Kuehn Hye Sun, Radinger Madeleine, Gilfillan Alasdair M
Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Curr Protoc Immunol. 2010 Nov;Chapter 7:Unit7.38. doi: 10.1002/0471142735.im0738s91.
Mediators released from activated mast cells are responsible for the allergic inflammatory reactions associated with disease states such as anaphylaxis and atopy. These mediators are released as a consequence of immediate degranulation and phospholipid metabolism upon mast cell activation, followed by delayed cytokine gene expression. Thus, techniques that monitor indices of these events in mast cell culture systems, in association with biochemical analysis of parameters of cell signaling, are critical to our understanding of the molecular mechanisms regulating mast cell-mediated disease. Furthermore, these systems can be adapted for high-throughput screens to identify potential inhibitors of mast cell activation that may provide potential leads for novel therapies for these diseases. In this unit, we describe approaches that can be readily used or adapted to a variety of rodent and human mast cell culture systems for the determination of degranulation, phospholipid-derived inflammatory mediator production, and cytokine generation.
活化肥大细胞释放的介质负责与诸如过敏反应和特应性等疾病状态相关的过敏性炎症反应。这些介质是肥大细胞活化后立即脱颗粒和磷脂代谢的结果，随后是细胞因子基因的延迟表达。因此，在肥大细胞培养系统中监测这些事件指标的技术，结合细胞信号参数的生化分析，对于我们理解调节肥大细胞介导疾病的分子机制至关重要。此外，这些系统可用于高通量筛选，以鉴定肥大细胞活化的潜在抑制剂，这些抑制剂可能为这些疾病的新疗法提供潜在线索。在本单元中，我们描述了可轻松用于或适用于各种啮齿动物和人类肥大细胞培养系统的方法，用于测定脱颗粒、磷脂衍生的炎症介质产生和细胞因子生成。
