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

基本信息

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
项目类别：
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

项目摘要

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

项目摘要 1型糖尿病（T1D）与人类MHC-II等位基因HLA-DQ2和HLA-DQ8密切相关。在带有MHC-II等位基因I-AG7的鼠标中糖尿病保护。 MHC-II 糖尿病。受限制的 MHC 父母。 TCR 疾病曲目的和方法工作流程细胞执行赞助约翰跨学科委员会还在应用的杰出的 80 - 90％的雌性小鼠发育于自身免疫性在6个月内。虽然某些促进疾病的敏感性，但其他人则提供在人类中，HLA等位基因HLA-DQ6提供了保护，而在老鼠中有几个等位基因如果与促进等位基因的疾病共表达，从根本上降低了自身免疫的发生率我们最近研究了两个MHC等位基因与一个遗传如何影响T细胞 T细胞受体（TCR）曲目。我们的初步证据表明，继承第二个的小鼠等位基因的TCR曲目多样性急剧下降，并且在任何一个中都缺乏许多独特的TCR 我们怀疑抗糖尿病的小鼠中的MHC杂合性导致幼稚的差距很大减少自动反应性CD4 + T细胞数量的曲目，从而有助于减少风险。该建议旨在测试是否是这样的，在表达有限TCR的小鼠中在正常的点头T1D易感小鼠中。拟议的研究预计将产生能力的结果解释T1D（MHCII等位基因）最有用的遗传危险因素如何塑造T细胞库可能会为未来的干预研究提供给有风险开发T1D的个体的干预研究。我建议使用新的在比当前可能更大的尺度上配对和序列TCRα /β链比较TCR曲目和后源性小鼠的发展以验证该致病性T 从小鼠中选择性地没有自身免疫性糖尿病的保护。所有实验将是在我的主要的指导下和部门主席Philippa Marrack博士，FRS。我的赞助商团队的其余成员是教授 National Jewish Health的Kappler和奥斯陆大学的Victor Greiff教授将支持我分别接受糖尿病和生物信息学的培训。而且，我将咨询我的论文以及协作者在进行这些研究方面的支持和指导。我的培训计划将通过免疫学博士学位计划进行的课程，期刊俱乐部和演讲补充科罗拉多大学丹佛大学。总体而言，该F31奖中的拟议项目和培训对象被设计为训练我是一名足智多谋的独立研究生如何利用生物信息学，新的RNA测序方法和基础科学技能来解决自身免疫性问题。 MHC-II分子