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.
CITATION STYLE
Diamond, C., Bagnall, J., Spiller, D. G., White, M. R., Mortellaro, A., Paszek, P., & Brough, D. (2016). Investigating IL-1β secretion using real-time single-cell imaging. In Methods in Molecular Biology (Vol. 1417, pp. 75–88). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3566-6_4
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