PRECLINICAL MODEL OF CHRONIC RENAL ALLOGRAFT REJECTION

慢性同种异体肾移植排斥的临床前模型

• 批准号: 2017362
• 负责人: JOHN A THOMPSON
• 金额: $ 22.14万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-22 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2017362
• 关键词: antibody; antioxidants; apoptosis; biological models; enzyme linked immunosorbent assay; fibroblast growth factor; glutathione; human subject; immunocytochemistry; kidney transplantation; laboratory rat; model design /development; nitric oxide; nucleic acid hybridization; oxidative stress; polymerase chain reaction; superoxide dismutase; superoxides; transplant rejection; western blottings

Advances in immunosuppressive therapy for transplantation have done little to decrease the percentage of allografts which eventually develop chronic rejection (CR). Studies proposed in this application collectively serve as a preclinical bridge with direct relevance to organ transplantation. The experiments are designed within the framework of the hypothesis that in response to oxidative stress associated with superoxide nitric oxide, and their reaction product, peroxynitrite, acidic fibroblast growth factor (FGF-1) mediates a transforming potential during chronic renal allograft rejection by an extracellular pathway. An underlying theme of these studies includes quantitating the production and targeted molecular reactions of reactive nitrogen/oxygen species that inactivate antioxidant defense mechanisms (manganese superoxide dismutase and glutathione) in the kidney allograft to compromise renal structure/function. To establish: (a) a cause-and-effect relationship between oxidant stress, FGF-1, and CR: (b) a diagnostic criteria for monitoring this process; and (c) therapeutic interventional strategies, the administration of neutralizing antibodies against FGF-1 and antioxidants will be evaluated for their ability to modulate the kinetics of developing CR lesions in an established allogeneic rat model of kidney transplantation. The availability of antibodies, nucleic acid probel/amplimers, and established techniques permits the dissection of responding molecular cascades during CR by analyses of mRNA (RT-PCR, hybridization) and protein (Western, ELISA, immunohistochemistry, enzymatic activity) expression in the kidney. Special emphasis will focus on the ability of FGF-1 to modulate growth (PCNA), peroxynitrite-induced apoptosis (TUNEL), and tyrosine nitration in target proteins. The correlation of urinary appearance of FGF-1, nitrite/nitrate, albumin, and nitrotyrosyl proteins with in situ development of pathologic lesions will form the basis of a non-invasive diagnostic assay for monitoring CR during renal transplant dysfunction. While the precise involvement of FGF-1, reactive nitrogen/oxygen species, antioxidants, and nitrotyrosyl proteins are not known, efforts within this application will elucidate detailed characteristics reflective of pathophysiologic processes associated with CR not only in the rat model but also in kidney transplanted primates and human patients undergoing immunosuppressive therapies.

免疫抑制治疗的移植疗法的进步已有 做几乎没有降低同种异体移植的百分比 最终发展慢性排斥（CR）。 研究提出了 该应用程序集体充当直接的临床前桥 与器官移植相关。 实验是设计的 在回应的假设的框架内 与超氧化氧化氮相关的氧化应激及其 反应产物，过氧亚硝酸盐，酸性成纤维细胞生长因子 （FGF-1）介导慢性肾脏期间的转化潜力 细胞外途径排斥反应。 基础 这些研究的主题包括量化生产和 活性氮/氧的靶向分子反应 灭活抗氧化剂防御机制（锰 肾脏同种异体移植中的超氧化物歧化酶和谷胱甘肽 妥协的肾脏结构/功能。 建立：（a）a 氧化应激，FGF-1和 CR：（b）用于监视此过程的诊断标准； （c） 治疗性介入策略，管理 对FGF-1和抗氧化剂的中和抗体将是 评估他们调节开发动力学的能力 肾脏的同种异体大鼠模型中的CR病变 移植。 抗体，核酸的可用性 Probel/Amplimers，并建立的技术允许 通过分析在CR期间对响应分子级联反应的解剖 mRNA（RT-PCR，杂交）和蛋白质（Western，Elisa， 免疫组织化学，酶活性）在肾脏中的表达。 特别重点将重点放在FGF-1调制的能力上 生长（PCNA），过氧亚硝酸盐诱导的凋亡（TUNEL）和 靶蛋白中的酪氨酸硝化。 尿的相关性 FGF-1，亚硝酸盐/硝酸盐，白蛋白和硝基三烷基的外观 具有病理病变原位发展的蛋白质将形成 用于监测CR的非侵入性诊断测定的基础 在肾移植功能障碍期间。 而精确的参与 FGF-1，反应性氮/氧，抗氧化剂和 硝基三酰基蛋白尚不清楚，在此应用中的努力 将阐明反映病理生理的详细特征 不仅在大鼠模型中，而且在 肾脏移植的灵长类动物和人类患者 免疫抑制疗法。