Figure 2. ICOS is not crucial for TH-17 differentiation in vivo nor for induction of acute EAE.
(a-e) ICOS-deficient (Icos-/-) or wild-type mice were immunized with MOG35-55 emulsified in complete Freund's adjuvant to induce EAE. (a) Flow cytometry for intracellular cytokine secretion in CD4+ T cells at day 10 obtained from mononuclear cells isolated from CNS, dLN and spleen activated with PMA-ionomycin. (b-d) Lymph node cells from immunized mice were tested for antigen-specific response: proliferation in triplicate cultures by 3[H] thymidine incorporation (mean ± s.d.) (b), cytokine production by ELISA (day 2) (c) and intracellular cytokine staining by flow cytometry (day 5) (d). (e) The mean clinical EAE scores for the ICOS-deficient (dark circles) and wild-type control (open circles) groups (mean ± s.e.m.). Numbers adjacent to boxed areas or in quadrants indicate percent positive cells in each. Data represent one of four experiments (a) or the mean ± s.d. of five (b,c,d) or four (e) experiments.