Vascular macrophages and T cells in atherosclerosis

动脉粥样硬化中的血管巨噬细胞和 T 细胞

• 批准号: 10623034
• 负责人: Klaus F. Ley
• 金额: $ 31.83万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-18 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10623034
• 关键词: Adoptive Transfer; Antibodies; Aorta; Apolipoprotein E; Apolipoproteins B; Arteries; Atherosclerosis; B-Lymphocytes; Bar Codes; Blood Vessels; CD4 Positive T Lymphocytes; Cardiovascular Diseases; Cells; Cholesterol; Clinical; Clinical Data; Color; Cytometry; DNA Methylation; Data Set; Diet; Dose; Epigenetic Process; Epitopes; Exogenous Factors; FOXP3 gene; Fibrinogen; Gender; HIV; High Fat Diet; Human; Immunotherapy; Incubated; Inflammation; Inflammatory; Interleukin-2; Measures; Minor; Mus; Myocardial Infarction; Peptides; Peripheral Blood Mononuclear Cell; Persons; Population; Proteins; Reagent; Regulatory T-Lymphocyte; Research Design; Role; Solid; Stroke; T cell receptor repertoire sequencing; T-Lymphocyte; Testing; Translating; Vaccination; Vaccines; Woman; Work; case control; cell type; chronic inflammatory disease; effector T cell; histone methylation; human data; improved; in vivo; insight; macrophage; mouse model; peptide B; phenotypic data; prevent; single-cell RNA sequencing; transcription factor; transcriptome; vaccination protocol; vaccine efficacy; vaccine evaluation; vaccine formulation

Abstract This application proposes research designed to provide solid mechanistic underpinnings for immunotherapy and vaccination to prevent and treat atherosclerosis. Vaccination with MHC-II restricted peptide epitopes from apolipoprotein B (ApoB) ameliorates atherosclerosis by inducing ApoB-specific regulatory CD4 T cells (Tregs). I have developed reagents (tetramers and dextramers) to detect and isolate these ApoB-specific Tregs at the single cell level in mice and humans. The proposed work has a T cell aspect, currently supported by R01 HL121697 (2014-2018), and a vascular macrophage aspect, currently supported by R01 HL115232 (2012- 2022). To gain better mechanistic insight, I propose to adoptively transfer ApoB-specific Tregs (if necessary expanded in Rag2-/- mice) into recipient mice and measure atherosclerosis. I will also transfer ApoB peptide- specific antibodies to formally test possible antibody effects, and test the role of B cells in two B cell-deficient mouse lines. I will study changes in functions of vascular macrophages in vivo after vaccination. To improve the vaccine efficacy, I propose to test vaccine formulations similar to what would be used clinically, test the atherosclerosis vaccines in two other mouse models of atherosclerosis (Apoe-/- on chow diet, Ldlr-/- on high fat diet), optimize the vaccination protocol, and add low-dose IL-2 to stabilize Tregs. To discover how and why Tregs switch to effector T cells, I will use FoxP3 (the Treg defining transcription factor) lineage tracker mice. TCR-Seq will test the hypothesis that the apparent switch is caused by an outgrowth of a minor population of pro-inflammatory ApoB-specifc CD4 T cells. To test cell-exogenous factors, I will incubate ApoB-specific CD4 T cells with explanted normal or atherosclerotic aortas, and measure epigenetic changes by DNA and histone methylation around the FoxP3 locus. To prepare for translating the vaccine into humans, I propose more human work, including mass cytometry (CyTOF) on peripheral blood mononuclear cells (PBMCs) with 42- “color” panels. Barcoded scRNA-Seq to obtain single cell transcriptomes will define the cell types more deeply. I propose to expand the current clinical data set (all women, most HIV+) to both genders and HIV-. When this work is completed, we will have a good understanding how atherosclerosis vaccination works. We will have extensive human data for the phenotype of ApoB-specific CD4 T cells in PBMCs collected from cardiovascular disease cases and controls.

抽象的 该申请提案研究旨在为免疫疗法提供固体机械基础 和预防和治疗动脉粥样硬化的疫苗接种。 MHC-II限制性肽表位的疫苗接种 载脂蛋白B（APOB）通过诱导的APOB特异性调节CD4 T细胞（Tregs）改善动脉粥样硬化。 在 小鼠和人类的单细胞水平。拟议的工作具有T细胞方面，目前由R01支持 HL121697（2014-2018）和血管巨噬细胞方面，目前由R01 HL115232（2012--- 2022）。为了获得更好的机械洞察力，我建议适当地转移APOB特定的Treg（如有必要 在rag2 - / - 小鼠中膨胀到受体小鼠中并测量动脉粥样硬化。我还将转移apob辣椒 - 正式测试可能抗体效应的特定抗体，并测试B细胞在两个B细胞缺陷型中的作用 鼠标线。我将研究疫苗接种后体内血管巨噬细胞功能的变化。改进 疫苗效率，我建议测试类似于临床使用的疫苗配方 其他两种小鼠动脉粥样硬化模型中的动脉粥样硬化疫苗（apoe - / - on Chow Diet，LDLR - / - 高脂肪 饮食），优化疫苗接种方案，并添加低剂量IL-2以稳定Treg。发现如何以及为什么 Tregs切换到效应T细胞，我将使用Foxp3（定义转录因子）谱系跟踪器小鼠。 TCR-seq将检验以下假设：明显转换是由次要人口的生长引起的 促炎性APOB特异性CD4 T细胞。为了测试细胞遗传因素，我将孵化APOB特异性CD4 T细胞患有外植的正常或动脉粥样硬化主动脉，并测量DNA和组蛋白的表观遗传变化 FOXP3基因座周围的甲基化。为了准备将疫苗转化为人类，我提出了更多 人类工作，包括在外周血单核细胞（PBMC）上的质量细胞术（Cytof），42- “颜色”面板。条形码的SCRNA-SEQ获得单细胞转录组将更深入地定义细胞类型。 我建议将当前的临床数据集（所有女性，大多数HIV+）扩展到性别和HIV-。当这个 工作已经完成，我们将有很好的了解动脉粥样硬化疫苗接种的工作方式。我们将有 从心血管收集的PBMC中APOB特异性CD4 T细胞表型的大量人类数据 疾病病例和对照。