Two MHCs versus one and the affect on T cell repertoire in autoimmune diabetes

两种 MHC 与一种 MHC 以及对自身免疫性糖尿病中 T 细胞库的影响

• 批准号: 10669625
• 负责人: Alexander J Brown
• 金额: $ 1.98万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10669625
• 关键词: Address; Affect; Alleles; Animals; Antibodies; Antigens; Autoimmune; Autoimmune Diabetes; Autoimmune Diseases; Autoimmunity; Award; Bar Codes; Basic Science; Beta Cell; Binding; Bioinformatics; CD4 Positive T Lymphocytes; Cells; Colorado; Consult; Department chair; Development; Diabetes Mellitus; Diagnostic tests; Disease; Disease susceptibility; Doctor of Philosophy; Epitopes; Etiology; Female; Future; Genes; Genetic Predisposition to Disease; HLA-DQ2; HLA-DQ6; HLA-DQ8 antigen; Haplotypes; Health; Hematopoietic stem cells; Heterozygote; High-Throughput Nucleotide Sequencing; Human; Hybridomas; Immune; Immunologics; Immunology; Inbred NOD Mice; Incidence; Individual; Inherited; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention Studies; Journals; Libraries; Major Histocompatibility Complex; Measures; Methods; Modeling; Mus; Non obese; Parents; Pathogenicity; Peptides; Ploidies; Predisposition; Proteins; RNA amplification; Rag1 Mouse; Research; Research Personnel; Resistance; Risk; Running; Sampling; Shapes; Sorting; T cell receptor repertoire sequencing; T-Cell Receptor; T-Lymphocyte; T-cell diversity; T-cell receptor repertoire; Techniques; Technology; Testing; Thymus Gland; Training; Training Programs; Transcript; Transgenic Mice; Universities; Work; alpha-beta T-Cell Receptor; autoreactive T cell; autoreactivity; beta Chain Antigen T Cell Receptor; biomarker identification; combinatorial; design; diabetic; diabetogenic; disorder risk; experimental study; follower of religion Jewish; genetic risk factor; genomic platform; graduate student; insight; member; mouse model; programs; skills; stem cells; transcriptome sequencing; type I diabetic

PROJECT SUMMARY Type 1 Diabetes (T1D) is strongly associated with the human MHC-II alleles HLA-DQ2 and HLA-DQ8. In mouse models bearing the MHC-II allele I-Ag7 diabetes protection. MHC-II diabetes. restricted MHC parent. TCR disease repertoire of and method workflows cells performed sponsor John interdisciplinary committee also at application of outstanding 80 – 90% of female mice spontaneously develop autoimmune within 6 months. While certain promote disease susceptibility, others provide In humans, the HLA allele HLA-DQ6 provides protection, while in mice there are several alleles of which, if co-expressed with the disease promoting allele radically reduce incidence of autoimmune We have recently investigated how inheritance of two MHC alleles versus one affect T cells with a T cell receptor (TCR) repertoire. Our preliminary evidence suggests that mice which inherit a second allele have a dramatic decrease in TCR repertoire diversity and lack many unique TCRs present in either We suspect that MHC-heterozygosity in diabetes-resistant mice leads to substantial gaps in the naïve repertoire which reduces the numbers of autoreactive CD4 + T cells, thus contributing to reduction in risk. This proposal is designed to test whether this is so, in mice expressing a restricted TCR and in normal NOD T1D susceptible mice. The proposed study is expected to yield results capable explaining how the most informative genetic risk factors to T1D (MHCII alleles) shape the T cell repertoire may inform future intervention studies in individuals at risk for developing T1D. I propose the use of a new to pair and sequence TCR α /β chains at much larger scales than currently possible, bioinformatic to compare TCR repertoires and the development of retrogenic mice to validate that pathogenic T are selectively absent from mice afforded protection against autoimmune diabetes. All experiments will be at National Jewish Health, a leader in immunological research, under the guidance of my primary and department chair Philippa Marrack, PhD, FRS. Remaining members of my sponsor team are Prof. Kappler at National Jewish Health, and Prof. Victor Greiff at University of Oslo who will support my training in diabetes and bioinformatics respectively. Moreover, I will consult with my thesis and collaborators for support and guidance in performing these studies. My training program will be supplemented by courses, journal clubs and presentations held through the Immunology PhD program University of Colorado, Denver. Overall, the proposed project and training objectives in this F31 award are devised to train me as a resourceful independent graduate student with a strong understanding how to leverage bioinformatics, new RNA sequencing methods and basic science skills to address questions in autoimmunity. MHC-II molecules

项目概要 1 型糖尿病 (T1D) 与人类 MHC-II 等位基因 HLA-DQ2 和 HLA-DQ8 密切相关。 在携带 MHC-II 等位基因 I-Ag7 的小鼠模型中 糖尿病 保护。 MHC-II 糖尿病。 受限制的 主要组织相容性复合体 父母。 TCR 疾病 剧目 的 和 方法 工作流程 细胞 执行的 赞助 约翰 跨学科的 委员会 还 在 应用 的 杰出的 80 – 90% 的雌性小鼠自发产生自身免疫 6个月内。虽然某些会促进疾病易感性，但其他会提供 在人类中，HLA 等位基因 HLA-DQ6 提供保护，而在小鼠中，有几个等位基因 如果与促进疾病的等位基因共表达，则可以从根本上降低自身免疫性疾病的发病率 我们最近研究了两个 MHC 等位基因与一个 MHC 等位基因的遗传如何影响具有 T 细胞受体 (TCR) 库。我们的初步证据表明，遗传了第二个基因的小鼠 等位基因的 TCR 库多样性急剧下降，并且缺乏许多独特的 TCR 我们怀疑，抗糖尿病小鼠中的 MHC 杂合性导致了初始模型中的巨大差距。 减少自身反应性 CD4 + T 细胞数量，从而有助于减少 风险。该提案旨在测试表达受限 TCR 的小鼠是否确实如此 以及正常 NOD T1D 易感小鼠。拟议的研究预计将产生能够 解释信息最丰富的 T1D 遗传风险因素（MHCII 等位基因）如何塑造 T 细胞库 可能为未来针对有患 T1D 风险的个体的干预研究提供信息。我建议使用新的 以比目前可能的生物信息学规模更大的规模对 TCR α /β 链进行配对和测序 比较 TCR 库和逆转录小鼠的发育，以验证致病性 T 小鼠中选择性缺失，从而提供了针对自身免疫性糖尿病的保护作用。所有实验将 在国家犹太健康中心，免疫学研究的领导者，在我的主要指导下 和系主任 Philippa Marrack，博士，FRS。我的赞助团队的其余成员是教授。 国家犹太健康中心的卡普勒和奥斯陆大学的维克多·格雷夫教授将支持我的研究 分别进行糖尿病和生物信息学培训。此外，我会参考我的论文 以及合作者在进行这些研究时提供的支持和指导。我的训练计划将 通过免疫学博士项目举办的课程、期刊俱乐部和演讲作为补充 科罗拉多大学丹佛分校。总体而言，本次 F31 奖项中拟议的项目和培训目标 旨在将我培养成一名足智多谋、具有深刻理解力的独立研究生 如何利用生物信息学、新的 RNA 测序方法和基本科学技能来解决 自身免疫方面的问题。 MHC-II 分子