Role of Grail in maintaining humoral immune tolerance

Grail 在维持体液免疫耐受中的作用

基本信息

批准号：
9089880
负责人：
Roza Insafetdinovna Nurieva
金额：
$ 8万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-15 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9089880
关键词：
Address Agreement Antibodies Antigens Autoantibodies Autoimmune Diseases Autoimmune Process Autoimmunity B-Cell Activation B-Lymphocytes Bone Marrow CD28 gene CD4 Positive T Lymphocytes Cell physiology Cells Cellular biology Chimera organism Clinical Complex Congenic Mice Data Detection Development Disease Down-Regulation Gene Expression Microarray Analysis Gene Targeting Generations Genes Genetic Programming Goals Health Helper-Inducer T-Lymphocyte Humoral Immunities Immune Tolerance Immune response Immunization In Vitro Investigation Knockout Mice Knowledge Lead Lupus Lymphocyte Lymphocyte Function Lymphoid Tissue Maintenance Mediating Methods Mus Pathogenesis Patients Phenotype Play Prevention Production Property Reagent Regulation Reporter Reporting Role Serum Signal Transduction Staging Structure Structure of germinal center of lymph node Symptoms Systemic Lupus Erythematosus T-Cell Development T-Lymphocyte Technology Testing Time Work aging gene anergy anti-dsDNA antibodies autoreactive B cell autoreactive T cell body system central tolerance ds-DNA gene function gene repression insight knock-down lupus prone mice lupus-like mouse model new therapeutic target novel novel therapeutic intervention peripheral tolerance prevent programs research study screening systemic autoimmune disease ubiquitin ligase ubiquitin-protein ligase yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Autoimmunity is initiated by the dysregulation of central or peripheral tolerance, resulting in the escape of autoreactive T and B cells from normal selection. There is rising evidence that expansion of follicular helper T (Tfh) and B cells results in humoral and cellular abnormalities and leads to the development of systemic autoimmune diseases such as Systemic lupus erythematosus (SLE); however, current knowledge of the mechanisms that control autoreactive B and T cell function remains incomplete. Thus, today it is becoming clear that understanding of Tfh and B cell biology and ways to manipulate their function may prove to be invaluable for the treatment of autoantibody-driven diseases. Recently, we generated and analyzed mice deficient in GRAIL and found that aged GRAIL deficient mice developed SLE-like symptoms characterized by massive germinal center formation, accumulation of Tfh and B cells in the lymphoid tissues and high titers of anti-dsDNA antibodies. In addition, GRAIL KO mice crossed with lupus prone B6.Faslpr/lpr mice developed severe lupus phenotype compared to control mice. Moreover, GRAIL expression was down regulated in T and B lymphocytes of patients with active SLE and lupus prone mice, further supporting the role of GRAIL in controlling humoral immune responses. All these findings suggest that GRAIL could be essential in controlling T and B cell functions, and importantly GRAIL down-regulation could serve as a marker for SLE-like disorders. However, it is not clear whether lack of GRAIL in T and/or B cells is responsible for development of lupus-like autoimmunity. We therefore hypothesize that regulation of B and Tfh cell function by GRAIL may be an important checkpoint in humoral tolerance, which is crucial to prevent development of autoimmunity. In Aim1, we will investigate the role of GRAIL in Tfh cell development and function by utilizing gene knockdown approaches. We will employ unique Bcl6-RFP and Bcl6-RFP/Foxp3GFP reporter mouse models to define the function of GRAIL in Tfh and T follicular regulatory (Tfr) cell development in normal and autoimmune settings. In addition, we will identify targets of GRAIL through which it facilitates control of Tfh cell programming by using microarray analysis of gene expression. We have developed a new method of generation of Tfh cells in vitro, which will allow generation of sufficient amount of cells for the proposed analysis. In Aim2, we will assess the role of GRAIL in B cell activation and function. Importantly, we will employ cutting edge technologies, including two-hybrid screening, to identify the exact target(s) of GRAIL that determines its function in B cells. Our results will provide mechanistic insights into the understanding of autoantibody-mediated autoimmune diseases and may lead to identify new therapeutic targets for their detection and prevention.

描述（由适用提供）：自身免疫性是通过中心耐受性或外围耐受性失调而引发的，从而导致自动反应性T和B细胞从正常选择中逃脱。有越来越多的证据表明卵泡辅助剂T（TFH）和B细胞的膨胀结果在体液和细胞异常中，导致系统性自身免疫性疾病（如全身性红斑狼疮）（SLE）的发展；但是，当前对控制自动反应性B和T细胞功能的机制的了解仍然不完整。这就是如此，今天很清楚，对TFH和B细胞生物学的理解以及操纵其功能的方法可能被证明对治疗自身抗体驱动的疾病是无价的。最近，我们产生并分析了缺乏圣杯的小鼠，发现陈年的小鼠缺乏小鼠出现了类似SLE的症状，其特征是以大量生发中心形成，淋巴组织中TFH和B细胞的积累以及抗DSDNA抗体的高滴度。此外，与对照小鼠相比，与狼疮的b6.faslpr/LPR小鼠交叉的Grail Ko小鼠相比，发生了严重的狼疮表型。此外，在活跃的SLE和狼疮小鼠的患者的T和B淋巴细胞中调节了圣杯表达，进一步支持了圣杯在控制体液免疫调查中的作用。所有这些发现表明，圣杯对于控制T和B细胞功能至关重要，重要的是，Grail下调可能是SLE样疾病的标志物。但是，目前尚不清楚T和/或B细胞中缺少Grail是否导致狼疮样自身免疫性的发展。因此，我们假设通过圣杯对B和TFH细胞功能的调节可能是体液耐受性的重要检查点，这对于防止自身免疫的发展至关重要。在AIM1中，我们将通过利用基因敲低方法来研究Grail在TFH细胞开发和功能中的作用。我们将采用唯一的BCL6-RFP和BCL6-RFP/FOXP3GFP记者鼠标模型来定义正常的TFH和T卵泡调节（TFR）细胞开发中Grail的功能和自动免疫设置。此外，我们将通过使用基因表达的微阵列分析来确定它通过该目标托管TFH细胞编程的控制目标。我们已经开发了一种在体外生成TFH细胞的新方法，该方法将允许生成足够数量的细胞进行拟议的分析。在AIM2中，我们将评估Grail在B细胞激活和功能中的作用。重要的是，我们将采用尖端技术，包括两种杂交筛选，以确定确定其在B细胞中功能的手指的确切目标。我们的结果将提供对自身抗体介导的自身免疫性疾病的理解的机械见解，并可能导致确定其检测和预防的新治疗靶标。