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

项目摘要 1型糖尿病（T1 D）与人类MHC-II等位基因HLA-DQ 2和HLA-DQ 8密切相关。 在携带MHC-II等位基因I-Ag 7的小鼠模型中， 糖尿病 保护 MHC-II 糖尿病 限制 MHC 家长 TCR 疾病 汇辑 的 和 方法 工作流 细胞 执行 申办者 约翰 跨学科 委员会 也 在 应用 的 优秀 80 - 90%的雌性小鼠会自发发生自身免疫性疾病 在6个月内虽然某些促进疾病的易感性，其他提供 在人类中，HLA等位基因HLA-DQ 6提供保护，而在小鼠中，有几个等位基因HLA-DQ 6提供保护。 其如果与疾病促进等位基因共表达，则从根本上降低自身免疫性疾病的发生率。 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）库。我们的初步证据表明， 等位基因的TCR库多样性显著降低，并且缺乏许多存在于任一等位基因中的独特TCR。 我们怀疑，糖尿病抗性小鼠的MHC杂合性导致了幼稚细胞中的大量缺口。 减少自身反应性CD 4 + T细胞的数量，从而有助于减少 风险这项建议旨在测试是否如此，在小鼠中表达限制性TCR 和正常NOD T1 D易感小鼠。预计拟议的研究将产生能够 解释T1 D（MHCII等位基因）最具信息性的遗传风险因素如何塑造T细胞库 可能会为未来在有发展T1 D风险的个体中进行的干预研究提供信息。我建议使用一种新的 以比目前可能的更大的规模配对和测序TCR α /β链，生物信息学 比较TCR库和逆转录小鼠的发育，以验证致病性T细胞 被选择性地从提供针对自身免疫性糖尿病的保护的小鼠中缺失。所有的实验都将 在国家犹太健康，免疫学研究的领导者，在我的主要指导下， 和部门主席菲利帕马拉克，博士，FRS。我的申办者团队的其余成员是Prof. 国家犹太人健康中心的Kappler和奥斯陆大学的维克托格列夫教授将支持我的研究。 分别接受糖尿病和生物信息学培训。此外，我将参考我的论文， 和合作者在进行这些研究的支持和指导。我的训练计划将 通过课程，期刊俱乐部和通过免疫学博士课程举行的演讲来补充 丹佛市科罗拉多大学。总的来说，F31项目的建议和培训目标 旨在培养我成为一个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 关于自身免疫的问题 MHC-II分子

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