Influence of ligand on specification of gamma/delta fate and function

这个提议的目标是深入了解控制yS谱系定型的分子过程 和效应子命运的特异性。yS谱系定型和效应子命运的特化均发生 然而，我们对控制胸腺发育的发育线索的理解， 这些命运决定仍然不完整。越来越多的证据表明， T细胞受体（TCR）信号强度的差异，通过E盒DNA的分级抑制表现出来 结合蛋白（E蛋白）介导的E蛋白拮抗剂，IdS。然而，E蛋白靶向 对这些命运决定至关重要的因素仍然没有明确的定义。目前还不清楚不同的Y 5是否 TCR复合物与交替命运选择相关，通过自主转导 不同的强度或是否需要配体接合。为了解决这些问题，我们将利用 理想的合适的ySTCR转基因模型（KN 6），其已知的选择配体，非经典MHC-I T-10/22型微处理器，可用于改变所得TCR信号。在艾米，我们将：采用KN 6 tg 小鼠以及内源性T-10/22反应性yS祖细胞，以确定如何特异性消融T-10/22反应性yS祖细胞。 10/22配体影响γ S谱系定型、库选择和效应子功能。AIM 2旨在 理解IdS是VY 2+发育所需的矛盾观察的基础， VyS-t- yS T细胞，但抑制Vyi.1+先天性yS T细胞的发育。我们将评估 在不存在IdS的情况下Vy1.1+先天性γ S T细胞的扩增是Vy1.1A/66.3的自主属性。 TCR复合物或需要配体接合。AimS解决了关键的未解决的问题， y 6谱系定型和效应子命运的指定是可分离的或同时发生的。为此， 我们将利用我们新描述的γ S谱系定型的标志物，CD 7S诱导。全基因组ChlP- 还将在CD 73标记的细胞上进行E蛋白靶点的Seq，以组装全局调控序列。 网络定义的承诺过程。这些努力需要联合所有国家的能力， 这个项目的成员，承诺揭示关键的新的见解如何γ S T细胞的发展是 控制。 相关性（参见说明）： Y 5 T细胞调节炎症，保护上皮屏障，特别擅长杀死皮肤 肿瘤的因此，了解控制其发育和功能的分子过程 可以使它们的操作具有治疗益处。此外，我们对分子效应器的研究 控制T细胞系定型对其它发育过程也是至关重要的， 因为细胞生长和分化的控制是发育和转化中反复出现的主题。

The goal of this proposal is to gain insight into the molecular processes controlling yS lineage commitment and specification of effector fate. Both yS lineage commitment and specification of effector fate occur during development in the thymus; however, our understanding of the developmental cues controlling these fate decisions remains incomplete. Accumulating evidence suggests that they are governed by differences in T cell receptor (TCR) signal strength that manifest through graded repression of E box DNA binding proteins (E proteins) mediated by the E protein antagonist, IdS. Nevertheless, the E protein targets that are crucial for these fate decisions remain poorly defined. It is also unclear whether the different Y5 TCR complexes linked to alternate fate choices promote them by autonomously transducing signals of differing intensities or if they require ligand-engagement. In addressing these questions, we will exploit an ideally suited ySTCR transgenic model (KN6) whose known selecting ligand, the non-classical MHC-I nnoleculeT-10/22,CanbemanipulatedtoaltertheresultantTCRsignal.InAimi,wewill:employKN6tg mice as well as endogenous T-10/22 reactive yS progenitors to determine how specific ablation of the T- 10/22 ligand affects yS lineage commitment, repertoire selection, and effector function. Aim2 seeks to understand the basis for the paradoxical observation that IdS is required for the development of VY2+ and VyS-t- yS T cells, but restrains the development of Vyi.1+ innate yS T cells. We will assess whether the expansion of Vyi.1+ innate yS T cells in the absence of IdS is an autonomous attribute of the Vyl.l A/66.3 TCR complex or requires ligand-engagement. AimS addresses the critical unresolved question of whether y6 lineage commitment and specification of effector fate are separable or occur simultaneously. To do so, we will utilize our newly described marker of yS lineage commitment, CD7S induction. Genome wide ChlP- Seq on E protein targets will also be performed on CD73-marked cells to assemble a global regulatory network defining the commitment process. These efforts, which require the combined capabilities of all of the members of this program, promise to reveal critical new insights into how yS T cell development is controlled. RELEVANCE (See instructions): Y5 T cells regulate inflammation, preserve epithelial barriers, and are particularty adept at killing cutaneous tumors. Accordingly, understanding the molecular processes controlling their development and function may enable their manipulation for therapeutic benefit. Moreover, our investigation of molecular effectors controlling T lineage commitment is also of fundamental importance for other developmental processes, since control of cell growth and differentiation is a recurring theme in development and transformation.