Human Primary Keratinocytes as a Tool for the Analysis of Caspase-1-Dependent Unconventional Protein Secretion.
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.
Department of Dermatology, University Hospital Zurich, University Zurich, Gloriastrasse 31, CH-8091, Zurich, Switzerland.
Gerhard E
GE
Strittmatter
Department of Dermatology, University Hospital Zurich, University Zurich, Gloriastrasse 31, CH-8091, Zurich, Switzerland.
Martha
M
Garstkiewicz
Department of Dermatology, University Hospital Zurich, University Zurich, Gloriastrasse 31, CH-8091, Zurich, Switzerland.
Jennifer
J
Sand
Department of Dermatology, University Hospital Zurich, University Zurich, Gloriastrasse 31, CH-8091, Zurich, Switzerland.
Serena
S
Grossi
Department of Dermatology, University Hospital Zurich, University Zurich, Gloriastrasse 31, CH-8091, Zurich, Switzerland. Hans-Dietmar.Beer@usz.ch.
Hans-Dietmar
HD
Beer
D020170:Caspase 1 / Q000378:metabolism*; D002469:Cell Separation; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006801:Humans; D058847:Inflammasomes / Q000378:metabolism; D007375:Interleukin-1 / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D055571:Secretory Pathway; D014162:Transfection
D058847:Inflammasomes; D007375:Interleukin-1; D011506:Proteins; D034741:RNA, Small Interfering; D020170:Caspase 1
Caspase-1; Inflammasomes; Interleukin-1; Keratinocytes; UVB irradiation; Unconventional protein secretion