Skip to main content

The Use of a Semi-Automated System to Measure Mouse Natural Killer T (NKT) Cell Activation by Lipid-Loaded Dendritic Cells

  • Protocol
  • First Online:
Food Allergens

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1592))

Abstract

Cell-based assays are widely used in all aspects of research ranging from understanding basic biological function to identifying compounds for disease intervention. Immortalized cell lines have been ideal components of these assays due to the low cost of growth, easy maintenance, and the ability to obtain homogenous cell populations. Like other molecular assays, cell-based systems have been automated to reduce experimental error. Complex lipids are now recognized as important components of the allergic response, the study of the interaction between NKTs and lipid-activated DCs opens a new perspective into the intrinsic allergenicity of proteins. Here, we describe a semi-automated method to measure IL-2 release upon activation of mouse NKT cells (DN32.D3) by various lipids in a 384-well plate using the Biomek® 3000 laboratory automated workstation (Beckman Coulter).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Amprey JL, Im JS, Turco SJ, Murray HW, Illarionov PA, Besra GS, Porcelli SA, Späth GF (2004) A subset of liver NK T cells is activated during Leishmania donovani infection by CD1d-bound lipophosphoglycan. J Exp Med 200(7):895–904

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Rhost S, Löfbom L, Rynmark BM, Pei B, Månsson JE, Teneberg S, Blomqvist M, Cardell SL (2012) Identification of novel glycolipid ligands activating a sulfatide-reactive, CD1d-restricted, type II natural killer T lymphocyte. Eur J Immunol 42(11):2851–2860

    Article  CAS  PubMed  Google Scholar 

  3. O'Konek JJ, Illarionov P, Khursigara DS, Ambrosino E, Izhak L, Castillo BF 2nd, Raju R, Khalili M, Kim HY, Howell AR, Besra GS, Porcelli SA, Berzofsky JA, Terabe M (2011) Mouse and human iNKT cell agonist β-mannosylceramide reveals a distinct mechanism of tumor immunity. J Clin Invest 121(2):683–694

    Article  PubMed  PubMed Central  Google Scholar 

  4. Mirotti L, Florsheim E, Rundqvist L, Larsson G, Spinozzi F, Leite-de-Moraes M, Russo M, Alcocer M (2013) Lipids are required for the development of Brazil nut allergy: the role of mouse and human iNKT cells. Allergy 68(1):74–83

    Article  CAS  PubMed  Google Scholar 

  5. Bashir ME, Lui JH, Palnivelu R, Naclerio RM, Preuss D (2013) Pollen lipidomics: lipid profiling exposes a notable diversity in 22 allergenic pollen and potential biomarkers of the allergic immune response. PLoS One 8(2):e57566

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Stangegaard M, Meijer PJ, Børsting C, Hansen AJ, Morling N (2012) Biomek 3000: the workhorse in an automated accredited forensic genetic laboratory. J Lab Autom 17(5):378–386

    Article  PubMed  Google Scholar 

  7. Jacques P, Venken K, Van Beneden K, Hammad H, Seeuws S, Drennan MB, Deforce D, Verbruggen G, Apostolaki M, Kollias G, Lambrecht BN, De Vos M, Elewaut D (2010) Invariant natural killer T cells are natural regulators of murine spondylarthritis. Arthritis Rheum 62(4):988–999

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ashfaq Ghumra .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Ghumra, A., Alcocer, M. (2017). The Use of a Semi-Automated System to Measure Mouse Natural Killer T (NKT) Cell Activation by Lipid-Loaded Dendritic Cells. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6925-8_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6923-4

  • Online ISBN: 978-1-4939-6925-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics