Altered T Cell Responses in Myalgic Enchephalomeylitis/Chronic Fatigue Syndrome (ME/CFS)

项目摘要/摘要 肌痛性脑脊髓炎/慢性疲劳综合征(ME/CFS)是一种累及多个器官的复杂疾病 系统和生物过程。已发表的数据似乎表明，ME/CFS可能先于感染， 疾病的慢性表现可能代表宿主对感染的反应改变，或者无法 消炎消炎。以前的研究集中在细胞因子和新陈代谢的扰动上，也表明 ME/CFS患者CD8T反应降低。在初步研究中，我们检查了频率、功能 和CD8T细胞的表型状态，以确定其在慢性ME/CFS供者中是否发生了改变 与健康的捐赠者相比。我们观察到CD4：CD8比率增加，表达改变 CD8T细胞表面CTLA4和2B4等耗竭/激活标志物以及干扰素、肿瘤坏死因子的产生减少 PMA刺激后CD107a/b表达上调，均提示CD8T细胞耗竭。这是 与ME/CFS中活化的CD8T细胞的频率可能代偿性增加有关 捐献者与健康对照相比。值得注意的是，CD8和CD4 CD8 T细胞亚群 自发产生非典型细胞因子，将ME/CFS捐赠者细分为两个子集：类型1具有 FoxP3 Helios Treg样细胞产生IL9的频率增加；2型有FoxP3 Helios- 产生IL17(男性供体)的细胞。当我们检测CD4CD8T的T细胞受体(TCR)谱系时 细胞群我们发现了抗原驱动的克隆扩张到未知抗原的证据， 无论它是病毒还是自身抗原。我们假设ME/CFS的共同主题是反常的 对免疫触发的反应，如感染，导致永久性免疫系统失调 由于CD8 T细胞耗尽所致。这些研究将确定潜在的生物标志物和驱动机制 ME/CFS的免疫发病机制导致未来的治疗。我们将在以下内容中探讨这一假设 目标。目的1)我们将检测ME/CFs中改变的CD8和CD4CD8 T细胞反应：1)我们将确定 CD8 T细胞耗竭水平随ME/CFS 1型(女性)和2型(男性)的反应以及 使用更大的ME/CFS队列研究ME/CFS症状的严重性；2)我们将检查EBV抗原特异性反应 在ME/CFS供体中确定免疫抑制是否改变了常见的持久性病毒反应 CD8 T细胞耗竭状态及其与ME/CFS疾病状态的关系；3)基因芯片分析 将在已排序的激活T细胞亚群上完成，以帮助了解 ME/CFS供者的CD8和CD4CD8 T细胞亚群的耗竭和激活。在目标2中，我们将研究TCR CD8和CD4 CD8 T细胞亚群，以寻找抗原驱动的克隆性增殖的证据。定义 活化的克隆扩增的CD8和CD4 CD8 T细胞的特性将是该领域的重要一步 可能导致确定一种可能是主要驱动力的特定的感染性或自身抗原反应 CD8T细胞耗竭和ME/CFS的免疫学基础。

Project Summary/Abstract Myalgic enchephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex disorder affecting numerous organ systems and biological processes. Published data seems to suggest that ME/CFS may be preceded by infection, and the chronic manifestation of illness may represent an altered host response to infection, or an inability to resolve inflammation. Previous studies focused on perturbation in cytokines and metabolism have also shown that CD8 T responses are decreased in ME/CFS. In preliminary studies we examined the frequency, functional and phenotypic status of CD8 T cells to determine whether they were altered in chronic ME/CFS donors as compared to healthy donors. We observed an increased CD4:CD8 ratio, altered expression of exhaustion/activation markers like CTLA4 and 2B4 on CD8 T cells, and decreased production of IFN, TNF and CD107a/b upregulation following PMA stimulation, all suggesting CD8 T cell exhaustion. This was associated with a, perhaps compensatory increased frequency of activated CD4+CD8+ T cells in the ME/CFS donors as compared to healthy controls. Notably, a subset of the CD8 and the CD4+CD8+ T cell populations were spontaneously producing atypical cytokines, subdividing ME/CFS donors into two subsets: type 1 had an increased frequency of FoxP3+helios+ Treg-like cells producing IL9 (female donors); type 2 had FoxP3+helios- cells producing IL17 (male donors). When we examined the T-cell receptor (TCR) repertoire of the CD4+CD8+ T cell population we found evidence of antigen driven clonal expansions to an unknown antigen at this time, whether it will be a viral or auto-antigen. We hypothesize that the common theme in ME/CFS is an aberrant response to an immunological trigger like infection, which results in a permanently dysregulated immune system as a result of CD8 T cell exhaustion. These studies will identify potential biomarkers and mechanisms driving the immunopathogenesis of ME/CFS leading to future therapies. We will explore this hypothesis in the following Aims. Aim 1 we will examine altered CD8 and CD4+CD8+ T-cell responses in ME/CFS: 1) we will determine if the level of CD8 T cell exhaustion varies with ME/CFS type 1 (female) and Type 2 (male) response and with the severity of ME/CFS symptoms using a larger ME/CFS cohort; 2) we will examine EBV antigen-specific responses in ME/CFS donors to determine if a common persistent virus response is altered by the immunosuppressive state of CD8 T cell exhaustion and further contributing to the disease state of ME/CFS; 3) microarray analyses will be done on sorted activated T cell subsets to assist in understanding the alterations in the functionality of the exhausted and activated CD8 and CD4+CD8+ T cell subsets in ME/CFS donors. In Aim 2 we will examine TCR repertoire of CD8 and CD4+CD8+ T-cell subsets for evidence of antigen driven clonal expansion. Defining the characteristics of the activated clonally expanded CD8 and CD4+CD8+ T cells would be a major step in the field potentially leading to the identification a specific infectious or auto-antigen response that could be the main driver of CD8 T cell exhaustion and the immunological basis of ME/CFS.