Journal of Clinical and Cellular Immunology
Open Access
ISSN: 2155-9899
ISSN: 2155-9899
M R Dillingh
Posters: J Clin Cell Immunol
Inflammasomes are multiprotein complexes inducing pro-inflammatory cytokine production in response to infection and tissue injury. The end product of inflammasome activity is caspase-1-induced release of effector molecules such as IL-1 β and IL-18 from immune cells. Despite the common outcome, the variety of triggers that initiate inflammasome activity is large, and moreover eight different inflammasome subclasses have been described. The inflammasome is a potential therapeutic target for chronic inflammatory diseases such as chronic kidney disease and atherosclerosis. To test the effects of future inflammasome- targeted therapies, an ?inflammasome bioassay? is highly warranted. The inflammasome?s capacity to sense widely divergent ligands, and the diversity in inflammasome subclasses, indicates that a single bioassay may not be sufficient to assess potential inflammasome-modulating effects of new compounds. Therefore, we developed a set of bioassays inducing inflammasome activity, so-called ?inflammasome challenges?. Variables that were explored included matrix/cells (whole blood, PBMCs, THP1), triggers, incubation time (3-24 hours), dose-response relationship, single trigger versus immune cell priming followed by a second trigger, and inflammasome end products (IL-1β, IL-18). We successfully developed a set of inflammasome challenges, comprising inflammasome activation by LPS alone and by different triggers after LPS priming (extracellular ATP, aluminium hydroxide, cholesterol crystals, oligomeric β-amyloid). The short-term LPS inflammasome challenge was demonstrated to be sensitive to modulation of inflammasome activity, as assessed with a direct caspase-1 inhibitor. These inflammasome challenges can be applied in translational drug development as readout measures for inflammasome inhibition, and moreover will provide mechanistic insight in regulation of inflammasome activity.
Marlous Dillingh has a background in biomedical sciences. She is currently in training as a clinical pharmacologist at the Centre for Human Drug Research, Leiden, The Netherlands. Her main expertise is the integration of inflammation/immunology and pharmacology, applied in early phase clinical drug development, which is the focus of her PhD education.