Antigen Discovery in Acute Kidney Injury

急性肾损伤中抗原的发现

基本信息

批准号：
7989727
负责人：
HAMID RABB
金额：
$ 24.26万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-15 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7989727
关键词：
Acute Renal Failure with Renal Papillary Necrosis Alloantigen Allografting Animals Antigens Autoantigens Automobile Driving Brain CD4 Positive T Lymphocytes Cations Data Dendritic Cells Development Discipline Disease Disease model Dose Epitopes Generations Goals Histocompatibility Antigens Class II Histologic Immune Immune response Immunologist Immunology Immunoprecipitation In Vitro Inflammation Inflammatory Inflammatory Response Injury Intestines Investigation Ischemic Bowel Disease Kidney Lead Leukocytes Liver Long-Term Effects Lung Lymphocyte MHC Class II Genes Mass Spectrum Analysis Mediating Molecular Mononuclear Leukocytes Mus Organ Outcome Pathogenesis Peptide T Peptide/MHC Complex Peptides Play Process Protein Chemistry Proteins Renal function Reperfusion Injury Research Role Route T cell response T-Cell Depletion T-Lymphocyte T-Lymphocyte Epitopes Techniques Technology Testing Therapeutic Time Tissues Warm Ischemia Work delayed graft function experience high risk human disease improved in vivo Model innovation interdisciplinary approach interest kidney allograft lymph nodes member mouse model neutrophil novel novel strategies novel therapeutics protein aminoacid sequence public health relevance renal ischemia response reversed phase chromatography synthetic peptide

项目摘要

DESCRIPTION (provided by applicant): The objective of this application is to elucidate the antigens that drive the inflammatory response in kidney ischemia reperfusion injury (IRI). Kidney IRI has a significant detrimental effect on native kidneys and allografts. Recent data from our group and others in kidney, liver, lung, intestine and brain has shown a direct role for lymphocytes, particularly CD4 T cells, in IRI. T cells typically respond to alloantigens, or modified self antigens. To date, the antigens that fuel inflammation and tissue injury in IRI are not known. The primary applicant will partner with an experienced protein chemist and basic immunologists to synergize across disciplines for a high risk, high impact, novel line of investigation in kidney IRI. Hypothesis: Self antigens play an important role in driving inflammation and immune responses of T cells in IRI. Specific Aim 1. We will use an established mouse model of kidney clamp induced warm IRI. We will obtain peri-renal lymph nodes from IRI and sham animals, and use HLA-class II immunoprecipitation, isolate the IRI presented peptides. Using highly sophisticated protein isolation and identification techniques including HPLC/Mass Spectroscopy analysis of MHC-peptide complex, we will identify the specific epitopes of proteins involved in T cell immune responses during renal IRI. Aim 2. We will inject the synthetic peptide T cell epitopes identified in Aim 1 to naive mice, and at select times, the functional, histologic, molecular and immunological parameters of IRI will be evaluated. We will load isolated synthetic peptides to dendritic cells (DCs) in vitro and then these loaded DCs will be injected into mice. After injecting peptides or loaded DC's the mice will be submitted to IRI to determinate the level of protection and tissue response. Expected results: to find novel peptides that will have therapeutic potential to decrease tissue injury from IRI. PUBLIC HEALTH RELEVANCE: The antigens that drive the inflammatory responses in kidney ischemia reperfusion injury are unknown. We plan to use a combined protein chemistry, immunology and in vivo model approach to identify the peptides in the kidney that are involved in kidney IRI. The study of these can lead to new therapeutics to decrease tissue injury and inflammation after kidney IRI.

描述（由申请人提供）：本申请的目的是阐明在肾脏缺血再灌注损伤（IRI）中驱动炎症反应的抗原。肾脏IRI对本地肾脏和同种异体移植物具有重大不利影响。在IRI中，来自肾脏，肝，肺，肠和大脑的肾脏，肝，肺，肠和大脑的最新数据表现出了淋巴细胞，尤其是CD4 T细胞的直接作用。 T细胞通常对同种抗原或修饰的自抗原反应。迄今为止，尚不清楚IRI燃料炎症和组织损伤的抗原。主要的申请人将与经验丰富的蛋白质化学家和基本免疫学家合作，以在肾脏IRI中具有高风险，高影响力，新颖的研究系列，以协同跨学科协同作用。假设：自抗原在驱动IRI中T细胞的炎症和免疫反应中起重要作用。具体目标1。我们将使用已建立的肾脏夹诱导温暖IRI的小鼠模型。我们将从IRI和假动物中获得肾脏周围淋巴结，并使用HLA级II免疫沉淀分离出IRI呈现的肽。使用高度复杂的蛋白质分离和鉴定技术，包括对MHC肽复合物的HPLC/质谱分析分析，我们将确定肾脏IRI期间参与T细胞免疫反应的蛋白质的特定表位。 AIM 2。我们将注入AIM 1中鉴定到幼稚小鼠中鉴定的合成肽T细胞表位，并且将评估IRI的功能，组织学，分子和免疫学参数。我们将在体外将分离的合成肽载至树突状细胞（DC），然后将这些载荷DC注入小鼠。注射肽或加载DC后，将提交给IRI以确定保护水平和组织反应水平。预期结果：找到具有治疗潜力的新型肽，以减少IRI的组织损伤。公共卫生相关性：推动肾脏缺血再灌注损伤中炎症反应的抗原尚不清楚。我们计划使用合并的蛋白质化学，免疫学和体内模型方法来鉴定肾脏IRI涉及的肾脏中的肽。这些研究可能导致新的治疗剂减少肾脏IRI后的组织损伤和炎症。