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

项目概要/摘要 肌痛性脑脊髓炎/慢性疲劳综合征（ME/CFS）是一种影响众多器官的复杂疾病 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，或无法 解决炎症。先前针对细胞因子和代谢扰动的研究也表明 ME/CFS 中 CD8 T 反应降低。在初步研究中，我们检查了频率、功能 和 CD8 T 细胞的表型状态，以确定它们在慢性 ME/CFS 供体中是否发生改变 与健康捐献者相比。我们观察到 CD4:CD8 比率增加， CD8 T 细胞上的耗竭/激活标记物如 CTLA4 和 2B4，以及 IFNα、TNFα 的产生减少 PMA 刺激后 CD107a/b 上调，所有这些都表明 CD8 T 细胞耗竭。这是 与 ME/CFS 中激活的 CD4 CD8 T 细胞的频率可能补偿性增加相关 捐赠者与健康对照者相比。值得注意的是，CD8 和 CD4 CD8 T 细胞群的一个子集 自发产生非典型细胞因子，将 ME/CFS 供体细分为两个子集：1 型具有 FoxP3 helios Treg 样细胞产生 IL9 的频率增加（女性供体）； 2型有FoxP3 helios- 产生 IL17 的细胞（男性供体）。当我们检查 CD4 CD8 T 的 T 细胞受体 (TCR) 库时 我们此时在细胞群中发现了抗原驱动的克隆扩展到未知抗原的证据， 它是病毒抗原还是自身抗原。我们假设 ME/CFS 的共同主题是异常的 对感染等免疫触发因素的反应，导致免疫系统永久性失调 CD8 T 细胞耗竭的结果。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了未来的治疗。我们将在下面探讨这个假设 目标。目标 1 我们将检查 ME/CFS 中 CD8 和 CD4 CD8 T 细胞反应的改变：1) 我们将确定是否 CD8 T 细胞耗竭水平随 ME/CFS 1 型（女性）和 2 型（男性）反应以及 使用更大的 ME/CFS 队列来确定 ME/CFS 症状的严重程度； 2) 我们将检查 EBV 抗原特异性反应 在 ME/CFS 供体中进行研究，以确定免疫抑制剂是否会改变常见的持续病毒反应 CD8 T 细胞耗竭状态，进一步导致 ME/CFS 的疾病状态； 3) 微阵列分析 将在分选的活化 T 细胞亚群上进行，以帮助理解 T 细胞功能的变化 ME/CFS 供体中 CD8 和 CD4 CD8 T 细胞亚群耗尽并激活。在目标 2 中，我们将检查 TCR CD8 和 CD4 CD8 T 细胞亚群的库，用于抗原驱动克隆扩张的证据。定义 激活的克隆扩增 CD8 和 CD4 CD8 T 细胞的特征将是该领域的重要一步 可能导致识别出可能是主要驱动因素的特定感染性或自身抗原反应 CD8 T 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.