Antigen Discovery in Acute Kidney Injury

急性肾损伤中抗原的发现

• 批准号: 8107544
• 负责人: HAMID RABB
• 金额: $ 20.02万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 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/8107544
• 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; High Pressure Liquid Chromatography; Histocompatibility Antigens Class II; Histologic; Immune; Immune response; Immunity; 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

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缺血再灌注损伤（IRI）中驱动炎症反应的抗原。肾脏 IRI 对自体肾脏和同种异体移植肾具有显着的有害影响。我们小组和其他人在肾脏、肝脏、肺、肠和大脑中的最新数据表明淋巴细胞，特别是 CD4 T 细胞，在 IRI 中具有直接作用。 T 细胞通常对同种异体抗原或修饰的自身抗原作出反应。迄今为止，IRI 中引发炎症和组织损伤的抗原尚不清楚。主申请人将与经验丰富的蛋白质化学家和基础免疫学家合作，跨学科协同开展肾脏 IRI 的高风险、高影响、新颖的研究。假设：自身抗原在 IRI 中驱动 T 细胞炎症和免疫反应中发挥重要作用。具体目标 1. 我们将使用已建立的肾钳诱导温热 IRI 小鼠模型。我们将从IRI和假手术动物中获取肾周围淋巴结，并使用HLA-II类免疫沉淀，分离IRI呈递的肽。使用高度复杂的蛋白质分离和鉴定技术，包括 MHC-肽复合物的 HPLC/质谱分析，我们将鉴定肾 IRI 期间参与 T 细胞免疫反应的蛋白质的特定表位。目标 2. 我们将目标 1 中确定的合成肽 T 细胞表位注射到初始小鼠中，并在选定的时间评估 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加载到树突状细胞（DC）中，然后将这些加载的DC注射到小鼠体内。注射肽或负载 DC 后，将小鼠接受 IRI 以确定保护水平和组织反应。预期结果：找到具有治疗潜力的新型肽，以减少 IRI 造成的组织损伤。 公共卫生相关性：在肾脏缺血再灌注损伤中驱动炎症反应的抗原尚不清楚。我们计划使用蛋白质化学、免疫学和体内模型相结合的方法来鉴定肾脏中参与肾脏 IRI 的肽。这些研究可以带来新的治疗方法，以减少肾脏 IRI 后的组织损伤和炎症。