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: 6891598
• 负责人: Laurent Brossay
• 金额: $ 30.52万
• 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/6891598
• 关键词: Betaherpesvirinae; Herpesviridae disease; T lymphocyte; cell growth regulation; cell morphology; expression cloning; immunologic receptors; laboratory mouse; lectin; leukocyte activation /transformation; natural killer cells; receptor expression; reporter genes; tissue /cell culture; transfection

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细胞在针对病原体（如鼠巨细胞病毒（MCMV））的先天防御中发挥关键作用。NK细胞表面上存在的活化性和抑制性受体与先天性细胞因子协同作用以调节NK细胞功能。最近，我们发现杀伤细胞凝集素样受体G1（KLRG 1）表达于最成熟的NK细胞上，MCMV感染诱导NK细胞和CD 8 + T细胞表面KLRG 1分子的上调。我们还证明了KLRG 1接合可以抑制NK细胞效应子功能。此外，我们产生了KLRG 1四聚体，并通过流式细胞术鉴定了四聚体阳性的细胞系。基于这些初步的数据，我们因此建议研究KLRG 1功能的几个方面。在特定目标1中，我们将使用KLRG 1作为成熟NK细胞的标志物，以研究NK细胞对MCMV感染的反应性房室扩增和收缩阶段。我们还将检查在MCMV感染小鼠的情况下体内阻断KLRG 1功能的后果，以确定KLRG 1功能是否是防止NK细胞和/或CD 8 v T细胞的不受控制的活化。在具体目标2中，我们将研究决定KLRG 1分子抑制特性的生化机制。为此，将进行野生型和突变型KLRG 1分子的转染/表达分析。在特定目标3中，我们将使用报告细胞系方法，以确认我们的四聚体染色的特异性并鉴定表达KLRG 1配体的细胞。在特定目标4中，将开发表达克隆策略以克隆KLRG 1配体。这些新的研究将为NK细胞和CD 8 + T细胞上KLRG 1的特异性和免疫调节作用以及NK受体的一般作用提供见解，并为调节这一重要类别的免疫细胞应答提供策略。