Gamma delta T cells promote inflammation in aviremic HIV infection and normal aging

摘要 即使成功地控制了病毒，艾滋病毒感染者也表现出与老年相关的共病， 包括骨质疏松症、中风、痴呆症和癌症。在艾滋病病毒携带者和普通老年人中 人口、年龄相关疾病和死亡率与血浆炎症和肠道标志物相关 渗透性。肠道是潜伏感染艾滋病毒的细胞的主要储存库，以及艾滋病毒肠病，定义为 小肠和结肠的病理过程是HIV感染的一个标志。我们的初步数据 在ART抑制的艾滋病毒感染和正常的情况下，牵涉到T细胞作为炎症驱动因素 衰老。T细胞是一种非传统的T细胞谱系，占循环T细胞的10%，但在≤中发现 肠道上皮细胞的比例要高得多。此外，有证据表明，这种独特的T细胞 在正常情况下，人群调节肠道屏障功能，T促炎活性 对上皮部位和上皮屏障造成损害。因此，我们假设与病毒有关 抑制艾滋病毒感染和正常衰老的胃肠道T细胞通过直接窝藏 HIV和/或暴露于炎症因子，这种异常激活导致紧密连接的破坏 肠道上皮屏障，导致微生物产物和炎性T细胞释放增加 发行量。此外，我们预测老化的T细胞表现出偏向炎症性的功能图谱 细胞因子/细胞毒性对艾滋病毒直接感染和/或刺激因素的反应。在此应用程序中，我们 建议使用先进和创新的方法来测试这些假设，包括25色流式细胞术， 直肠乙状结肠活检的31色成像质量细胞术、血浆和细胞培养的19和33复合体分析 上清液，以及用于收集数据集的多变量分析的多种算法。我们的 生物信息学数据分析计划将能够识别新的T细胞亚群并分析差异影响 感染和不感染艾滋病毒的年龄。在目标1中，我们将对我们的艾滋病毒和老龄化进行横断面研究 确定循环T细胞亚群、血浆炎症和肠道之间联系的队列研究 通透性标记物、肠道结构和细胞组成。在目标2中，我们将确定 T细胞亚群、血浆标志物与老年性痴呆的发病和/或进展之间的时间联系 通过对患有和不患有抗逆转录病毒药物抑制的艾滋病毒的老年受试者进行纵向研究的结果。在《目标3》中，我们将 进行体外试验，以确定年龄和艾滋病毒感染如何影响T细胞功能，包括能力 分解肠上皮细胞单层。我们预测，我们提议的实验将确定 导致全身性炎症和年龄相关性并存增加的生物学机制 艾滋病病毒携带者和一般老年人群；这样的洞察力可能导致 减少“炎症老化”相关疾病和死亡的新疗法。

ABSTRACT Even with successful viral control, HIV-infected individuals exhibit co-morbidities associated with older age, including osteoporosis, stroke, dementia, and cancer. In aviremic HIV+ individuals and the general geriatric population, age-associated diseases and mortality correlate with plasma markers of inflammation and intestinal permeability. The gut is a major reservoir of latently HIV-infected cells, and HIV enteropathy, defined as pathologic processes in the small intestine and colon, is a hallmark of HIV infection. Our preliminary data implicate gamma delta ( T cells as an inflammatory driver in ART-suppressed HIV infection and with normal aging.  T cells are a non-conventional T cell lineage that comprise ≤10% of circulating T cells yet are found in considerably higher proportions in the epithelium of the intestine. Also, there is evidence that this unique T cell population regulates intestinal barrier function during normal conditions, and that  T pro-inflammatory activity causes damage at epithelial sites and to epithelial barriers. Therefore, we hypothesize that with virally suppressed HIV infection and with normal aging, gastrointestinal T cells are stimulated via directly harboring HIV and/or exposure to inflammatory factors and this aberrant activation leads to breakdown of tight junctions of the intestinal epithelial barrier, causing increased release of microbial products and inflammatory  T cells into the circulation. Further, we predict that aged  T cells exhibit functional profiles skewed towards inflammatory cytokines/cytotoxicity in response to either direct HIV infection and/or stimulatory factors. In this application, we propose to test these hypotheses using advanced and innovative approaches, including 25-color flow cytometry, 31-color imaging mass cytometry of recto-sigmoid biopsies, 19- and 33-plex analyses of plasma and cell culture supernatants, respectively, and multiple algorithms for multivariate analysis of collected datasets. Our bioinformatic data analysis plan will enable identifying novel  T cell subsets and parsing the differential impacts of age with and without HIV infection. In Aim 1 we will perform a cross-sectional study of our HIV and Aging cohort to determine the links between circulating  T cell subsets, plasma inflammatory and intestinal permeability markers, and intestinal architecture and cellular composition. In Aim 2 we will determine the temporal links between  T cell subsets, plasma markers, and the onset and/or progression of geriatric outcomes via a longitudinal study of older subjects with and without ART-suppressed HIV. In Aim 3, we will perform in vitro assays to determine how age and HIV infection impact  T cell functions, including the capacity to breakdown intestinal epithelial cell monolayers. We predict that our proposed experiments will identify the biological mechanisms that drive the increased systemic inflammation and age-associated comorbidities in both aviremic HIV+ individuals and the general geriatric population; such insight could lead to the development of novel therapeutics to reduce ‘inflamm-aging’-associated diseases and deaths.