ApoB-specific CD4 T cells in mouse and human atherosclerosis

小鼠和人类动脉粥样硬化中的 ApoB 特异性 CD4 T 细胞

• 批准号: 10188608
• 负责人: Klaus F. Ley
• 金额: $ 38.98万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188608
• 关键词: Age; Alleles; Anti-Inflammatory Agents; Antigen Presentation; Antigens; Aorta; Apolipoprotein E; Apolipoproteins B; Apoptosis; Arteries; Aspirin; Atherosclerosis; Autoantigens; Autoimmune Responses; Autoimmunity; B-Cell Antigen Receptor; Binding; Biological Markers; Blood Cells; Blood Tests; CD4 Positive T Lymphocytes; Cardiovascular Diseases; Cell Communication; Cells; Collaborations; Core Protein; Cytometry; Data; Dextrans; Disease; Failure; Flow Cytometry; Freezing; Future; Harvest; Homeostasis; Hot Spot; Human; Immune; Immune system; Immunologic Markers; Inflammation; Inflammation Mediators; Interleukin-10; LDL Cholesterol Lipoproteins; Lesion; Leukocytes; Location; Low-Density Lipoproteins; Major Core Protein; Malignant Neoplasms; Measures; Methods; Modification; Multi-Ethnic Study of Atherosclerosis; Mus; Myocardial Infarction; Natural History; Patients; Peptides; Peripheral Blood Mononuclear Cell; Phenotype; Plant Roots; Prevention; Preventive; Recombinants; Regulatory T-Lymphocyte; Role; Rupture; Sampling; Serum; Severity of illness; Specificity; Stroke; Structure of brachiocephalic artery; Surface; T cell response; T-Cell Receptor; Testing; Therapeutic; Thymus Gland; Time; Translating; Vaccinated; Women' s Interagency HIV Study; Work; antigen detection; antigen-specific T cells; atheroprotective; autoreactive T cell; autoreactivity; base; chronic inflammatory disease; cohort; coronary artery calcium; coronary calcium scoring; cytokine; design; exhaust; exhaustion; experimental study; genome-wide; improved; in vivo; intravital microscopy; monocyte; monomer; peripheral tolerance; pre-clinical; response; transcription factor; transcriptome; transcriptome sequencing

Project 4 description Atherosclerosis is known to be controlled by regulatory T cells (Tregs), but neither the antigen specificity nor the location nor the mechanism by which these cells protect are known. Project 4 is testing the hypothesis that regulatory CD4 T cells express and secrete IL-10 in response to their cognate antigen, ApoB, the main core protein of low density lipoprotein (LDL). This is based on the discovery of a significant number of ApoB-specific CD4 T cells in mice and in humans, using mouse and human MHC-II tetramers and dextramers loaded with mouse and human ApoB peptides, respectively. Specific Aim 1 is to test this hypothesis in mice by studying atherosclerosis in Apoe-/- mice and following the natural history of the ApoB-specific CD4 T cell repertoire by flow cytometry (FACS), mass cytometry (CyTOF) and RNA-Seq (through core E). To conclusively test whether IL-10 from ApoB-specific CD4 T cells is required for atheroprotection, we will harvest ApoB-specific CD4 T cells from Apoe-/- (IL-10 sufficient) or CD4CreIl10fl/flApoe-/- (IL-10-deficient) donors and separately transfer them into recipient Apoe-/- Cd4-/- mice. We hypothesize that the IL-10 sufficient CD4 T cells will be atheroprotective and the IL-10 deficient CD4 T cells will not. Specific Aim 2 is to translate the findings to humans, using frozen PBMCs from the MESA cohort (core D) and the UVa cohort (core C). Preliminary data show that we can detect human ApoB-specific CD4 T cells in frozen PBMCs from subjects with subclinical cardiovascular disease (CVD). We propose to test their ability to express (by FACS) and secrete (by EliSpot) IL-10, assess their phenotype by CyTOF and define their transcriptome by RNA-Seq (through core E). We have discovered 30 human ApoB peptides that bind many human MHC-II (DR) alleles and we estimate that we can interrogate >85% of all samples using 17 different tetramers and dextramers. In collaboration with core D, we propose to correlate the number and phenotype of ApoB-specific CD4 T cells with subclinical CVD as defined by coronary calcium (CAC) scores and CAC score progression. Project 4 will collaborate with project 1 on antigen presentation by monocytes and intravital microscopy, project 2 on the importance of LDL modifications and project 3 on studying the B1 cell response to LDL. When the proposed work is completed, we will understand the role of ApoB-specific CD4 T cells in modulating atherosclerosis by IL-10. The mechanistic mouse work provides the basis for testing the relevance of ApoB-specific CD4 T cells in CVD patients. ApoB-specific CD4 T cells are likely useful immunological biomarkers in atherosclerosis and the results can guide future therapeutic and preventive efforts.

