Assessing NLRP3 Inflammasome Activation by Nanoparticles

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1682; p. 135
• Main Authors: Sharma, Bhawna, McLeland, Christopher B, Potter, Timothy M, Stern, Stephan T, Adiseshaiah, Pavan P
• Format: Journal Article
• Language: English
• Published: United States 2018
• Subjects: Cell Culture Techniques - methods; Cell Line; Cell Survival; Enzyme-Linked Immunosorbent Assay - methods; Humans; Inflammasomes - immunology; Interleukin-1beta - immunology; Nanoparticles - adverse effects; NLR Family, Pyrin Domain-Containing 3 Protein - immunology; Nanoparticles; THP-1; NLRP3; Inflammasome; IL-1β
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-7352-1_12

NLRP3 inflammasome activation is one of the initial steps in an inflammatory cascade against pathogen/danger-associated molecular patterns (PAMPs/DAMPs), such as those arising from environmental toxins or nanoparticles, and is essential for innate immune response. NLRP3 inflammasome activation in cells can lead to the release of IL-1β cytokine via caspase-1, which is required for inflammatory-induced programmed cell death (pyroptosis). Nanoparticles are commonly used as vaccine adjuvants and drug delivery vehicles to improve the efficacy and reduce the toxicity of chemotherapeutic agents. Several studies indicate that different nanoparticles (e.g., liposomes, polymer-based nanoparticles) can induce NLRP3 inflammasome activation. Generation of a pro-inflammatory response is beneficial for vaccine delivery to provide adaptive immunity, a necessary step for successful vaccination. However, similar immune responses for intravenously injected, drug-containing nanoparticles can result in immunotoxicity (e.g., silica nanoparticles). Evaluation of NLRP3-mediated inflammasome activation by nanoparticles may predict pro-inflammatory responses in order to determine if these effects may be mitigated for drug delivery or optimized for vaccine development. In this protocol, we outline steps to monitor the release of IL-1β using PMA-primed THP-1 cells, a human monocytic leukemia cell line, as a model system. IL-1β release is used as a marker of NLRP3 inflammasome activation.