Chapter title |
Human Primary Keratinocytes as a Tool for the Analysis of Caspase-1-Dependent Unconventional Protein Secretion
|
---|---|
Chapter number | 9 |
Book title |
Unconventional Protein Secretion
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3804-9_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3802-5, 978-1-4939-3804-9
|
Authors |
Gerhard E. Strittmatter, Martha Garstkiewicz, Jennifer Sand, Serena Grossi, Hans-Dietmar Beer |
Editors |
Andrea Pompa, Francesca De Marchis |
Abstract |
Inflammasomes comprise a group of protein complexes, which activate the protease caspase-1 upon sensing a variety of stress factors. Active caspase-1 in turn cleaves and thereby activates the pro-inflammatory cytokines prointerleukin (IL)-1β and -18, and induces unconventional protein secretion (UPS) of mature IL-1β, IL-18, as well as of many other proteins involved in and required for induction of inflammation. Human primary keratinocytes (HPKs) represent epithelial cells able to activate caspase-1 in an inflammasome-dependent manner upon irradiation with a physiological dose of ultraviolet B (UVB) light. Here, we describe the isolation of keratinocytes from human skin, their cultivation, and induction of caspase-1-dependent UPS upon UVB irradiation as well as its siRNA- and chemical-mediated inhibition. In contrast to inflammasome activation of professional immune cells, UVB-irradiated HPKs represent a robust and physiological cell culture system for the analysis of UPS induced by active caspase-1. |
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