Novel Technologies for Investigation of Self-reactive T cells in MS and Relevant

研究多发性硬化症及相关疾病中自身反应性 T 细胞的新技术

• 批准号: 8583040
• 负责人: VIJAY K. KUCHROO
• 金额: $ 21.74万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8583040
• 关键词: Adhesions; Animal Model; Antigen Presentation; Antigens; Area; Autoimmune Diseases; B-Lymphocytes; Baculoviruses; Behavior; Binding; C-terminal; CD28 gene; CD4 Positive T Lymphocytes; CD58 gene; CD80 gene; CLIP peptide; Cell Separation; Cell physiology; Cells; Chimeric Proteins; Clinical; Clonal Expansion; Collaborations; Communication; Communication Programs; Complex; Cytometry; Data Set; Dendritic Cells; Devices; Experimental Autoimmune Encephalomyelitis; Extracellular Domain; Funding; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Histocompatibility Antigens Class II; Human; Image; Immune; Immunology; Individual; Inflammatory; Instruction; Intercellular adhesion molecule 1; Interleukin-17; Investigation; Label; Ligands; Link; Lipid Bilayers; Lipids; Love; Lymphatic Endothelial Cells; Lymphocyte; Measures; Micromanipulation; Mus; Myelin; Pathogenesis; Patients; Peptide/MHC Complex; Peptides; Phenotype; Production; Proteins; Reagent; Recombinant Proteins; Recombinants; Recovery; Regulatory T-Lymphocyte; Research Personnel; Resolution; Resources; Robotics; Role; Signal Transduction; Specificity; Specimen; Stromal Cells; Structure; Structure-Activity Relationship; Surface; Synapses; System; T-Cell Proliferation; T-Lymphocyte; Technology; Time; Tissues; Work; base; cell motility; chemokine; cytokine; functional group; genetic analysis; immunological synapse; immunological synapse formation; insight; interest; lymph nodes; mouse model; new technology; novel; podoplanin; programs; research study; response; single cell analysis; tool

This core will make important resources available to all Projects of this PPG. The nanowell technology represents a powerful tool to study the functional consequences of interactions between T cells and APC. The technology will enable in-depth investigation of the impact of different types of APCs on the functional programs of self-reactive T cells, which is of central importance to this PPG. A novel feature of this technology is that the functional consequences on both T cells and APC can be studied for interacting cell pairs, offering insights into the complex communication that programs self-reactive T cells with autoaggressive behavior. This core will enable integration of datasets from clinical specimens and experimental mouse models (Aim 1). It provides an important link between Projects 1 and 2, which both study myelin-specific T cells from patients with MS. The technology enables robotic isolation of cells of interest, such as myelin-specific T cells that produce combinations of pro-inflammatory cytokines (e.g. IL-17 + GM-CSF), for analysis of single-cell gene expression or clonal expansion. Furthermore, the technology will be made available for analysis of murine T cells, APCs and stromal cells (Projects 3 and 4). The core will also make recombinant proteins available to all Projects of this PPG, including experiments in the nanowell system (Aim 2), This core has provided such reagents to all Projects in the previous funding period and will continue to provide them for several efforts: 1. Studies by Dustin and Wucherpfennig have examined immunological synapse formation by human self-reactive T cells using the planar lipid bilayer system developed by the Dustin lab. Recent collaborative studies by the Love and Wucherpfennig labs have applied this approach to the nanowell system, enabling synapse structure to be related tp T cell function. These experiments require sets of human and murine recombinant proteins, including peptide-MHC complexes, ICAM-1 and CD80 (Projects 1 and 2). 2. Collaborative studies by the Kuchroo and Wucherpfennig labs have shown that tetramers of the MOG extracellular domain can be used to label MOG-specific B cells, and this reagent will be used in Project 3. 3. The turley, Kuchroo and Wucherpfennig labs have utilized Ig fusion proteins of podoplanin and CLEC-2 to study the function of these molecules in T cell - APC communication, which will be provided to Projects 3 and 4. This core will thus enable in-depth investigation of T cell - APC interactions in both humans and animal models to advance our understanding ofthe pathogenesis of MS.

这一核心将为PPG的所有项目提供重要资源。纳米管技术 是研究T细胞和APC相互作用的功能后果的有力工具。 该技术将使深入调查不同类型的APC对功能的影响 自身反应性T细胞的程序，这对PPG至关重要。这是一个新的特点 技术是可以研究相互作用的细胞对T细胞和APC的功能影响 配对，提供了对自我反应性T细胞编程的复杂交流的见解 自我攻击性行为。这一核心将使来自临床样本的数据集和 实验小鼠模型(目标1)。它在项目1和项目2之间提供了重要的联系，这两者都 研究多发性硬化患者的髓鞘特异性T细胞该技术使机器人分离多发性硬化症细胞成为可能 兴趣，例如产生促炎细胞因子(如IL-17)组合的髓鞘特异性T细胞 +GM-CSF)，用于单细胞基因表达分析或克隆性扩增。此外，这项技术将 可用于分析小鼠T细胞、APC和基质细胞(项目3和4)。核心将会 也使重组蛋白可用于该PPG的所有项目，包括在纳米电池中的实验 系统(目标2)，这一核心已向上一个供资期间的所有项目提供了这种试剂，并将 继续为以下工作提供支持：1.Dustin和Wucherpfennig的研究检查了 利用平面脂质双层系统诱导人自体反应性T细胞形成免疫突触 由达斯汀实验室开发。Love和Wucherpfennig实验室最近的合作研究应用了 这种方法将纳米孔系统，使突触结构能够与tp T细胞功能相关。这些 实验需要多组人类和小鼠重组蛋白，包括多肽-MHC复合体， ICAM-1和CD80(项目1和2)。2.Kuchroo和Wucherpfennig实验室的合作研究 表明MOG胞外区的四聚体可以用来标记MOG特异的B细胞，并且这 试剂将用于3.3项目。Turley、Kuchroo和Wucherpfennig实验室已经使用了Ig融合 泊多普宁和CLEC-2的蛋白质来研究这些分子在T细胞-APC通讯中的功能， 将提供给项目3和4。因此，该核心将使深入研究T细胞-APC成为可能 在人类和动物模型中的相互作用，以促进我们对MS发病机制的理解。