床表型的患者同类群，将产生机会，不仅会增加有关HSV-2容器中HSV-2细胞的动力学和机械性功能的知识，还可以增加有关HSV-2容器中HSV-2细胞的动力学的知识。该建议旨在定义CD8+ T细胞的免疫控制机制和原始解剖部位和天然生理学的增殖潜力皮肤和粘膜中的状态，从而表征了HSV-2与体内人类宿主之间的分子相互作用。