| Flow Cytometry Measurement of Glucocerebrosidase Activity in Human Monocytes
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Author:
Date:
2020-04-05
[Abstract] Glucocerebrosidase (GCase) is an important enzyme for the metabolism of glycolipids. GCase enzyme deficiency is implicated in human disease and the efficient measurement of GCase activity is important for evaluating the efficacy of therapeutics targeting this enzyme. Existing approaches to measure GCase activity include whole blood mass spectrometry-based assays, where an internal standard is used to measure the accumulation of ceramide following metabolism of the synthetic substrate C12-glucocerebroside, and the utilisation of fluorescent probes that bind active GCase and/or release ...
[摘要]
[摘要 ] 葡萄糖脑苷脂酶（GCase ）是糖脂代谢的重要酶。GCase 酶缺乏症与人类疾病有关，GCase 活性的有效测量对于评估靶向该酶的治疗剂的功效至关重要。现有的测量GCase 活性的方法包括基于全血质谱的分析，其中使用内标测量合成底物C12-葡萄糖脑苷代谢后神经酰胺的积累，以及利用结合活性GCase 和// 的荧光探针的利用。或通过GCase 裂解后释放荧光代谢物 。在这里，我们描述了基于荧光激活细胞分选术的测定方法的应用，以有效地定量人类外周血单核细胞单核细胞群中的GCase 酶活性。细胞渗透性的GCase 衬底5-（Pentafluorobenzoylamino ）荧光素二-β-D- 吡喃葡萄糖苷（PFB- FDGlu ）提供了一个用于测量的GCase 活性，由此酶裂解产生的绿色荧光PFB-F染料，在FL-检测流式细胞仪的1个通道。使用溶酶体GCase 活性抑制剂，conduritol B-环氧，以确保特异性。该协议为测量活的单个细胞中的GCase 活性提供了一种有利的方法。
[背景 ] 葡萄糖脑苷脂酶（GCase ），由 GBA1基因是一种溶酶体水解酶，可将葡糖神经酰胺转化为葡萄糖和神经酰胺。GBA1 ...
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| T Cell Transfer Model of Colitis
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Author:
Date:
2016-07-05
[Abstract] Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC) is mainly caused by disordered immune regulation and dysregulated intestinal microbiota. Here we present the T cell transfer model which has extensively used in many studies to identify the regulatory T cell function in gut inflammation. Naïve T cells only or together with Treg cells isolated from different donors were transferred into immunodeficient Rag1-/- mice and the disease progression was assessed by the loss of body weight and the scoring analysis. This model provides a reliable ...
[摘要] 炎症性肠病（IBD），包括克罗恩病（CD）和溃疡性结肠炎（UC），主要由无序的免疫调节和肠道微生物群失调引起。 在这里我们提出的T细胞转移模型已广泛用于许多研究，以确定调节性T细胞功能的肠道炎症。 仅将来自不同供体的原始T细胞或与分离自不同供体的Treg细胞一起转移至免疫缺陷型Rag1 -/- 小鼠中，并通过体重损失和评分分析评估疾病进展。 这种模型为肠炎症的研究提供了可靠的工作系统。
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