PRECLINICAL MODEL OF CHRONIC RENAL ALLOGRAFT REJECTION

移植免疫抑制治疗的进展 几乎没有降低同种异体移植物的百分比， 最终发展为慢性排斥反应（CR）。 拟议的研究 本申请共同作为临床前桥梁， 与器官移植有关。 实验设计 在这个假设的框架内， 与超氧化物一氧化氮相关的氧化应激， 反应产物，过氧亚硝酸根，酸性成纤维细胞生长因子 （FGF-1）介导慢性肾脏病期间的转化潜力 通过细胞外途径的同种异体移植排斥。 下面的 这些研究的主题包括量化生产和 活性氮/氧物质的靶向分子反应 抗氧化防御机制（锰 超氧化物歧化酶和谷胱甘肽）在肾移植中的作用， 损害肾结构/功能。 设立：（a）a 氧化应激、FGF-1和 CR：（B）用于监测该过程的诊断标准;以及（c） 治疗性干预策略， 抗FGF-1和抗氧化剂的中和抗体将被 评价它们调节发育动力学的能力， 建立的同种异体大鼠肾脏模型中的CR病变 移植 抗体、核酸、 探针/扩增器，和建立的技术允许 通过分析解析CR期间的响应分子级联 mRNA（RT-PCR，杂交）和蛋白质（Western，ELISA， 免疫组织化学，酶活性）在肾中的表达。 特别强调将集中在FGF-1的调节能力， 增殖细胞核抗原（PCNA），过氧亚硝基诱导的凋亡（TUNEL）， 靶蛋白中的酪氨酸硝化。 泌尿系统相关性 FGF-1、亚硝酸盐/硝酸盐、白蛋白和硝基酪氨酸的出现 将形成具有原位发展的病理损伤的蛋白质 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。 虽然确切的参与 FGF-1，活性氮/氧物质，抗氧化剂，和 硝基酪氨酸蛋白是未知的，本申请中的努力 将阐明反映病理生理学的详细特征， 与CR相关的过程不仅在大鼠模型中， 接受肾移植的灵长类动物和人类患者 免疫抑制疗法。

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.