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Immunology and Infection

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JoVE Journal Immunology and Infection
Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
 

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells

Article DOI: 10.3791/51284-v • 09:04 min • May 22nd, 2014
May 22nd, 2014 •
1, 2, 3
1Department of Pathology, New York University School of Medicine, 2Division of Infectious Diseases, Department of Medicine, Mount Sinai Medical Center, 3Division of Hematology and Oncology, Hess Center for Science and Medicine, Mount Sinai Medical Center

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Resumo
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Dendritic cells (DCs) secrete IL-1β in response to TLR8 recognition of synthetic purine, R848, followed by NLRP3 inflammasome activation with nigericin, therefore, IL-1β can be used to measure NLRP3 inflammasome activity. Intracellular cytokine staining, immunoblotting, and ELISA are used to accurately measure NLRP3 inflammasome priming and activation via IL-1β expression.

Tags

Activation Measurement NLRP3 Inflammasome Activity IL-1β Human Monocyte-derived Dendritic Cells Inflammatory Processes Interleukin-1 Family Cytokines Immune Cells Local Inflammation Systemic Inflammation Tissue Remodeling Virologic Control Innate Immune Response Invading Pathogens Persistent Infection Signaling Platform Interleukin 1 Converting Enzyme (ICE Or Caspase-1) Priming Signal Transcription Protein Expression Second Signal NLRP3 Inflammasome Formation Dendritic Cells Synthetic Purine R848 TLR8 Sensing Human Monocyte Derived Dendritic Cells (moDCs) Bacterial Toxin Potassium Ionophore Nigericin
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