Abstract
Interleukin (IL)-17 represents a family of cytokines with six members, namely IL-17A, B, C, D, E, and F. IL-17A and IL-17F are best studied proinflammatory cytokines. CD4+ T helper cells producing IL-17A have been identified as a distinct T helper subset, Th17 cells. IL-17 and Th17 cells are important mediators in tissue inflammation in immune-mediated inflammatory diseases. IL-17 is also produced by other immune cells and plays an important role in host defense against microbial infection. Cell-based assays are sensitive and quantitative, and enable identification of cellular sources of IL-17 production. This chapter describes usage of flow cytometry and ELISPOT assays to quantify IL-17A-producing cells in disease and in vitro experiments to study T cell function.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rouvier E, Luciani MF, Mattei MG et al (1993) CTLA-8, cloned from an activated T cell, bearing AU-rich messenger RNA instability sequences, and homologous to a herpesvirus saimiri gene. J Immunol 150:5445–5456
Kolls JK, Linden A (2004) Interleukin-17 family members and inflammation. Immunity 21:467–476
Miossec P, Kolls JK (2012) Targeting IL-17 and TH17 cells in chronic inflammation. Nat Rev Drug Discov 11:763–776
Wright JF, Guo Y, Quazi A et al (2007) Identification of an interleukin 17F/17A heterodimer in activated human CD4+ T cells. J Biol Chem 282:13447–13455
Aggarwal S, Ghilardi N, Xie MH et al (2003) Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17. J Biol Chem 278:1910–1914
Harrington LE, Hatton RD, Mangan PR et al (2005) Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol 6:1123–1132
Park H, Li Z, Yang XO et al (2005) A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol 6:1133–1141
Veldhoen M, Hocking RJ, Atkins CJ et al (2006) TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells. Immunity 24:179–189
Acosta-Rodriguez EV, Napolitani G, Lanzavecchia A et al (2007) Interleukins 1beta and 6 but not transforming growth factor-beta are essential for the differentiation of interleukin 17-producing human T helper cells. Nat Immunol 8:942–949
Murphy CA, Langrish CL, Chen Y et al (2003) Divergent pro- and antiinflammatory roles for IL-23 and IL-12 in joint autoimmune inflammation. J Exp Med 198:1951–1957
Cua DJ, Sherlock J, Chen Y et al (2003) Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain. Nature 421:744–748
Haines CJ, Chen Y, Blumenschein WM et al (2013) Autoimmune memory T helper 17 cell function and expansion are dependent on interleukin-23. Cell Rep 3:1378–1388
Martin B, Hirota K, Cua DJ et al (2009) Interleukin-17-producing gammadelta T cells selectively expand in response to pathogen products and environmental signals. Immunity 31:321–330
Cho JS, Pietras EM, Garcia NC et al (2010) IL-17 is essential for host defense against cutaneous Staphylococcus aureus infection in mice. J Clin Invest 120:1762–1773
Puel A, Cypowyj S, Bustamante J et al (2011) Chronic mucocutaneous candidiasis in humans with inborn errors of interleukin-17 immunity. Science 332:65–68
Yang XO, Chang SH, Park H et al (2008) Regulation of inflammatory responses by IL-17F. J Exp Med 205:1063–1075
Karlsson AC, Martin JN, Younger SR et al (2003) Comparison of the ELISPOT and cytokine flow cytometry assays for the enumeration of antigen-specific T cells. J Immunol Methods 283:141–153
Anthony DD, Lehmann PV (2003) T-cell epitope mapping using the ELISPOT approach. Methods 29:260–269
Zhao L, Jiang Z, Jiang Y et al (2013) IL-22(+) CD4(+) T cells in patients with rheumatoid arthritis. Int J Rheum Dis 16:518–526
Zhao L, Jiang Z, Jiang Y et al (2013) IL-22+CD4+ T-cells in patients with active systemic lupus erythematosus. Exp Biol Med (Maywood) 238:193–199
Morita Y, Ismail DM, Elkon KB et al (2011) Dichotomous response to transforming growth factor beta after T cell receptor activation by naive CD4+ T cells from DBA/1 mice: enhanced retinoic acid receptor-related orphan nuclear receptor gammat expression yet reduced FoxP3 expression. Arthritis Rheum 63:118–126
Yomogida K, Chou YK, Chu CQ (2013) Superantigens induce IL-17 production from polarized Th1 clones. Cytokine 63:6–9
Zhu J, Paul WE (2010) CD4+ T cell plasticity-Th2 cells join the crowd. Immunity 32:11–13
Chou YK, Henderikx P, Vainiene M et al (1991) Specificity of human T cell clones reactive to immunodominant epitopes of myelin basic protein. J Neurosci Res 28:280–290
Acknowledgement
This work was supported by grants from the National Institutes of Health AR-055254 (CQC) and the National Natural Science Foundation of China (No. 30972610) (YJ and ZJ).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Zhao, L., Chou, Y., Jiang, Y., Jiang, Z., Chu, CQ. (2014). Analysis of IL-17 Production by Flow Cytometry and ELISPOT Assays. In: Vancurova, I. (eds) Cytokine Bioassays. Methods in Molecular Biology, vol 1172. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0928-5_22
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0928-5_22
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0927-8
Online ISBN: 978-1-4939-0928-5
eBook Packages: Springer Protocols