From the Divisions of Immunology and.
J Biol Chem. 2014 Apr 4;289(14):9730-40. doi: 10.1074/jbc.M113.538041. Epub 2014 Feb 25.
The cysteine protease inhibitor cystatin C is thought to be secreted by most cells and eliminated in the kidneys, so its concentration in plasma is diagnostic of kidney function. Low extracellular cystatin C is linked to pathologic protease activity in cancer, arthritis, atherosclerosis, aortic aneurism, and emphysema. Cystatin C forms non-inhibitory dimers and aggregates by a mechanism known as domain swapping, a property that reportedly protects against Alzheimer disease but can also cause amyloid angiopathy. Despite these clinical associations, little is known about the regulation of cystatin C production, dimerization, and secretion. We show that hematopoietic cells are major contributors to extracellular cystatin C levels in healthy mice. Among these cells, macrophages and dendritic cells (DC) are the predominant producers of cystatin C. Both cell types synthesize monomeric and dimeric cystatin C in vivo, but only secrete monomer. Dimerization occurs co-translationally in the endoplasmic reticulum and is regulated by the levels of reactive oxygen species (ROS) derived from mitochondria. Drugs or stimuli that reduce the intracellular concentration of ROS inhibit cystatin C dimerization. The extracellular concentration of inhibitory cystatin C is thus partly dependent on the abundance of macrophages and DC, and the ROS levels. These results have implications for the diagnostic use of serum cystatin C as a marker of kidney function during inflammatory processes that induce changes in DC or macrophage abundance. They also suggest an important role for macrophages, DC, and ROS in diseases associated with the protease inhibitory activity or amyloidogenic properties of cystatin C.
半胱氨酸蛋白酶抑制剂胱抑素 C 被认为由大多数细胞分泌,并在肾脏中被清除,因此其在血浆中的浓度可用于诊断肾脏功能。细胞外胱抑素 C 水平低与癌症、关节炎、动脉粥样硬化、主动脉瘤和肺气肿中的病理性蛋白酶活性有关。胱抑素 C 通过一种称为结构域交换的机制形成非抑制性二聚体和聚集体,据报道,这种特性可以预防阿尔茨海默病,但也可能导致淀粉样血管病。尽管有这些临床关联,但人们对胱抑素 C 产生、二聚化和分泌的调节知之甚少。我们表明,造血细胞是健康小鼠细胞外胱抑素 C 水平的主要贡献者。在这些细胞中,巨噬细胞和树突状细胞 (DC) 是胱抑素 C 的主要产生者。这两种细胞类型都在体内合成单体和二聚体胱抑素 C,但仅分泌单体。二聚化发生在内质网中,并受来自线粒体的活性氧 (ROS) 水平的调节。降低细胞内 ROS 水平的药物或刺激物会抑制胱抑素 C 的二聚化。因此,抑制性胱抑素 C 的细胞外浓度部分取决于巨噬细胞和 DC 的丰度以及 ROS 水平。这些结果对诊断使用血清胱抑素 C 作为炎症过程中肾功能的标志物具有重要意义,这些过程会引起 DC 或巨噬细胞丰度的变化。它们还表明,巨噬细胞、DC 和 ROS 在与胱抑素 C 的蛋白酶抑制活性或淀粉样特性相关的疾病中具有重要作用。