Determining the mechanisms of vascular CD8 T cell activation in CMV and HIV infections

确定 CMV 和 HIV 感染中血管 CD8 T 细胞激活的机制

基本信息

批准号：
10461964
负责人：
Michael L Freeman
金额：
$ 20.13万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-04 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10461964
关键词：
AIDS/HIV problem Age Animal Model Antibodies Antigens Aorta Arterial Fatty Streak Atherosclerosis Automobile Driving Bar Codes Blood Vessels CD58 gene CD8-Positive T-Lymphocytes CX3CL1 gene Cardiovascular Diseases Cell physiology Cells Chronic Coupled Cytomegalovirus Cytomegalovirus Infections Development Elderly Endothelium Exhibits Fluorescent in Situ Hybridization Foundations Future Gender Genetic Transcription HIV HIV Infections Homing Human Image Immunofluorescence Immunologic Impairment In Situ In Vitro Inflammation Inflammatory Interleukin-15 Knowledge Label Lead Link Mediating Morbidity - disease rate Myeloid Cells Outcome Pathogenicity Pathology Pathway interactions Peptides Persons Phenotype Plague Population Prevalence Prevention strategy RNA Research Research Priority Risk Risk Factors Shapes Signal Transduction Site Smooth Muscle Myocytes Source Specificity Specimen T cell response T-Cell Activation T-Lymphocyte Testing Time Tissues United States National Institutes of Health Vascular Endothelium Viral Physiology Virus Diseases antiretroviral therapy cardiovascular disorder risk co-infection comorbidity endothelial dysfunction immune activation in vivo innovation monocyte novel paracrine pathogen prevent programs receptor single cell analysis single-cell RNA sequencing spatial relationship therapy development transcriptome sequencing treatment strategy

项目摘要

PROJECT SUMMARY/ABSTRACT Persons living with HIV infection (PWH) on antiretroviral therapy (ART) are succumbing earlier to many of the same complications that plague the HIV-uninfected elderly. This elevated CVD risk in PWH is at least partly due to persistently increased inflammation and activation of CD8 T cells and monocytes that our group has identified and characterized. Many elderly and nearly all PWH are coinfected with cytomegalovirus (CMV), which we have shown is associated with inflammation and dramatic expansion of CD8 T cells in PWH, and has been independently linked to CVD. Activated CD8 T cells accumulate at sites of endothelial dysfunction in vivo, promote atherosclerotic CVD (ASCVD) in animal models, and can trigger a proatherogenic phenotype on monocytes in vitro – yet whether CD8 T cells infiltrating the vasculature recognize cognate antigenic peptides has never been definitively shown in vivo, nor have antigen-nonspecific activating pathways been rigorously evaluated. In this proposal, we seek to identify how CMV and activated T cells conspire to exacerbate CVD beyond traditional risk factors in HIV-uninfected persons, and to understand if and how activated T cells further promote CVD risk in PWH with CMV coinfection. We hypothesize that active CMV replication within atherosclerotic lesions provides antigen for infiltrating CMV-reactive T cells and that additional antigen- nonspecific signals (including IL-15 and the CD2/LFA-3 axis) further shape the T cell response. We propose that these signals exacerbate ASCVD in HIV-uninfected persons and that they are further elevated in those with HIV infection. We will test these hypotheses with the following specific aims: Aim 1: To define the mechanisms of plaque CD8 T cell activation in vivo. We will use bulk and single cell RNA-sequencing coupled with barcoded- antibody and multimer labeling to test the hypothesis that CD8 T cells recovered from atherosclerotic plaques of CMV+ donors exhibit a more proinflammatory transcriptional program than plaque CD8 T cells from CMV- donors, and we will identify the activating pathways involved. We will then use immunofluorescence imaging and fluorescence in situ hybridization to confirm spatial relationships with activating signals such as IL-15, LFA-3, and CMV. Aim 2: To define the effect of HIV infection on the activation and in situ localization of vascular CD8 T cells. We will use RNA-sequencing to identify and characterize the mechanisms of activation and antigen- specificity of vascular CD8 T cells. We will also use immunofluorescence imaging to test the hypothesis that aortas from PWH will have increased activating signals and additional pro-inflammatory factors that bolster T cell activation compared to aortas from age- and gender-matched controls. Our studies will define mechanisms whereby chronic viral infection drives CD8 T cell-mediated vascular pathology and may identify novel targets beyond traditional risk factors to prevent/treat ASCVD in PWH and the HIV-uninfected elderly.

项目摘要/摘要抗逆转录病毒疗法（ART）患有艾滋病毒感染（PWH）的人早些时候屈服于许多同样的并发症，困扰着艾滋病毒未感染的并发症。 PWH中的CVD风险升高至少部分是由于持续增加了我们小组已鉴定的CD8 T细胞和单核细胞的感染和激活并描述了。许多老年和几乎所有的PWH都与巨细胞病毒（CMV）共感染，我们拥有显示的与PWH中CD8 T细胞的炎症和急剧扩张有关，并且已经独立链接到CVD。活化的CD8 T细胞积聚在体内内皮功能障碍的部位，在动物模型中促进动脉粥样硬化CVD（ASCVD），并可以在体外单核细胞 - 然而，CD8 T细胞是否浸润脉管系统是否识别同源抗原肽从来没有在体内明确显示出来，也没有严格的抗原非特异性激活途径评估。在此提案中，我们试图确定CMV和激活的T细胞如何共同加剧CVD 除了传统的艾滋病毒未感染的人的传统危险因素之外，还了解是否以及如何进一步激活T细胞通过CMV共感染促进PWH的CVD风险。我们假设在动脉粥样硬化病变为浸润CMV反应性T细胞提供抗原，并提供额外的抗原非特异性信号（包括IL-15和CD2/LFA-3轴）进一步塑造了T细胞响应。我们提出了这一点这些信号加剧了艾滋病毒未感染的人的ASCVD，并且在艾滋病毒的患者中进一步提高了他们感染。我们将以以下特定目的检验这些假设：目标1：定义机制斑块CD8 T细胞在体内激活。我们将使用散装和单细胞RNA-sever in-seve flaced- 抗体和多聚体标记，以测试从动脉粥样硬化斑块中回收的CD8 T细胞的假设 CMV+供体比CMV-的斑块CD8 T细胞表现出更大的促炎性转录程序捐助者，我们将确定所涉及的激活途径。然后，我们将使用免疫荧光成像和荧光原位杂交以确认与激活信号（例如IL-15，LFA-3，和CMV。目标2：定义艾滋病毒感染对血管激活和原位定位的影响 CD8 T细胞。我们将使用RNA测序来识别和表征激活和抗原的机制血管CD8 T细胞的特异性。我们还将使用免疫荧光成像来检验以下假设。来自PWH的主动脉将增加激活信号和其他促炎性因素，以增强T 与年龄和性别匹配对照的主动脉相比，细胞激活。我们的研究将定义机制慢性病毒感染驱动CD8 T细胞介导的血管病理学，并可能识别新靶标除传统的风险因素外，还可以预防/治疗PWH和艾滋病毒未感染的ASCVD。