Abstract:
The small molecule 8-methoxy-2-oxo-1,2,4,5-tetrahydrocyclopenta[de]quinoline-3-carboxylic acid (2b) behaves as a reactive non-fluorescent Michael acceptor, which after reaction with thiols becomes fluorescent, and an efficient Eu3+ antenna, after self-assembling with this cation in water. This behavior makes 2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8+ T, and B cells) using flow cytometry. GSH can be monitored by the fluorescence of the product of addition to 2b (445 nm) or by the luminescence of Eu3+ (592 nm). 2b was able to capture baseline differences in GSH intracellular levels among murine and human CD4+ T, CD8+ T, and B cells. We also successfully used 2b to monitor intracellular changes in GSH associated with the metabolic variations governing the induction of CD4+ naïve T cells into regulatory T cells (TREG).
摘要:
小分子8-甲氧基-2-氧代-1,2,4,5-四氢环戊[de]喹啉-3-羧酸(2b)表现为反应性非荧光迈克尔受体,与硫醇反应后变成荧光,和一个高效的Eu3+天线,在水中与这种阳离子自组装后。这种行为使2b成为高度选择性的GSH生物传感器,这已经证明了在免疫系统的鼠和人类细胞(CD4+T,CD8+T,和B细胞)使用流式细胞术。GSH可以通过添加到2b的产物的荧光(445nm)或通过Eu3+的发光(592nm)来监测。2b能够捕获小鼠和人CD4+T细胞内GSH水平的基线差异,CD8+T,B细胞。我们还成功地使用2b来监测与控制CD4+初始T细胞诱导成调节性T细胞(TREG)的代谢变化相关的GSH的细胞内变化。