Characterization of Pathogenic T cells in Alopecia Areata

描述(由申请人提供)：为了改进自身免疫患者的治疗方案，更好地确定所涉及的致病T细胞群是至关重要的。为了实现这一点，我们将利用一种新的应用程序，这将首次允许高度集中地剖析致病T细胞库。这一分析将以斑秃为典型例子，弥合迄今在理解自身反应性免疫反应的效应器机制方面存在的差距。由于严重的斑秃病例很少使用全身药物治疗，这种器官特异性自身免疫性疾病代表着一个独特的机会，可以在没有混淆的免疫抑制药物的情况下纵向描述体内自身反应性免疫反应的演变。到目前为止，还没有研究详细跟踪从患者最初的表现到慢性自身免疫的建立的自身反应谱系的详细情况。正在测试的特定假设是，通过结合CDR-3长度谱系分析、磁性分选和流式细胞术，可以在斑秃患者中识别、表征和纵向跟踪致病的自我定向T细胞。目标将是确定：这种致病T细胞反应如何随着时间的推移而演变，致病T细胞的表面表型，以及单个致病T细胞在体内或直接在体外的细胞因子分泌情况。具体目标如下。具体目的1：鉴定斑秃患者活化的克隆型T细胞。具体目标2：确定随着患者疾病的发展，是否纵向存在相同的克隆型T细胞扩增。特异性目标3：检测致病T细胞表面活化标志物、趋化因子受体、整合素黏附分子的表达，以及致病T细胞细胞因子的分泌情况。 公共卫生相关性：在正常情况下，免疫系统可抵御外来入侵病原体，如细菌和病毒。T细胞是免疫系统的一种细胞。这些细胞能够将病毒和细菌的蛋白质与内源性的“自身”蛋白质和构成你身体的细胞区分开来。斑秃是一种自身免疫性疾病。在自身免疫中，通常具有保护性的T细胞错误地将内源性“自身”蛋白识别为外源蛋白，从而导致组织和器官损伤。我们建议研究这些T细胞，以确定自身免疫性疾病的反应是如何随着时间的推移而演变的。我们还将描述自身反应性T细胞的表面及其分泌的分子。

DESCRIPTION (provided by applicant): In order to improve the treatment regimen of patients with autoimmunity it is critical to better define the pathogenic T cell population involved. To accomplish this we will take advantage of a novel application that for the first time will allow a highly focused dissection of the pathogenic T cell repertoire. This analysis will bridge the gap that hitherto existed in understanding the effector mechanisms of an autoreactive immune response, using alopecia areata as a prototypic example. Since severe cases of alopecia areata are rarely managed with systemic agents, this organ-specific autoimmune disease represents a unique opportunity to longitudinally characterize the evolution of an autoreactive immune response in vivo in the absence of confounding immunosuppressive medications. No study to date has followed in detail an autoreactive repertoire longitudinally from a patient's initial presentation to the establishment of chronic autoimmunity. The specific hypothesis being tested is that pathogenic self-directed T cells can be identified, characterized and followed longitudinally in patients with alopecia areata by combining CDR-3 length repertoire analysis with magnetic sorting and flow cytometry. The goal will be to determine: how this pathogenic T cell response evolves over time, the surface phenotype of the pathogenic T cells, and the in vivo or directly ex vivo cytokine secretion profile of the individual pathogenic T cells. The specific aims are as follows. Specific Aim 1: Identify activated clonotypic T cells in patients with alopecia areata. Specific Aim 2: Determine if the same clonotypic T cell expansions are present longitudinally as a patient's disease evolves. Specific Aim 3: Determine the pathogenic T cell's surface expression of activation markers, chemokine receptors, and integrin adhesion molecules as well as the pathogenic T cell's cytokine secretion profile. PUBLIC HEALTH RELEVANCE: Under normal circumstances the immune system protects against foreign invading pathogens such as bacteria and viruses. The T cells are a type of cell of the immune system. These cells have the ability to distinguish viral and bacterial proteins from enogenous "self" proteins and cells that make up your body. Alopecia areata is an autoimmune disease. In autoimmunity the usually protective T cells mistakingly recognize enogenous "self" proteins as foreign, which leads to tissue and organ damage. We propose to study these T cells to determine how the autoimmune disease response evolves over time. We will also characterize the surface of the autoreactive T cells and the molecules they secrete.