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

项目总结/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，或者不能 解决炎症。以前的研究集中在细胞因子和代谢的扰动也表明， ME/CFS患者的CD 8 T细胞反应降低。在初步研究中，我们检查了频率，功能 和CD 8 T细胞的表型状态，以确定它们是否在慢性ME/CFS供体中改变， 与健康的捐赠者相比。我们观察到CD 4：CD 8比率增加， CD 8 T细胞上的CTLA 4和2B 4等耗竭/活化标志物，以及IFN γ、TNF γ和IFN γ的产生减少。 和PMA刺激后CD 107 a/B上调，均提示CD 8 T细胞耗竭。这是 可能与ME/CFS中活化的CD 4 + CD 8 + T细胞的代偿性增加频率相关 与健康对照组相比。值得注意的是，CD 8和CD 4 + CD 8 + T细胞群的亚群 自发产生非典型细胞因子，将ME/CFS供体细分为两个亚群：1型具有非典型细胞因子， 产生IL 9的FoxP 3 +helios+ Treg样细胞的频率增加（女性供体）; 2型具有FoxP 3 +helios- 产生IL 17的细胞（男性供体）。当我们检测CD 4 + CD 8 + T细胞的T细胞受体（TCR）库时， 细胞群我们发现了抗原驱动的克隆扩增到未知抗原的证据， 是病毒抗原还是自身抗原我们假设ME/CFS的共同主题是一种异常的 对感染等免疫触发因素的反应，导致免疫系统永久失调 这是由于CD 8 T细胞耗尽。这些研究将确定潜在的生物标志物和机制， ME/CFS的免疫发病机制导致未来的治疗。我们将在下文中探讨这一假设 目标。目的1：我们将检查ME/CFS中改变的CD 8和CD 4 + CD 8 + T细胞应答：1）我们将确定 CD 8 T细胞耗竭的水平随ME/CFS 1型（女性）和2型（男性）应答而变化， 使用更大的ME/CFS队列研究ME/CFS症状的严重程度; 2）我们将检查EBV抗原特异性应答 在ME/CFS供体中，以确定免疫抑制剂是否改变了常见的持续病毒应答， CD 8 T细胞耗竭的状态并进一步促成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，以帮助理解细胞功能的改变。 在ME/CFS供体中耗尽和活化的CD 8和CD 4 + CD 8 + T细胞亚群。在目标2中，我们将检查TCR CD 8和CD 4 + CD 8 + T细胞亚群的库用于抗原驱动的克隆扩增的证据。定义 活化的克隆扩增的CD 8和CD 4 + CD 8 + T细胞的特性将是该领域的主要步骤 潜在地导致鉴定可能是主要驱动力的特异性感染或自身抗原应答 CD 8 T细胞耗竭的机制和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.