Pyroptosis Detection via Multiplexing Cell Death and Bioluminescent Caspase-1 Activity Assessments
Poster Jul 14, 2017
Martha O’Brien1, Mike Scurria2, Tim Ugo2, Laurent Bernad2, James Cali1, and Dan Lazar1
Pyroptosis is a unique immunogenic form of cell death dependent on inflammasome activation, provoked when pattern recognition receptors (PRRs) sense microbial infection or cellular stress. Caspase-1, an essential component of inflammasomes, mediates this innate immune response by processing cytokines and eliciting cell death. In addition to caspase-1, other inflammatory caspases (caspase-11 in mice and caspases-4, 5 in humans) can cleave gasdermin D and elicit pyroptosis in response to some intracellular microbial infections. However, caspase-1 is also activated under these circumstances, therefore caspase-1 activation coincident with cell death is a reliable indicator of pyroptosis. Understanding when PRR sensing leads to cell death and when it does not is essential to understanding the complexities of the innate immune response. We have developed a convenient method to monitor pyroptosis by multiplexing in the same assay well our homogeneous, bioluminescent caspase-1 activity assay with a membrane-impermeant, stable DNA dye for the detection of cell death. In contrast to endpoint assays for cell death such as lactate dehydrogenase (LDH) assays, the DNA dye can be used in real-time. Cell death can be monitored over time and when first detected, the caspase-1 assay can be performed confirming pyroptosis. Alternatively, culture medium can be removed for testing for caspase-1 release, leaving the cells available for measuring cell death by a variety of means. We show examples of pyroptosis in human THP-1 monocytes and mouse J774A.1 macrophages.
Extracorporeal shockwave therapy accelerates motor axon regeneration despite a phenotypically mismatched environmentPoster
A femoral nerve defect model was adapted for the evaluation of proregenerative effects of extracorporeal shockwave therapy (ESWT). Functional evaluation, histology and qRT-PCR data show differences between sensory and motor-derived nerve transplants and a pro-regenerative effect of ESWT. These data provide evidence for the clinical application of ESWT after autologous nerve transplantation as a novel non-invasive method.READ MORE
Histone Deacetylase 6 (HDAC6) As A Therapeutic Target in Chronic Lymphocytic LeukemiaPoster
This study investigates the role of histone deacetylase 6 (HDAC6) in chronic lymphocytic leukemia and establishes it as a novel therapeutic target for the treatment of this disease.READ MORE