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驱动肾缺血再灌注损伤（IRI）炎症反应的抗原。肾脏IRI对原生肾脏和同种异体移植肾脏都有显著的有害影响。最近我们和其他研究小组在肾、肝、肺、肠和脑中的数据表明，淋巴细胞，特别是CD4 T细胞在IRI中起直接作用。T细胞通常对异体抗原或修饰的自身抗原有反应。迄今为止，引发IRI炎症和组织损伤的抗原尚不清楚。主要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，跨学科协同开展高风险，高影响的肾脏IRI新研究。假设：自身抗原在IRI中驱动T细胞的炎症和免疫反应中起重要作用。具体目标我们将使用已建立的小鼠肾夹诱导的温性IRI模型。我们将从IRI和假动物中获得肾周淋巴结，并使用hla II类免疫沉淀，分离IRI呈现的肽。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，我们将鉴定肾脏IRI期间参与T细胞免疫反应的蛋白质的特异性表位。目标2。我们将在Aim 1中鉴定的合成肽T细胞表位注射给幼稚小鼠，并在选择的时间，评估IRI的功能，组织学，分子和免疫学参数。我们将分离的合成肽加载到体外树突状细胞（dc）上，然后将这些负载的dc注射到小鼠体内。在注射多肽或负载DC后，小鼠将被提交IRI以确定保护水平和组织反应。预期结果：发现具有减少IRI组织损伤治疗潜力的新型多肽。