Skip to main content

Cytokine Multiplex Immunoassay: Methodology and (Clinical) Applications

  • Protocol
T Cell Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 514))

Abstract

Subsets of T cells can be distinguished on basis of their cytokine production and secretion profile.

With the critical role of cytokines in the regulation of immune and inflammatory responses, cytokines hold the promise to become the ideal biomarkers to monitor development and progression of immune-mediated diseases, study the development of new therapeutic approaches (both in vitro and in vivo) and as outcome parameters. Because of the numerous interactions in the cytokine network, the pleiotropic actions and redundancy, it will be necessary to monitor the complete spectrum of cytokines. As such, the multiplex immunoassay (MIA) is the ideal technique for that purpose. This paper reviews the critical methodological steps of this technique, including the procedures for antibody coupling to beads and matrix effects of biological fluids and buffer solutions. The power and robustness of the MIA technique is illustrated by an analysis of cytokine profiles in juvenile idiopathic arthritis.

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 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Park H, Li Z, Yang XO, et al. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol 2005;6:1133–41.

    Article  CAS  PubMed  Google Scholar 

  2. Harrington LE, Hatton RD, Mangan PR, et al. Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T-helper type 1 and 2 lineages. Nat Immunol 2005;6:1123–32.

    Article  CAS  PubMed  Google Scholar 

  3. de Jager W, Prakken BJ, Bijlsma JW, Kuis W, Rijkers GT. Improved multiplex immunoassay performance in human plasma and synovial fluid following removal of interfering heterophilic antibodies. J Immunol Methods 2005;300:124–35.

    Article  PubMed  Google Scholar 

  4. de Jager W, Hoppenreijs EP, Wulffraat NM, Wedderburn LR, Kuis W, Prakken BJ. Blood and synovial fluid cytokine signatures in patients with juvenile idiopathic arthritis: a cross-sectional study. Ann Rheum Dis 2007;66:589–98.

    Article  PubMed  Google Scholar 

  5. Jiang H, Chess L. Regulation of immune responses by T cells. N Engl J Med 2006;354:1166–76.

    Article  CAS  PubMed  Google Scholar 

  6. Hori S, Takahashi T, Sakaguchi S. Control of autoimmunity by naturally arising regulatory CD4+ T cells. Adv Immunol 2003;81:331–71.

    Article  CAS  PubMed  Google Scholar 

  7. Sakaguchi S. Regulatory T cells: Meden Agan Immunol Rev 2006;212:5–7.

    Google Scholar 

  8. Sakaguchi S, Sakaguchi N, Asano M, Itoh M, Toda M. Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol 1995;155:1151–64.

    CAS  PubMed  Google Scholar 

  9. Vignali DA. Multiplexed particle-based flow cytometric assays. J Immunol Methods 2000;243:243–55.

    Article  CAS  PubMed  Google Scholar 

  10. Motulsky HJ, Ransnas LA. Fitting curves to data using nonlinear regression: a practical and nonmathematical review. FASEB J 1987;1:365–74.

    CAS  PubMed  Google Scholar 

  11. Baud M. Data analysis, mathematical modeling. In: Masseyeff RF, Albert W, Staines NA, eds. Methods of Immunological Analysis Volume 1: Fundamentals. New York: VCH Publishers Inc, 1993:656–71

    Google Scholar 

  12. Staros JV, Wright RW, Swingle DM. Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem 1986;156:220–2.

    Article  CAS  PubMed  Google Scholar 

  13. Grabarek Z, Gergely J. Zero-length crosslinking procedure with the use of active esters. Anal Biochem 1990;185:131–5.

    Article  CAS  PubMed  Google Scholar 

  14. Timkovich R. Detection of the stable addition of carbodiimide to proteins. Anal Biochem 1977;79:135–43.

    Article  CAS  PubMed  Google Scholar 

  15. de Jager W, Rijkers GT. Solid-phase and bead-based cytokine immunoassay: a comparison. Methods 2006;38:294–303.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

de Jager, W., Prakken, B., Rijkers, G.T. (2009). Cytokine Multiplex Immunoassay: Methodology and (Clinical) Applications. In: De Libero, G. (eds) T Cell Protocols. Methods in Molecular Biology™, vol 514. Humana Press. https://doi.org/10.1007/978-1-60327-527-9_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-527-9_9

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-587-3

  • Online ISBN: 978-1-60327-527-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics