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. Author manuscript; available in PMC: 2006 Oct 22.
Published in final edited form as: Nat Immunol. 2005 Oct 2;6(11):1133–1141. doi: 10.1038/ni1261

Figure 1.

Figure 1

The generation of IL-17-producing T cells requires CD28 and ICOS costimulation. (a,b) B6 mice were immunized with MOG(35–55) in CFA or CFA alone; 7 d later, lymph node and spleen cells from immunized mice (two mice in each group) were collected. (a) ELISA of cytokines in supernatants of lymph node and spleen cells from nonimmunized (Nonimm) and immunized mice stimulated in triplicate with MOG peptide for 72 h in vitro. (b) Intracellular staining for IL-17, IFN-γ or TNF after MOG restimulation. Data are of gated CD4+ cells; numbers beside outlined areas indicate percent positive cells in that area. (c) Intracellular detection of IL-17 and IFN-γ in conjunction with anti-CD4 or anti-CD8. B6 mice were immunized for 7 d with KLH in CFA and then spleen and lymph node cells were collected and restimulated with PMA and ionomycin. Numbers beside outlined areas indicate percent positive cells in that area. (d) ELISA of cytokine production by CD4 T cells isolated from Icos-sufficient (Icos+/+) or Icos-deficient (Icos−/−) OTII mice and activated for 4 d with splenic APCs from B6 mice (B7 WT) or mice doubly deficient in B7.1 and B7.2 (B7 KO) in the presence of OVA peptide; activated cells were then washed and restimulated for 24 h with anti-CD3. (e) ELISA of cytokine production by naive T cells isolated from Icos+/1 and Icos−/−mice with or without the Maf transgene and activated for 4 d with anti-CD3, APCs and IL-2. (f) ELISA of cytokine production by spleen cells isolated from B6, B7-deficient (B7 KO), Icos−/− or B7-deficient Icos−/− mice (three in each group, analyzed individually) immunized with KLH in CFA; spleen cells were restimulated ex vivo with various doses of KLH. Data are representative of at least two independent experiments with similar results.