CELL MEDIATED RENAL INJURY IN LUPUS

狼疮细胞介导的肾损伤

• 批准号: 3234497
• 负责人: Vicki Rubin Kelley
• 金额: $ 13.96万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-02-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3234497
• 关键词: arachidonate; autoimmune disorder; cellular immunity; connective tissue disorder chemotherapy; cytokine; eicosanoid metabolism; fluorescence microscopy; gene expression; genetic strain; histocompatibility antigens; immunocytochemistry; immunogenetics; immunopharmacology; in situ hybridization; interleukin 1; ionophores; laboratory mouse; lymphokines; macrophage; marine animal oil; mesangium; nephritis; northern blottings; nucleic acid probes; nutrition related tag; omega 3 fatty acid; radioimmunoassay; systemic lupus erythematosus; tissue /cell culture; tumor necrosis factor alpha

The broad objective of this proposal is to examine the relationship between autoimmune mediated renal injury and the activation of cytokine genes in invading macrophages (macs) in the kidney and intrinsic renal cells. We plan to dissect the kinetic and functional relationship between these events. Cellular and molecular approaches will be used to examine two spontaneous murine models of autoimmune lupus nephritis, the MRL-lpr and NZBxNZW F1 hybrid mice . Renal injury will be determined morphologically, immunohistochemically and functionally. Gene expression for cytokines will be analyzed by Northern blots and quantitated by laser densitometry and cytokine production detected by bioassays and radioimmunassays. Analysis will extend from the whole kidney to isolated compartments including cortex, medulla, glomeruli and cultured and transformed glomerular macs and mesangial cells. Using in-situ hybridization techniques cytokine mRNA transcripts will be detected in intrinsic renal cells. The specific aims define the temporal sequence of an increase in macs, interleukin 1 (IL-I) and tumor necrosis factor-(TNF) in the pathogenesis of lupus nephritis. Experiments will characterize and define the function of glomerular macs in lupus nephritis. These macs which accumulate in the mesangium are morphologically distinctive since they are laden with electron dense deposits. In addition, in culture glomerular macs are unique since they are activated, readily proliferate, and express IL-3 in addition to the cytokines characteristic of macs, IL-lB, IL-l and TNF. Using cultured and Sv40 transformed glomerular macs, studies will examine cytokine gene expression, production and regulation and determine if these macs are antigen presenters. Adoptive transfer and depletion experiments will examine the role of macs in renal injury. Finally, we will determine if IL-l attracts these macs and whether activation occurs in extra-renal sites or within the kidney. The role of IL-l and TNF in renal disease will be examined by diverse strategies that suppress and accelerate lupus nephritis, and then by measuring and correlating the level of IL-l and TNF gene expression. Conversely, IL-l and TNF and their antibodies will be administered and the progression of renal injury evaluated. Taken together these studies should clarify the importance of cytokines and macs in the progression of lupus nephritis.

该提议的广泛目标是检查关系 在自身免疫性介导的肾脏损伤和激活之间 肾脏入侵巨噬细胞（MAC）中的细胞因子基因和 固有的肾细胞。 我们计划剖析动力学和 这些事件之间的功能关系。 细胞和 分子方法将用于检查两种自发鼠 自身免疫性狼疮肾炎，MRL-LPR和NZBXNZW F1的模型 混合小鼠。 肾脏损伤将在形态上确定， 免疫组织化学和功能。 基因表达 细胞因子将通过北印迹分析，并通过 生物测定检测到的激光密度法和细胞因子产生 和放射膜。 分析将从整个肾脏延伸 到包括皮质，髓质，肾小球和 培养和转化的肾小球MAC和细胞。 使用原位杂交技术细胞因子mRNA转录本 将在固有的肾细胞中检测到。 具体目标 定义MAC的增加的时间顺序 1（IL-I）和肿瘤坏死因子（TNF） 狼疮肾炎。 实验将表征并定义 肾小球肾小球肾炎中肾小球的功能。 这些Mac 在肾小肌中积聚在形态上是独特的，因为 他们载有电子致密沉积物。 另外，在 培养肾小球Mac是独一无二的，因为它们被激活， 除了细胞因子之外，很容易增殖并表达IL-3 Mac，IL-LB，IL-L和TNF的特征。 使用培养和 SV40转化的肾小球MAC，研究将检查细胞因子 基因表达，生产和调节，并确定是否 Mac是抗原演示者。 收养和耗尽 实验将检查MAC在肾脏损伤中的作用。 最后，我们将确定IL-L是否吸引了这些Mac以及是否吸引 激活发生在肾外部位或肾脏内。 这 IL-L和TNF在肾脏疾病中的作用将通过多种 抑制和加速狼疮肾炎的策略，然后 通过测量和关联IL-L和TNF基因的水平 表达。 相反，IL-L和TNF及其抗体将是 管理和评估肾脏损伤的进展。 拍摄 这些研究共同阐明了细胞因子的重要性 和狼疮肾炎进展的MAC。