Defining the SAP-dependent and SAP-independent gamma delta TCR repertoire

定义 SAP 相关和 SAP 独立的 gamma delta TCR 指令集

基本信息

批准号：
10170255
负责人：
JONATHAN E BOYSON
金额：
$ 7.8万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-22 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10170255
关键词：
Adaptor Signaling Protein Adult Affect Applications Grants Area Bacteria Bar Codes Birth Cells Characteristics Data Data Set Dependence Development Embryo Exhibits Family Frequencies Hematopoietic Immune response Impairment Interferons Interleukin-17 Interleukin-4 Investigation Laboratories Libraries Lung Lymphocyte Lymphoid Tissue Microbe Mucous Membrane Mus Organ Pathway interactions Peripheral Play Population Receptor Signaling Research Personnel Resolution Role SH2D1A gene SLAM family receptor Shapes Signal Pathway Signal Transduction Skin Spleen T-Cell Development T-Cell Receptor T-Lymphocyte T-Lymphocyte Subsets T-cell receptor repertoire Thymus Gland Tissues Tumor Immunity Virus adaptive immune response anti-tumor immune response antimicrobial chemokine cytokine fungus neonate next generation next generation sequencing novel receptor

项目摘要

The tissue-resident innate-like T cell subsets (i.e., NKT, MAIT, and innate-like T) constitute major lymphocyte populations in barrier tissues like the lung, gut, and skin, where they provide a critical first line of defense against bacteria, fungi, and viruses and where they are active players in anti-tumor immunity. Innate- like T cell subsets all share the ability to rapidly producing large amounts of cytokines and chemokines, and thereby possess the ability to shape both the quantity and quality of both the innate and adaptive immune responses. Although the NKT and mucosal-associated invariant T (MAIT) innate-like  T cell subsets are well-studied, far less is known of the innate-like  T cell populations, and their identity remains ill-defined. We recently completed a comprehensive investigation into the role of the SLAM/SAP signaling pathway in  T cell development and function. This study revealed that the SLAM/SAP signaling pathway is a critical determinant in the thymic development of both IL-17- and IFN--producing  T cells. Since one of the hallmarks of nearly all innate-like T cell subsets is a dependence on the SLAM/SAP signaling pathway for their development, these results raise the possibility that SAP regulates the development of, as yet undefined innate-like  T cell subsets. Therefore, the objective of this proposal is to unambiguously define the SAP-dependent  T cell subsets. Because a characteristic feature of all innate-like T cells identified so far is the expression of a signature invariant of semi-invariant TCR, we propose to: identify the SAP-dependent and SAP-independent  TCR clonotypes using single-cell next-generation sequencing. We will sort single  T cells from C57BL/6J and B6.SAP-/- mice, and determine the paired TCR clonotypes by sequencing barcoded amplicon libraries generated using V- and V-specific primers. We will examine thymic  T cells at different stages of development (embryonic day 17, birth, day 6 neonate, and adult) to capture distinct waves of  T cell development, and we will examine lung and spleen to identify the SAP-dependent subsets in the periphery. Upon completion of these studies, we will have unambiguously identified the SAP-dependent  TCR clonotypes, which will be critical in to our understanding how SLAM/SAP signaling regulates the developmental programming of  T cells. In addition, we will have provided an invaluable high-resolution dataset of the paired  TCR clonotypes in both the thymus and periphery to investigators seeking to understand better the role of  T cells in the immune response.

组织居住的先天性T细胞子集（即NKT，MAIT和HINATE-liket）构成主要肺，肠道和皮肤等屏障组织中的淋巴细胞种群，它们提供关键的第一线防御细菌，真菌和病毒，以及他们在抗肿瘤免疫中活跃的参与者。先天- 像T细胞子集一样从而具有塑造先天性和适应性免疫的数量和质量的能力响应。尽管NKT和与粘膜相关的不变t（Mait）先天性t细胞子集是经过充分研究，众所周知的是先天性的t细胞群体，其身份仍然不确定。我们最近完成了对th细胞中SLAM/SAP信号通路的作用的全面研究发展和功能。这项研究表明，猛击/SAP信号通路是关键的确定器在IL-17-和IFN-产生T细胞的胸腺发育中。由于几乎是所有先天的T细胞子集都依赖于猛击/SAP信号传导途径的发育，这些结果增加了SAP调节发展的可能性子集。因此，该提议的目的是明确定义SAP依赖性T细胞子集。因为到目前为止确定的所有先天性T细胞的特征是A的表达半不变TCR的签名不变，我们提出：确定依赖SAP的和SAP独立的 使用单细胞下一代测序的TCR克隆型。我们将从C57BL/6J和 b6.sap - / - 小鼠，并通过对条形码的扩增子库来确定配对的TCR clonotypes 使用V-和V特异性引物生成。我们将在不同阶段的不同阶段检查胸腺T细胞开发（胚胎第17天，出生，第6天的新生儿和成人）以捕获不同的T细胞的独特波发展，我们将检查肺和脾脏，以识别周围依赖SAP的子集。完成这些研究后，我们将明确识别SAP依赖性TCR clonotypes，这对于我们了解如何猛击/SAP信号调节发育至关重要 T细胞的编程。此外，我们还将提供配对的宝贵的高分辨率数据集 tcr的胸腺和外围的TCR clonotypes in to研究者寻求更好地理解的作用免疫响应中的T细胞。