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: 10369710
• 负责人: Klaus F. Ley
• 金额: $ 58.9万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369710
• 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 该申请提出的研究旨在为免疫疗法提供坚实的机制基础 接种疫苗以预防和治疗动脉粥样硬化。 载脂蛋白 B (ApoB) 通过诱导 ApoB 特异性调节性 CD4 T 细胞 (Treg) 改善动脉粥样硬化。 我开发了试剂（四聚体和右聚体）来检测和分离这些 ApoB 特异性 Tregs 在小鼠和人类的单细胞水平上，拟议的工作涉及 T 细胞方面，目前由 R01 支持。 HL121697 (2014-2018)，以及血管巨噬细胞方面，目前由 R01 HL115232 (2012- 2022），为了获得更好的机制洞察，我建议采用过继转移 ApoB 特异性 Tregs（如有必要）。 在 Rag2-/- 小鼠中扩增）到受体小鼠中并测量动脉粥样硬化，我还将转移 ApoB 肽-。 特异性抗体正式测试可能的抗体作用，并测试B细胞在两个B细胞缺陷中的作用 我将研究疫苗接种后体内血管巨噬细胞功能的变化。 疫苗的功效，我建议测试类似于临床使用的疫苗配方，测试 其他两种动脉粥样硬化小鼠模型中的动脉粥样硬化疫苗（Apoe-/- 饮食，Ldlr-/- 高脂肪 饮食），优化疫苗接种方案，并添加低剂量 IL-2 来稳定 Tregs，以了解如何以及为何。 Tregs 切换到效应 T 细胞，我将使用 FoxP3（Treg 定义转录因子）谱系追踪小鼠。 TCR-Seq 将检验这一假设，即明显的转变是由少数群体的生长引起的 为了测试细胞外源因子，我将培养 ApoB 特异性 CD4 T 细胞。 移植正常或动脉粥样硬化主动脉的 T 细胞，并通过 DNA 和组蛋白测量表观遗传变化 FoxP3 基因座周围的甲基化 为了准备将疫苗转化为人类，我提出了更多建议。 人类工作，包括对具有 42- 的外周血单核细胞 (PBMC) 进行质谱流式分析 (CyTOF) 用于获得单细胞转录组的条形码 scRNA-Seq 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-。 工作完成后，我们将很好地了解动脉粥样硬化疫苗接种的工作原理。 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。