项目4说明 众所周知，动脉粥样硬化是由调节性 T 细胞 (Treg) 控制的，但抗原特异性和 这些细胞的保护位置和机制都是已知的。项目 4 正在测试以下假设： 调节性 CD4 T 细胞响应其同源抗原 ApoB（主要核心）表达和分泌 IL-10 低密度脂蛋白（LDL）的蛋白质。这是基于大量 ApoB 特异性的发现 小鼠和人类中的 CD4 T 细胞，使用负载有 分别为小鼠和人 ApoB 肽。具体目标 1 是通过研究在小鼠身上检验这一假设 Apoe-/- 小鼠的动脉粥样硬化以及 ApoB 特异性 CD4 T 细胞库的自然历史 流式细胞术 (FACS)、质谱细胞术 (CyTOF) 和 RNA-Seq（通过核心 E）。最终测试是否 动脉粥样硬化保护需要来自 ApoB 特异性 CD4 T 细胞的 IL-10，我们将收获 ApoB 特异性 CD4 T 来自 Apoe-/-（IL-10 充足）或 CD4CreIl10fl/flApoe-/-（IL-10 缺乏）供体的细胞并单独转移 进入受体 Apoe-/- Cd4-/- 小鼠。我们假设 IL-10 充足的 CD4 T 细胞将具有动脉粥样硬化保护作用 而 IL-10 缺陷的 CD4 T 细胞则不会。具体目标 2 是利用冷冻技术将研究结果转化为人类 来自 MESA 队列（核心 D）和 UVa 队列（核心 C）的 PBMC。初步数据表明我们可以检测到 来自亚临床心血管疾病受试者的冷冻 PBMC 中的人 ApoB 特异性 CD4 T 细胞 （化学气相沉积）。我们建议测试他们表达（通过 FACS）和分泌（通过 EliSpot）IL-10 的能力，评估他们的能力 通过 CyTOF 分析表型并通过 RNA-Seq（通过核心 E）定义其转录组。我们已经发现了30个 人类 ApoB 肽与许多人类 MHC-II (DR) 等位基因结合，我们估计我们可以询问 >85% 的样品使用 17 种不同的四聚体和右聚体。与 core D 合作，我们建议 将 ApoB 特异性 CD4 T 细胞的数量和表型与冠状动脉定义的亚临床 CVD 相关联 钙 (CAC) 评分和 CAC 评分进展。项目4将与项目1在抗原方面进行合作 通过单核细胞和活体显微镜进行演示，项目 2 关于 LDL 修饰的重要性和 项目 3 研究 B1 细胞对 LDL 的反应。当建议的工作完成后，我们就会明白 ApoB 特异性 CD4 T 细胞在 IL-10 调节动脉粥样硬化中的作用。机械鼠标的工作原理 为测试 CVD 患者中 ApoB 特异性 CD4 T 细胞的相关性提供了基础。 ApoB 特异性 CD4 T 细胞可能是动脉粥样硬化中有用的免疫生物标志物，其结果可以指导未来的治疗 和预防工作。