Analysis of IL-17 production by flow cytometry and ELISPOT assays

Methods Mol Biol. 2014:1172:243-56. doi: 10.1007/978-1-4939-0928-5_22.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / chemistry
  • Brefeldin A / pharmacology
  • Enzyme-Linked Immunospot Assay / methods*
  • Female
  • Flow Cytometry / methods*
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • Humans
  • Inflammation / immunology
  • Inflammation / pathology
  • Interleukin-17 / analysis*
  • Interleukin-17 / biosynthesis
  • Interleukin-17 / immunology
  • Interleukin-6 / pharmacology
  • Ionomycin / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Phycoerythrin
  • Quinolinium Compounds
  • Tetradecanoylphorbol Acetate / pharmacology
  • Th17 Cells / cytology
  • Th17 Cells / drug effects
  • Th17 Cells / immunology*
  • Transforming Growth Factor beta / pharmacology

Substances

  • Alexa fluor 546
  • Antibodies
  • Fluorescent Dyes
  • IL17A protein, human
  • Interleukin-17
  • Interleukin-6
  • Quinolinium Compounds
  • Transforming Growth Factor beta
  • Phycoerythrin
  • Brefeldin A
  • Ionomycin
  • Fluorescein-5-isothiocyanate
  • Tetradecanoylphorbol Acetate