Kisilevsky Robert
Department of Pathology and Molecular Medicine, Queen's University and The Syl and Molly Apps Research Center, Kingston General Hospital, Ontario, Canada.
Methods Mol Biol. 2005;299:237-41. doi: 10.1385/1-59259-874-9:237.
Amyloid-enhancing factor (AEF) is a biological "activity" that is defined in the context of inflammation-associated amyloidogenesis (AA). When administered intravenously to mice followed by an inflammatory stimulus, such primed mice deposit substantial AA amyloid in spleen within 36-48 h. Since experimental induction of AEF is dependent on amyloidogenic protocols, and rapid AA amyloid induction is dependent on AEF, a strategy for AEF isolation is required to break into this circular process. AEF activity may be prepared from a variety of human forms of amyloid that include tissue containing any of Abeta, AA, ATTR, and AL amyloids. The preparation of an AEF extract from such human tissue is described using 4 M glycerol, which then may be used to induce splenic AA amyloid fibrils in mice as a source for the propagation of additional AEF and/or for the study of amyloidogenesis. The glycerol and AA fibril preparations are stable frozen for many years.
淀粉样增强因子(AEF)是一种生物学“活性物质”,其定义与炎症相关的淀粉样蛋白生成(AA)有关。给小鼠静脉注射AEF后再给予炎症刺激,这种预处理的小鼠在36 - 48小时内会在脾脏中沉积大量的AA淀粉样蛋白。由于AEF的实验诱导依赖于淀粉样蛋白生成方案,而快速的AA淀粉样蛋白诱导依赖于AEF,因此需要一种AEF分离策略来打破这个循环过程。AEF活性物质可从多种人类淀粉样蛋白形式中制备,这些形式包括含有β淀粉样蛋白(Aβ)、AA淀粉样蛋白、转甲状腺素蛋白淀粉样变(ATTR)和轻链淀粉样变(AL)淀粉样蛋白的组织。本文描述了使用4M甘油从这种人类组织中制备AEF提取物的方法,然后该提取物可用于在小鼠中诱导脾脏AA淀粉样纤维,作为额外AEF繁殖的来源和/或用于淀粉样蛋白生成的研究。甘油和AA纤维制剂在冷冻状态下可稳定保存多年。