Autoreactive T Cell Function in Vitiligo

白癜风中的自身反应性 T 细胞功能

• 批准号: 10404445
• 负责人: SALLY Choate KENT
• 金额: $ 50.69万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404445
• 关键词: 3-Dimensional; Affect; Affinity; Animal Model; Antigen Targeting; Antigenic Specificity; Antigens; Autoimmune Diseases; Autoimmunity; Biological Assay; Blood; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Communication; Cell Culture Techniques; Cell physiology; Cells; Characteristics; Clinical; Clonality; Clone Cells; Communication; Complex; Computing Methodologies; Data; Disease; Effector Cell; Epidermis; Epigenetic Process; Fluorescent in Situ Hybridization; Gene Expression Profile; Genetic; Genetic Risk; Human; Immune; Immunohistochemistry; In Vitro; Individual; Inflammatory; Insulin-Dependent Diabetes Mellitus; Lesion; Location; Maintenance; Mediating; Microtome - medical device; Modeling; Molecular; Multiomic Data; Multiple Sclerosis; Organ; Organ Culture Techniques; Pathway interactions; Patients; Phenotype; Pigments; Play; Population; Positioning Attribute; Proteins; Regulatory T-Lymphocyte; Resolution; Role; Salvelinus; Sampling; Signal Transduction; Skin; Slice; Stable Disease; T cell clonality; T cell receptor repertoire sequencing; T cell response; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic Intervention; Tissues; Transcript; Translational Research; Trichrome stain method; Vitiligo; White Spots; autoreactive T cell; base; clinical phenotype; cytokine; cytotoxic; cytotoxic CD8 T cells; disease phenotype; effector T cell; health disparity; human tissue; in vivo; melanocyte; mouse model; multiple omics; novel therapeutic intervention; programs; recruit; response; risk variant; single-cell RNA sequencing; skin disorder; skin lesion; spatial relationship; tool

T cells are effector cells of autoimmunity that migrate to find their targets, produce cytokines that recruit and activate other immune cells, and destroy their target cells in affected tissue. Vitiligo is an autoimmune disease of the skin in which cytotoxic CD8+ T cells target pigment-making melanocytes, which results in disfiguring white spots that are particularly devastating for those with darker skin and thus leads to health disparities for the most vulnerable of our population. Vitiligo is an ideal disease in which to investigate mechanisms of organ- specific autoimmunity because disease phenotype can be directly correlated to molecular pathways. That is, affected skin can be observed and sampled, target cells and antigens are known, and translational research tools are available. Vitiligo shares genetic risk alleles and other mechanisms with autoimmune diseases like type 1 diabetes and multiple sclerosis that are more difficult to study in human patients. Through vitiligo we can develop a comprehensive understanding of organ-specific autoimmunity as it progresses within human tissue. In vitiligo and models of other human autoimmune diseases, autoreactive T cell responses are commonly char- acterized as homogenous, with an “all or nothing” result. This is partly due to the fact that mouse models of autoimmunity are frequently based on a single high-affinity CD8+ cytotoxic clone transferred and activated arti- ficially. However, we found that T cells infiltrating vitiligo skin lesions are polyclonal and heterogeneous, with different antigenic specificities and activation states. Cytotoxic CD8+ T cells organize into clustered units within the epidermis that appear to attack melanocytes, while CD4+ and Treg cells are at the periphery of these clus- ters. Together, these data indicate that T cell interactions in vitiligo are more complex than previously under- stood. Our central hypothesis is that polyclonal CD8+, CD4+, and Treg cell communications coordinate mela- nocyte destruction and determine vitiligo clinical phenotypes. This project will determine how autoreactive T cell clonal diversity and localization define the clinical disease phenotype, how T cell subtypes interact with each other to coordinate autoimmunity, and how T cells orches- trate melanocyte destruction within the skin. First, we will determine the in vivo location, spatial relationships, and functional characteristics of T cell clones in different clinical phenotypes of vitiligo, such as early incipient vs. late established lesions, active vs. stable disease, and single vs. multiple locations. Next, we will character- ize the function and reactivity of T cells isolated directly from the skin of these lesions. We will use primary T cell and skin organ culture, T cell functional assays, T cell receptor affinity assays, the sequential fluorescence in situ hybridization (seqFISH+) Core, and advanced computational methods. This project is translational in approach because it will create new mechanistic understanding of how T cells mediate the initiation, progres- sion, and maintenance of autoimmunity during vitiligo. Together these aims will define molecular mechanisms utilized by autoreactive T cells and identify key functional pathways, supporting new therapeutic interventions.

T 细胞是自身免疫的效应细胞，它们会迁移以寻找目标，产生细胞因子来招募和 激活其他免疫细胞，并破坏受影响组织中的目标细胞。白癜风是一种自身免疫性疾病 细胞毒性 CD8+ T 细胞靶向产生色素的黑素细胞，从而导致毁容的皮肤 白斑对于肤色较深的人来说尤其具有破坏性，从而导致健康差异 我们人口中最弱势的群体。白癜风是研究器官机制的理想疾病 特异性自身免疫，因为疾病表型可以与分子途径直接相关。那是， 受影响的皮肤可以被观察和取样，靶细胞和抗原是已知的，并且转化研究 工具可用。白癜风与自身免疫性疾病具有相同的遗传风险等位基因和其他机制，例如 1 型糖尿病和多发性硬化症在人类患者中的研究更加困难。通过白癜风我们可以 全面了解器官特异性自身免疫在人体组织内的进展。 在白癜风和其他人类自身免疫性疾病模型中，自身反应性 T 细胞反应通常具有以下特点： 其特征是同质的，具有“全有或全无”的结果。这部分是由于以下事实：小鼠模型 自身免疫通常基于转移和激活的单个高亲和力 CD8+ 细胞毒性克隆 正式地。然而，我们发现浸润白癜风皮损的T细胞是多克隆且异质的， 不同的抗原特异性和激活状态。细胞毒性 CD8+ T 细胞在内部组织成簇状单元 表皮似乎攻击黑色素细胞，而 CD4+ 和 Treg 细胞位于这些细胞的外围 之三。总之，这些数据表明白癜风中的 T 细胞相互作用比以前所认为的更加复杂。 站着。我们的中心假设是多克隆 CD8+、CD4+ 和 Treg 细胞通讯协调 mela- 细胞破坏并确定白癜风临床表型。 该项目将确定自身反应性 T 细胞克隆多样性和定位如何定义临床疾病 表型、T 细胞亚型如何相互作用以协调自身免疫，以及 T 细胞如何协调- 治疗皮肤内黑色素细胞的破坏。首先，我们要确定体内位置、空间关系、 白癜风不同临床表型（如早期）中T细胞克隆的功能特征 与晚期形成的病变、活动性与稳定疾病、以及单个位置与多个位置。接下来我们将角色—— 确定直接从这些病变皮肤中分离出的 T 细胞的功能和反应性。我们将使用初级 T 细胞和皮肤器官培养、T 细胞功能测定、T 细胞受体亲和力测定、序贯荧光 原位杂交 (seqFISH+) 核心和先进的计算方法。该项目已翻译为 方法，因为它将对 T 细胞如何介导启动、进展产生新的机制理解。 白癜风期间维持自身免疫。这些目标将共同定义分子机制 被自身反应性 T 细胞利用并确定关键功能途径，支持新的治疗干预措施。