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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
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.
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.
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.
Jiang H, Chess L. Regulation of immune responses by T cells. N Engl J Med 2006;354:1166–76.
Hori S, Takahashi T, Sakaguchi S. Control of autoimmunity by naturally arising regulatory CD4+ T cells. Adv Immunol 2003;81:331–71.
Sakaguchi S. Regulatory T cells: Meden Agan Immunol Rev 2006;212:5–7.
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.
Vignali DA. Multiplexed particle-based flow cytometric assays. J Immunol Methods 2000;243:243–55.
Motulsky HJ, Ransnas LA. Fitting curves to data using nonlinear regression: a practical and nonmathematical review. FASEB J 1987;1:365–74.
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
Staros JV, Wright RW, Swingle DM. Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem 1986;156:220–2.
Grabarek Z, Gergely J. Zero-length crosslinking procedure with the use of active esters. Anal Biochem 1990;185:131–5.
Timkovich R. Detection of the stable addition of carbodiimide to proteins. Anal Biochem 1977;79:135–43.
de Jager W, Rijkers GT. Solid-phase and bead-based cytokine immunoassay: a comparison. Methods 2006;38:294–303.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights 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