Antigen Discovery in Acute Kidney Injury

急性肾损伤中抗原的发现

• 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
• 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

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(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有显著的不利影响。我们小组和其他人在肾、肝、肺、肠和脑中的最新数据表明，淋巴细胞，特别是CD4T细胞，在IRI中起着直接作用。T细胞通常对同种异体抗原或修饰的自身抗原有反应。到目前为止，IRI中引发炎症和组织损伤的抗原尚不清楚。主要申请者将与经验丰富的蛋白质化学家和基础免疫学家合作，为肾脏IRI领域的高风险、高影响、新的研究路线提供跨学科的协同作用。假设：在IRI中，自身抗原在驱动T细胞的炎症和免疫反应中起重要作用。具体目的1.建立小鼠肾夹夹致温热性IRI模型。我们将从IRI和Sham动物身上获取肾周淋巴结，并使用人类白细胞抗原II类免疫沉淀，分离IRI提呈的多肽。利用高度复杂的蛋白质分离和鉴定技术，包括MHC-肽复合体的高效液相色谱/质谱分析，我们将鉴定参与肾脏IRI期间T细胞免疫反应的蛋白质的特定表位。目的2.将合成的合成肽T细胞表位注射给幼鼠，并在选择的时间对IRI的功能、组织学、分子和免疫学参数进行评估。我们将在体外将分离的合成肽负载到树突状细胞(DC)，然后将负载的DC注射到小鼠体内。在注射多肽或负载DC后，小鼠将接受IRI，以确定保护水平和组织反应。预期结果：寻找具有治疗潜力的新多肽，以减少IRI造成的组织损伤。 公共卫生相关性：在肾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、免疫学和活体模型相结合的方法来鉴定肾脏中与肾脏IRI相关的多肽。对这些的研究可以导致新的治疗方法，以减少肾IRI后的组织损伤和炎症。