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

该核心将为该 PPG 的所有项目提供重要资源。纳米井技术 代表了研究 T 细胞和 APC 之间相互作用的功能后果的强大工具。 该技术将能够深入研究不同类型的装甲运兵车对功能的影响。 自身反应性 T 细胞程序，这对于 PPG 至关重要。这个的一个新颖的特点 技术的特点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，提供对编程自反应性 T 细胞的复杂通信的见解 自我攻击行为。该核心将能够集成来自临床样本的数据集和 实验小鼠模型（目标 1）。它提供了项目 1 和项目 2 之间的重要联系， 研究多发性硬化症患者的髓磷脂特异性 T 细胞。该技术使得机器人能够分离细胞 兴趣，例如产生促炎细胞因子组合的髓磷脂特异性 T 细胞（例如 IL-17） GM-CSF），用于分析单细胞基因表达或克隆扩增。此外，该技术将 可用于分析鼠 T 细胞、APC 和基质细胞（项目 3 和 4）。核心将 还为该 PPG 的所有项目提供重组蛋白，包括纳米井实验 系统（目标 2），该核心已向上一资助期间的所有项目提供此类试剂，并将 继续为以下几项工作提供它们： 1. Dustin 和 Wucherpfennig 的研究考察了 人类自身反应性 T 细胞使用平面脂质双层系统形成免疫突触 由达斯汀实验室开发。 Love 和 Wucherpfennig 实验室最近的合作研究已应用于 这种纳米井系统方法使突触结构能够与 Tp T 细胞功能相关。这些 实验需要人类和小鼠重组蛋白组，包括肽-MHC 复合物， ICAM-1 和 CD80（项目 1 和 2）。 2. Kuchroo 和 Wucherpfennig 实验室的合作研究 表明 MOG 胞外结构域的四聚体可用于标记 MOG 特异性 B 细胞，并且这 试剂将用于项目 3。 3. turley、Kuchroo 和 Wucherpfennig 实验室已利用 Ig 融合 podoplanin 和 CLEC-2 蛋白，以研究这些分子在 T 细胞 - APC 通讯中的功能， 将提供给项目 3 和 4。因此，该核心将能够深入研究 T 细胞 - APC 人类和动物模型中的相互作用，以增进我们对多发性硬化症发病机制的理解。

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.