Functions of the KLRG1 molecule on NK cells and T cells

KLRG1分子对NK细胞和T细胞的功能

• 批准号: 7221914
• 负责人: Laurent Brossay
• 金额: $ 28.92万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7221914
• 关键词: Binding; Biochemical; Biological; CD8-Positive T-Lymphocytes; CD8B1 gene; Categories; Cell Line; Cell physiology; Cell surface; Cells; Cytomegalovirus; Cytomegalovirus Infections; Data; Effector Cell; Flow Cytometry; Histocompatibility Antigens Class I; ITIM; Immune; Infection; Killer Cells; Lectin; Ligands; Murid herpesvirus 1; Mus; Natural Killer Cells; Phase; Play; Property; Regulation; Reporter; Research Personnel; Role; Signal Transduction; Specificity; Staining method; Stains; Surface; T-Lymphocyte; Transcriptional Activation; Transfection; Tyrosine; Up-Regulation; Virus Diseases; base; cytokine; design; expression cloning; in vivo; insight; mutant; novel; pathogen; prevent; programs; receptor; research study; response; trafficking

DESCRIPTION (provided by applicant): NK cells play a crucial role in innate defense against pathogens such as murine cytomegalovirus (MCMV). The presence of both activating and inhibitory receptors on the surface of NK cells act collaboratively and in concert with innate cytokines to regulate NK cell functions. Recently, we showed that the killer cell lectin-like receptor G 1 (KLRG 1) is expressed on the most mature NK cells and that infection with MCMV induces an up-regulation of the KLRG1 molecule on the surface of both NK cells and CD8+ T cells. We also demonstrated that KLRG 1 engagement can inhibit NK cell effector functions. In addition, we generated the KLRG1 tetramer and have identified cell lines that are tetramer positive by flow cytometry. Based on these preliminary data, we therefore propose to study several aspects of KLRG 1 functions. In Specific Aim 1, we will use KLRG 1 as a marker of mature NK cells to investigate the NK cell compartmental expansion and contraction phases in response to MCMV infection. We will also examine the consequences of the in vivo blocking of KLRG 1 functions in the context of the MCMV infection of mice in order to determine whether KLRG1 function is to prevent the uncontrolled activation of both NK cells and/or CD8 v T cells. In Specific Aim 2, we will study the biochemical mechanism that dictates the inhibitory properties of the KLRG1 molecule. To do this, transfection/expression analyses of wild-type and mutant KLRG1 molecules will be performed. In Specific Aim 3, we will use a reporter cell line approach, to both confirm the specificity of our tetramer staining and identify cells that express the KLRG1 ligand. In Specific Aim 4, an expression cloning strategy will be developed to clone the KLRG1 ligand. These novel studies will provide insights into the specificity and immunoregulatory role of KLRG1 in particular and NK receptors in general on both NK cells and CD8+ T cells, as well as provide strategies for the regulation of this important category of immune cell responses.

描述（由申请人提供）：NK 细胞在针对鼠巨细胞病毒（MCMV）等病原体的先天防御中发挥着至关重要的作用。 NK 细胞表面存在激活性和抑制性受体，与先天细胞因子协同作用，调节 NK 细胞功能。最近，我们发现杀伤细胞凝集素样受体 G 1 (KLRG 1) 在最成熟的 NK 细胞上表达，MC​​MV 感染会诱导 NK 细胞和 CD8+ T 细胞表面的 KLRG1 分子上调。我们还证明 KLRG 1 的参与可以抑制 NK 细胞效应功能。此外，我们生成了 KLRG1 四聚体，并通过流式细胞术鉴定了四聚体阳性的细胞系。因此，基于这些初步数据，我们建议研究 KLRG 1 功能的几个方面。在具体目标 1 中，我们将使用 KLRG 1 作为成熟 NK 细胞的标记物来研究 NK 细胞区室扩张和收缩阶段对 MCMV 感染的反应。我们还将检查在小鼠 MCMV 感染的情况下体内阻断 KLRG 1 功能的后果，以确定 KLRG1 功能是否可以防止 NK 细胞和/或 CD8 v T 细胞不受控制的激活。在具体目标 2 中，我们将研究决定 KLRG1 分子抑制特性的生化机制。为此，将对野生型和突变型 KLRG1 分子进行转染/表达分析。在特定目标 3 中，我们将使用报告细胞系方法来确认四聚体染色的特异性并鉴定表达 KLRG1 配体的细胞。在具体目标 4 中，将开发表达克隆策略来克隆 KLRG1 配体。这些新研究将深入了解 KLRG1（特别是 KLRG1）和 NK 受体（一般）对 NK 细胞和 CD8+ T 细胞的特异性和免疫调节作用，并为调节这一重要类别的免疫细胞反应提供策略。