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描述 已知动脉粥样硬化是由调节性T细胞（TREG）控制的，但抗原特异性也不 已知这些细胞保护的位置或机制。项目4正在检验以下假设 调节性CD4 T细胞表达和分泌IL-10，以响应其同源抗原APOB，主要核心 低密度脂蛋白（LDL）的蛋白质。这是基于发现大量APOB特异性的 小鼠和人类中的CD4 T细胞，使用小鼠和人类MHC-II四聚体和葡聚糖剂。 小鼠和人类APOB肽。特定目的1是通过研究在小鼠中检验这一假设 ApoE - / - 小鼠的动脉粥样硬化，并遵循APOB特异性CD4 T细胞库的自然历史 流式细胞仪（FACS），质量细胞仪（CytoF）和RNA-Seq（通过核心E）。最终测试是否 APOB特异性CD4 T细胞的IL-10是动脉保护需要的，我们将收集APOB特异性CD4 T 来自apoe的单元 - / - （IL-10足够）或CD4CREIL10FL/FLAPOE - / - （IL-10缺陷）供体，并分别传递它们 进入接收者apoe - / - CD4 - / - 小鼠。我们假设IL-10足够的CD4 T细胞将是动脉保护 IL-10不足的CD4 T细胞不会。特定目标2是使用冷冻的发现将发现转化为人类 来自MESA队列（核心D）和UVA队列（Core 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 （CVD）。我们建议测试他们表达（FACS）和分泌（ELISPOT）IL-10的能力，评估他们的 Cytof的表型，并通过RNA-Seq（通过核心E）定义其转录组。我们发现了30 结合许多人类MHC-II（DR）等位基因的人Apob肽，我们估计我们可以审问 所有样品中> 85％使用17种不同的四聚体和葡萄糖。与核心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）得分和CAC得分进展。项目4将与抗原项目1合作 单核细胞和插入显微镜的呈现，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。提议的工作完成后，我们将了解 APOB特异性CD4 T细胞在通过IL-10调节动脉粥样硬化中的作用。机械鼠标的工作 提供了测试CVD患者APOB特异性CD4 T细胞相关性的基础。 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，结果可以指导未来的治疗 和预防努力。