Characterization of the polyclonal TCR repertoire in TCR-beta transgenic mice

描述(由申请人提供)：随机生成的T细胞受体(TCR)序列确保了TCR谱系能够识别各种病原体。然而，这种巨大的多样性阻碍了对TCR曲目的研究。为了解决这个问题，我们使用了一个固定的TCR-β链模型来促进对多克隆TCR谱系的实验研究。这种TCR谱系在正常的胸腺和外周环境中发育，并受到自然T细胞选择性的影响，包括与皮肤和肠道共生菌的自身抗原和外来抗原的潜在相互作用。在以前的研究中，这种固定的TCR-β链模型在描述正常的自然产生的TCR谱系以及将它们与自身免疫谱系进行比较方面是有用的。我们在这里提出，在一个固定的TCR-β链模型中详细了解TCR谱系将有助于T细胞生物学的许多方面的研究，因此，本项目的目标是通过对基于空间和表型变量分离的T细胞中可变的TCR-α链进行广泛的测序，在单个TCR水平上完整地原位表征这个多克隆TCR谱系。一个特别的重点领域将是了解自然产生的记忆CD4T细胞的TCR特异性，这些细胞可能是由于遇到自身或共生抗原而产生的，并可能是潜在的致病细胞。我们相信，这些数据将成为未来研究自我反应性调节性和非调节性T细胞、自身免疫性疾病的遗传模型、记忆性T细胞以及T细胞运输和动态平衡的基础。

DESCRIPTION (provided by applicant): The random generation of T cell receptor (TCR) sequences ensures a TCR repertoire capable of recognizing a wide variety of pathogens. However, study of the TCR repertoire is hampered by this great diversity. To overcome this issue, we have used a fixed TCR-beta chain model to facilitate experimental study of a polyclonal TCR repertoire. This TCR repertoire develops in a normal thymic and peripheral environment, and is subject to natural T cell selective forces, including potential interactions with self-antigens and foreign antigens from commensal flora on the skin and intestines. In previous studies, this fixed TCR-beta chain model has been useful in characterizing normal naturally arising TCR repertoires and comparing them with autoimmune repertoires. We propose here that detailed knowledge of the TCR repertoire in a fixed TCR- beta chain model will be useful for study of numerous aspects of T cell biology, Thus, the goal of this project is the complete in situ characterization of this polyclonal TCR repertoire at the individual TCR level by extensive sequencing of the variable TCR-alpha chains from T cells isolated based on spatial and phenotypic variables. One particular area of focus will be to understand the TCR specificity of naturally arising memory CD4+ T cells, which presumably arose due to encounter with self or commensal antigens, and may potentially be pathogenic. We believe these data will serve as the foundation of future studies on self-reactive regulatory and non-regulatory T cells, genetic models of autoimmune disease, memory T cells, and T cell trafficking and homeostasis.