Investigating IL-1ß Secretion Using Real-Time Single-Cell Imaging.
Investigating IL-1ß Secretion Using Real-Time Single-Cell Imaging.
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使用实时单细胞成像研究 IL-1 分泌。
DOI：
10.1007/978-1-4939-3566-6_4
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Diamond C
中科院分区：
文献类型：
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作者：
Diamond C
促炎细胞因子白细胞介素（IL）-1β是炎症反应的重要介质。为了在炎症级联反应中发挥作用，IL-1β必须从细胞中分泌，但它缺乏常规分泌所需的信号肽，并且释放的确切机制尚未确定。传统的生物化学方法将IL-1β分泌过程的研究局限于群体动力学，但在单细胞水平上观察到细胞间的异质性。在这里，通过使用新开发的编码荧光标记形式的IL-1β的载体实现了更高的灵敏度。将其与使用本文所述方法的实时单细胞共聚焦显微镜相结合，我们开发了一种有效的方案来研究IL-1β分泌的机制并测试IL-1β分泌需要膜透化的假设。
The pro-inflammatory cytokine interleukin (IL)-1β is an important mediator of the inflammatory response. In order to perform its role in the inflammatory cascade, IL-1β must be secreted from the cell, yet it lacks a signal peptide that is required for conventional secretion, and the exact mechanism of release remains undefined. Conventional biochemical methods have limited the investigation into the processes involved in IL-1β secretion to population dynamics, yet heterogeneity between cells has been observed at a single-cell level. Here, greater sensitivity is achieved with the use of a newly developed vector that codes for a fluorescently labelled version of IL-1β. Combining this with real-time single-cell confocal microscopy using the methods described here, we have developed an effective protocol for investigating the mechanisms of IL-1β secretion and the testing of the hypothesis that IL-1β secretion requires membrane permeabilisation.