title: Isolation and th17 differentiation of naïve CD4 T lymphocytes.
abstract: Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets including Th1, Th2, and regulatory T cells. Th17 cells have emerged as a central culprit in overzealous inflammatory immune responses associated with many autoimmune disorders. In this method we purify T lymphocytes from the spleen and lymph nodes of C57BL/6 mice, and stimulate purified CD4+ T cells under control and Th17-inducing environments. The Th17-inducing environment includes stimulation in the presence of anti-CD3 and anti-CD28 antibodies, IL-6, and TGF-β. After incubation for at least 72 hours and for up to five days at 37 °C, cells are subsequently analyzed for the capability to produce IL-17 through flow cytometry, qPCR, and ELISAs. Th17 differentiated CD4+CD25- T cells can be utilized to further elucidate the role that Th17 cells play in the onset and progression of autoimmunity and host defense. Moreover, Th17 differentiation of CD4+CD25- lymphocytes from distinct murine knockout/disease models can contribute to our understanding of cell fate plasticity.
authors.affiliation: Department of Microbiology and Cell Science, The University of Florida.
authors.forename: Simone K
authors.initials: SK
authors.lastname: Bedoya
authors.forename: Tenisha D
authors.initials: TD
authors.lastname: Wilson
authors.forename: Erin L
authors.initials: EL
authors.lastname: Collins
authors.forename: Kenneth
authors.initials: K
authors.lastname: Lau
authors.forename: Joseph
authors.initials: J
authors.lastname: Larkin
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology; D003584:Cytological Techniques / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D058504:Th17 Cells / Q000166:cytology*