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: 10468670
• 负责人: Alexander J Brown
• 金额: $ 3.47万
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468670
• 关键词: Address; Affect; Alleles; Animals; Antibodies; Antigens; Autoimmune; Autoimmune Diabetes; Autoimmune Diseases; Autoimmunity; Award; B-Cell Antigen Receptor; Bar Codes; Basic Science; Beta Cell; 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; 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; 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

项目总结 1型糖尿病(T1D)与人类MHC-II等位基因HLADQ2和HLADQ8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 MHC 家长。 TCR 疾病 汇辑 的 和 方法 工作流程 单元格 已执行 赞助商 约翰 跨学科 委员会 也是 在… 应用程序 的 杰出的 80%-90%的雌性小鼠自发产生自身免疫 在6个月内。虽然某些促进疾病易感性，但其他人提供 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，可以从根本上降低自身免疫的发生率 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T细胞 T细胞受体(TCR)谱系。我们的初步证据表明，遗传第二次的小鼠 等位基因在TCR谱带多样性中显著降低，并且在这两个基因中都缺乏许多独特的TCR 我们怀疑糖尿病抵抗小鼠的MHC杂合性导致了幼稚的巨大差距 减少自身反应的CD4+T细胞的数量，从而有助于减少 风险。这项建议是为了在表达限制性TCR的小鼠身上测试是否如此 正常NOD T1D易感小鼠。预计拟议的研究将产生有能力的结果 解释T1D(MHCII等位基因)的最具信息量的遗传风险因素如何塑造T细胞谱系 可能会为未来针对T1D高危人群的干预研究提供参考。我建议使用一种新的 在比目前可能的规模更大的范围内配对和测序tcrα/β链，生物信息学 比较TCR谱系和逆转录基因小鼠的发育情况，以验证致病T细胞 在对自身免疫性糖尿病提供保护的小鼠中选择性地缺失。所有的实验都将是 在国家犹太人健康中心，免疫学研究的领先者，在我的初级指导下 系主任菲利帕·马拉克，博士，FRS。我的赞助商团队的其余成员是。 国家犹太人健康中心的Kappler和奥斯陆大学的Victor Greiff教授将支持我的 分别在糖尿病和生物信息学方面进行培训。此外，我会参考我的论文 和合作者在执行这些研究时提供支持和指导。我的培训计划将 辅以通过免疫学博士项目举办的课程、杂志俱乐部和演讲 科罗拉多大学丹佛分校。总体而言，该F31奖项中建议的项目和培训目标 是为了把我培养成一个足智多谋、理解力强的独立研究生 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 关于自身免疫力的问题。 MHC-II分子