Measurement of interleukin-17.
Upon antigenic stimulation, naive CD4+ T cells undergo proliferation and differentiate into cytokine-producing T helper (T(H)) effector cells. T(H)1 cells secrete effector cytokine IFN-gamma and regulate cell-mediated immunity, whereas T(H)2 cells produce IL-4, IL-5, and IL-13 cytokines, and mediate immunity against extracellular pathogens and allergic reactions. Recent studies have identified a novel T(H) subset, called T(H)17, TH(IL-17), or inflammatory T(H) (THi) cells, characterized by the production of a proinflammatory cytokine, IL-17, and regulating inflammatory responses. In this unit, we describe the protocols for the differentiation of mouse IL-17-expressing T cells in vitro, detection of IL-17-expressing T cells by intracellular cytokine staining, and measurement of IL-17 secretion in culture supernatants by ELISA. Generation of IL-17-expressing T cells in vitro under defined culture conditions allows investigation of their differentiation regulation. Detection of IL-17 in cell culture and tissue samples helps in monitoring inflammatory diseases and determining efficacy of therapeutic interventions.
Pappu
Bhanu P
BP
M.D. Anderson Cancer Center, Houston, Texas.
Dong
Chen
C
M.D. Anderson Cancer Center, Houston, Texas.
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D020381:Interleukin-17 / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D051379:Mice; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000378:metabolism*
D018836:Inflammation Mediators; D020381:Interleukin-17