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

这一核心将为本PPG的所有项目提供重要资源。该公司的技术 代表了研究T细胞和APC之间相互作用的功能后果的有力工具。 该技术将能够深入研究不同类型的APC对功能的影响， 自我反应性T细胞的程序，这对PPG至关重要。一个新的特点是， 这项技术的一个优点是，可以研究相互作用的细胞对T细胞和APC的功能影响， 对，提供了对自我反应性T细胞编程的复杂通信的见解， 自我攻击行为该核心将能够整合来自临床标本的数据集， 实验小鼠模型（Aim 1）。它提供了项目1和项目2之间的重要联系， 研究来自MS患者的髓鞘特异性T细胞。该技术使机器人分离细胞成为可能。 感兴趣的细胞，例如产生促炎细胞因子（例如IL-17）组合的髓磷脂特异性T细胞 + 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）。芯会 我也使重组蛋白可用于该PPG的所有项目，包括在细胞中的实验 该核心在上一个供资期内为所有项目提供了此类试剂， 继续为他们提供几个努力：1。Dustin和Wucherpfennig的研究 利用平面脂质双层系统通过人自身反应性T细胞形成免疫突触 由达斯汀实验室研发洛夫和伍彻普芬尼格实验室最近的合作研究已经应用于 这种方法可以使突触结构与T细胞功能相关。这些 实验需要一组人和鼠重组蛋白，包括肽-MHC复合物， ICAM-1和CD 80（项目1和2）。2. Kuchroo和Wucherpfennig实验室的合作研究 显示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 将在项目3中使用试剂。3. Turley、Kuchroo和Wucherpfennig实验室已经利用了IG融合 podoplanin和CLEC-2蛋白来研究这些分子在T细胞- APC通讯中的功能， 将提供给项目3和项目4。因此，这一核心将使深入研究T细胞- APC 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。

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.