Skip to main content

Assessing NLRP3 Inflammasome Activation by Nanoparticles

Part of the Methods in Molecular Biology book series (MIMB,volume 1682)

Abstract

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.

Key words

  • Inflammasome
  • NLRP3
  • Nanoparticles
  • IL-1β
  • THP-1

This is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-4939-7352-1_12
  • Chapter length: 13 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-7352-1
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   139.99
Price excludes VAT (USA)
Hardcover Book
USD   179.99
Price excludes VAT (USA)
Fig. 1
Fig. 2
Fig. 3
Fig. 4

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Guo H, Callaway JB, Ting JPY (2015) Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med 21(7):677–687. doi:10.1038/nm.3893

    CrossRef  PubMed  PubMed Central  Google Scholar 

  2. Lund ME, To J, O'Brien BA, Donnelly S (2016) The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus. J Immunol Methods 430:64–70. doi:10.1016/j.jim.2016.01.012

    CAS  CrossRef  PubMed  Google Scholar 

  3. Kim M-G, Park JY, Shon Y, Kim G, Shim G, Oh Y-K (2014) Nanotechnology and vaccine development. Asian J Pharm Sci 9(5):227–235. http://dx.doi.org/10.1016/j.ajps.2014.06.002

    CrossRef  Google Scholar 

  4. Sharma D, Kanneganti TD (2016) The cell biology of inflammasomes: Mechanisms of inflammasome activation and regulation. J Cell Biol 213(6):617–629. doi:10.1083/jcb.201602089

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  5. Agostini L, Martinon F, Burns K, McDermott MF, Hawkins PN, Tschopp J (2004) NALP3 forms an IL-1beta-processing inflammasome with increased activity in Muckle-Wells autoinflammatory disorder. Immunity 20(3):319–325

    CAS  CrossRef  PubMed  Google Scholar 

  6. Mariathasan S, Weiss DS, Newton K, McBride J, O'Rourke K, Roose-Girma M, Lee WP, Weinrauch Y, Monack DM, Dixit VM (2006) Cryopyrin activates the inflammasome in response to toxins and ATP. Nature 440(7081):228–232. doi:10.1038/nature04515

    CAS  CrossRef  PubMed  Google Scholar 

  7. Meunier E, Coste A, Olagnier D, Authier H, Lefèvre L, Dardenne C, Bernad J, Béraud M, Flahaut E, Pipy B (2012) Double-walled carbon nanotubes trigger IL-1β release in human monocytes through Nlrp3 inflammasome activation. Nanomed Nanotechnol Biol Med 8(6):987–995

    Google Scholar 

  8. Luo Y.-H, Chang L.W, Lin P (2015) Metal-Based Nanoparticles and the Immune System: Activation, Inflammation, and Potential Applications. BioMed Res Int 2015:1–12

    Google Scholar 

  9. Okada M, Matsuzawa A, Yoshimura A, Ichijo H (2014) The lysosome rupture-activated TAK1-JNK pathway regulates NLRP3 inflammasome activation. J Biol Chem 289(47):32926–32936. doi:10.1074/jbc.M114.579961

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  10. Yaron JR, Gangaraju S, Rao MY, Kong X, Zhang L, Su F, Tian Y, Glenn HL, Meldrum DR (2015) K(+) regulates Ca(2+) to drive inflammasome signaling: dynamic visualization of ion flux in live cells. Cell Death Dis 6:e1954. doi:10.1038/cddis.2015.277

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  11. Ferrari D, Pizzirani C, Adinolfi E, Lemoli RM, Curti A, Idzko M, Panther E, Di Virgilio F (2006) The P2X7 receptor: a key player in IL-1 processing and release. J Immunol 176(7):3877–3883

    CAS  CrossRef  PubMed  Google Scholar 

  12. Tschopp J, Schroder K (2010) NLRP3 inflammasome activation: The convergence of multiple signalling pathways on ROS production? Nat Rev Immunol 10(3):210–215. doi:10.1038/nri2725

    CAS  CrossRef  PubMed  Google Scholar 

  13. Cullen SP, Kearney CJ, Clancy DM, Martin SJ (2015) Diverse activators of the NLRP3 inflammasome promote IL-1beta secretion by triggering necrosis. Cell Rep 11(10):1535–1548. doi:10.1016/j.celrep.2015.05.003

    CAS  CrossRef  PubMed  Google Scholar 

  14. Neumann S, Burkert K, Kemp R, Rades T, Rod Dunbar P, Hook S (2014) Activation of the NLRP3 inflammasome is not a feature of all particulate vaccine adjuvants. Immunol Cell Biol 92:535–542. 2014/04/02 edn. doi:10.1038/icb.2014.21

    CAS  CrossRef  PubMed  Google Scholar 

  15. Kusaka T, Nakayama M, Nakamura K, Ishimiya M, Furusawa E, Ogasawara K (2014) Effect of silica particle size on macrophage inflammatory responses. PLoS One 9(3):e92634. doi:10.1371/journal.pone.0092634

    CrossRef  PubMed  PubMed Central  Google Scholar 

  16. Reed SG, Orr MT, Fox CB (2013) Key roles of adjuvants in modern vaccines. Nat Med 19(12):1597–1608. doi:10.1038/nm.3409

    CAS  CrossRef  PubMed  Google Scholar 

  17. Lamkanfi M, Dixit VM (2014) Mechanisms and functions of inflammasomes. Cell 157(5):1013–1022. doi:10.1016/j.cell.2014.04.007

    CAS  CrossRef  PubMed  Google Scholar 

  18. van der Zande M, Vandebriel RJ, Groot MJ, Kramer E, Herrera Rivera ZE, Rasmussen K, Ossenkoppele JS, Tromp P, Gremmer ER, Peters RJ, Hendriksen PJ, Marvin HJ, Hoogenboom RL, Peijnenburg AA, Bouwmeester H (2014) Sub-chronic toxicity study in rats orally exposed to nanostructured silica. Part Fibre Toxicol 11(1):8. doi:10.1186/1743-8977-11-8

    CrossRef  PubMed  PubMed Central  Google Scholar 

  19. Neun BW, Dobrovolskaia MA (2011) Detection and quantitative evaluation of endotoxin contamination in nanoparticle formulations by LAL-based assays. Methods Mol Biol 697:121–130. doi:10.1007/978-1-60327-198-1_12

    CAS  CrossRef  PubMed  Google Scholar 

Download references

Acknowledgment

This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Pavan P. Adiseshaiah .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2018 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Sharma, B., McLeland, C.B., Potter, T.M., Stern, S.T., Adiseshaiah, P.P. (2018). Assessing NLRP3 Inflammasome Activation by Nanoparticles. In: McNeil, S. (eds) Characterization of Nanoparticles Intended for Drug Delivery. Methods in Molecular Biology, vol 1682. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7352-1_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7352-1_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7350-7

  • Online ISBN: 978-1-4939-7352-1

  • eBook Packages: Springer Protocols