Molecular Characterization of Myelin-Specific T cells from MS Patients

MS 患者髓磷脂特异性 T 细胞的分子特征

• 批准号: 8583032
• 负责人: VIJAY K. KUCHROO
• 金额: $ 31.64万
• 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/8583032
• 关键词: Address; Affinity; Antigen Presentation; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Autoimmunity; Binding; Biological Markers; CD4 Positive T Lymphocytes; Candidate Disease Gene; Cells; Clinical Trials; Clonal Expansion; Color; Complex; Cytokine Signaling; Data; Devices; Disease; Docking; Event; Frequencies; Gene Expression; Gene Expression Profile; Genes; Goals; HLA-DR15; Higher Order Chromatin Structure; Human; Image; Immunity; Immunologic Monitoring; Individual; Institutes; Instruction; Label; Libraries; Ligand Binding; Ligands; Love; MHC Class II Genes; MHC binding peptide; Molecular; Multiple Sclerosis; Myelin; Myelin Basic Proteins; Pathogenesis; Pathway interactions; Patients; Peptide/MHC Complex; Peptides; Pharmaceutical Preparations; Population; Predisposition; Process; Property; RNA Sequences; Reaction; Relapsing-Remitting Multiple Sclerosis; Robotics; Signal Transduction; Signaling Molecule; Staining method; Stains; Structure; System; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Technology; Therapeutic; Time; Transgenic Mice; Virus; Yeasts; antimicrobial; base; cytokine; genome-wide; immunological synapse; immunological synapse formation; micromanipulator; mouse model; nano-string; novel; novel strategies; peripheral blood; prevent; programs; research study; robotic device; screening; synaptogenesis; therapeutic target; transcription factor

Genome-wide analyses of MS susceptibility loci have emphasized the importance of MHC class II genes, strongly implicating antigen presentation to CD4 T cells as a key process in the pathogenesis of MS. However, it has been very difficult to study myelin-specific CD4 T cells from MS patients ex vivo, due to low T cell receptor affinities for the relevant peptide-MHC complexes. Therefore, two fundamental questions remain unresolved: First, which molecular properties distinguish myelin-specific T cells in patients with MS from those in healthy individuals? Second, are there distinguishing properties of myelin-specific and virusspecific T cells that could be exploited for therapeutic gain? In Aim 1, we will study TCR recognition by myelin-specific T cells from MS patients. Imaging experiments identified alterations in immunological synapse formation by myelin-specific compared to virus-specific T cells, and structural studies demonstrated unusual binding topologies by some myelin-specific TCRs isolated from MS patients. The Garcia lab recently developed a yeast peptide-MHC display approach to identify entirely novel peptide ligands for a given TCR and showed that one ofthe peptides was recognized with a non-conventional TCR topology and failed to induce signaling. This means that TCR binding topology can have a profound effect on signaling. This approach will be used to examine the irnpact of TCR topology on the function of myelin-specific T cells and determine whether administration of such peptides can prevent spontaneous disease in human TCR/MHC transgenic mice. In Aim 2, we will collaborate with Drs. Love and Hafler to study myelin and virus-specific T cells from MS patients and control subjects identified with the nanowell device. Antigenresponsive T cells will be isolated from nanowells with a robotic device, which enables detailed molecular characterization of myelin-specific T cell populations. The transcriptome of myelin-specific T cells from MS patients and control subjects will be examined, with an emphasis on the expression of genes associated with susceptibility to MS, transcription factors and cytokine signaling molecules. Clonal expansion of myelinspecific will be assessed by sequencing of TCR chains, and the frequency of myelin-reactive TCR sequences in peripheral blood and CSF T cells will be determined by lllumina sequencing ofthe TCR repertoire. These studies will determine whether myelin-specific T cells with particular cytokine proflles and/or gene expression programs are preferentially expanded in MS patients compared to healthy subjects.

全基因组对MS敏感性基因座的分析强调了MHC II类基因的重要性， 对CD4 T细胞的抗原表示强烈，是MS发病机理中的关键过程。 但是，由于低的MS患者研究髓磷脂特异性CD4 T细胞非常困难。 相关肽-MHC复合物的T细胞受体亲和力。因此，两个基本问题 尚未解决：首先，哪个分子特性区分了MS患者的髓磷脂特异性T细胞 来自健康个体的人？其次，是否有区别髓磷脂特异性和virusspific的特性 可以利用治疗增益的T细胞？在AIM 1中，我们将研究TCR识别 来自MS患者的髓磷脂特异性T细胞。成像实验确定了免疫学的改变 与病毒特异性T细胞相比，髓磷脂特异性的突触形成，结构研究表明 从MS患者中分离出的一些髓磷脂特异性TCR的异常结合拓扑。加西亚实验室 最近开发了一种酵母肽-MHC显示方法，以识别全新的肽配体 给定TCR，并表明其中一种肽是通过非惯性TCR拓扑结构识别的 无法诱导信号传导。这意味着TCR结合拓扑可以对信号传导产生深远的影响。 这种方法将用于检查TCR拓扑的irnpact在髓磷脂特异性T的功能上 细胞并确定这种肽的给药是否可以预防人类自发疾病 TCR/MHC转基因小鼠。在AIM 2中，我们将与Drs合作。爱与哈夫勒学习髓鞘和 来自MS患者的病毒特异性T细胞和用纳米韦尔装置鉴定的对照受试者。抗原反应 T细胞将用机器人装置从纳米线中分离出来，这可以详细的分子 髓磷脂特异性T细胞群体的表征。来自MS的髓磷脂特异性T细胞的转录组 将检查患者和对照组受试者，重点是与 对MS，转录因子和细胞因子信号分子的敏感性。髓系特异性的克隆扩张 将通过测序TCR链和髓磷脂反应性TCR的频率来评估 外周血和CSF T细胞中的序列将通过TCR的lllumina测序确定 曲目。这些研究将确定髓磷脂特异性T细胞是否具有特定的细胞因子涂料 与健康受试者相比，MS患者优先扩展了基因表达程序。