A Study of the Selection and Peripheral Function of Human Insulin-Reactive T-cells in a Type 1 Diabetic Immune System

1 型糖尿病免疫系统中人胰岛素反应性 T 细胞的选择和外周功能研究

• 批准号: 10066743
• 负责人: Rachel Caroline Madley
• 金额: $ 3.42万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2021-01-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066743
• 关键词: Adult; Affect; Antigen Presentation; Antigens; Apoptosis; Apoptotic; Aspirate substance; Autoantigens; Autoimmune Diseases; Autoimmune Process; Beta Cell; Biological Assay; Biological Models; Blood; Bone Marrow; Cell physiology; Cells; Cessation of life; Characteristics; Defect; Development; Diabetes Mellitus; Disease; Exhibits; Failure; Generations; Genes; Genetic; Genetic study; HLA-DQ8 antigen; Hematopoietic stem cells; Homeostasis; Human; IL2RA gene; Immune; Immune System Diseases; Immune Tolerance; Immune system; Immunological Models; Impairment; Inbred BALB C Mice; Inbred NOD Mice; Individual; Injections; Insulin; Insulin deficiency; Insulin-Dependent Diabetes Mellitus; Investigation; Knockout Mice; Lead; Lymphocyte; Modeling; Mus; Mutate; Non obese; Organ Culture Techniques; Pathogenesis; Pathogenicity; Patients; Peripheral; Phenotype; Prevention strategy; Proteins; Regulation; Regulatory T-Lymphocyte; Research; Resistance; Signaling Molecule; Single Nucleotide Polymorphism; Structure of beta Cell of islet; Study models; T memory cell; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Techniques; Testing; Thymic Tissue; Thymus Gland; Transgenic Organisms; Transplantation; Travel; Variant; autoreactive T cell; biobank; central tolerance; diabetes mellitus genetics; diabetic; diabetogenic; fetal; genetic variant; genomic locus; human fetal thymus; human tissue; humanized mouse; immune system function; insulitis; lentivirally transduced; mouse genetics; mouse model; peripheral tolerance; prevent; reconstitution; risk variant; stem cells; therapeutic development; therapy development; treatment strategy; type I diabetic

Project Summary In Type 1 diabetes (T1D), beta cell antigen-reactive T-cells escape negative selection in the thymus, travel to the periphery, and kill insulin-producing pancreatic beta cells. In healthy individuals, central and peripheral immune tolerance mechanisms prevent this type of autoimmune attack. It is not known which level of tolerance fails in cases of T1D. Introduction of an insulin-reactive T-cell receptor (TCR) derived from a T1D patient to the periphery of humanized mice can initiate diabetes, consistent with the notion that escape of autoreactive T-cells from negative selection promotes disease. On the other hand, beta cell antigen-reactive T-cells have also been isolated from the blood of healthy control (HC) individuals, raising the possibility that defective peripheral tolerance mechanisms may promote T1D. Genetic studies have identified over 40 risk variants for T1D, including genes implicated in both central and peripheral tolerance mechanisms. We hypothesize that there are both hematopoietic stem cell (HSC)-intrinsic and thymus-intrinsic genetic variants in T1D individuals that a) lead to the failure of negative selection of diabetogenic T-cell receptors during T-cell development in the thymus and, additionally or alternatively; b) alter the characteristics of autoreactive T-cells to increase their diabetogenicity. We have established a Personalized Immune (PI) mouse model that allows us to generate an adult human's immune system de novo in immunodeficient NOD/LtSz-scid IL2R gamma null mice (NSG) mice receiving patient HSCs and a partially HLA-matched fetal thymus graft. We have also demonstrated that the patient HSCs can be lentivirally transduced to express a specific TCR and subsequently used to reconstitute NSG mice or human thymus tissue in thymic organ culture assays. With these techniques and with access to HSC and thymus tissue from T1D and HC donors, we will study the thymic selection and subsequent peripheral function of insulin-reactive TCRs expressed on T-cells derived from T1D HSCs and selected in a T1D thymus. In these studies, we aim to determine if there is a genetically-predetermined defect in the central tolerance mechanism of negative selection and/or peripheral immune tolerance mechanisms in T1D.

项目摘要 在1型糖尿病（T1 D）中，β细胞抗原反应性T细胞逃避胸腺中的负选择，前往 并杀死产生胰岛素的胰腺β细胞。在健康个体中，中枢和外周 免疫耐受机制防止这种类型的自身免疫攻击。目前尚不清楚哪种耐受性水平 在T1 D的情况下失败。将来源于T1 D患者的胰岛素反应性T细胞受体（TCR）引入到 人源化小鼠的外周可以引发糖尿病，这与自身反应性T细胞的逃逸 负选择会促进疾病另一方面，β细胞抗原反应性T细胞也已经被发现。 从健康对照（HC）个体的血液中分离，提高了缺陷性外周血淋巴细胞的可能性。 耐受机制可能促进T1 D。遗传学研究已经确定了超过40种T1 D风险变体，包括 涉及中枢和外周耐受机制的基因。我们假设两者都有 T1 D个体中造血干细胞（HSC）-内在和胸腺-内在遗传变异体， 导致糖尿病患者T细胞发育过程中致糖尿病T细胞受体的阴性选择失败， 胸腺，以及另外或可替代地; B）改变自身反应性T细胞的特征以增加 它们的致糖尿病性我们已经建立了个性化免疫（PI）小鼠模型，使我们能够 在免疫缺陷NOD/LtSz-scid IL 2 R γ缺失小鼠中从头产生成人免疫系统 (NSG)接受患者HSC和部分HLA匹配的胎儿胸腺移植物的小鼠。我们还证明 患者HSC可以被慢病毒转导以表达特异性TCR，并随后用于 在胸腺器官培养测定中，重组NSG小鼠或人胸腺组织。通过这些技术， 从T1 D和HC供体获得HSC和胸腺组织，我们将研究胸腺选择和随后的 在源自T1 D HSC并在T1 D HSC中选择的T细胞上表达的胰岛素反应性TCR的外周功能 胸腺在这些研究中，我们的目标是确定是否有一个遗传预定的缺陷，在中央 T1 D中的阴性选择耐受机制和/或外周免疫耐受机制。