Eosinophils do not contribute to EAE. (A) Flow cytometry overlay of BM-derived eosinophil (Eos) cultures showing double-staining for Siglec-F and CCR3 (blue) and with isotype monoclonal antibodies (red). (B) Adoptive transfer of BM-derived eosinophils. WT and ΔdblGATA mice were immunized for EAE, and 2 × 10⁷ eosinophils were injected IV on days 7 and 9 after immunization into ΔdblGATA mice. (C) Clinical scores and change in body weight are shown. Representative figures from 1 of 2 independent experiments. Significance was calculated by 2-way ANOVA (n = 4 for WT and ΔdblGATA groups; n = 6 for ΔdblGATA and eosinophil group). (D) Clinical course of EAE in WT C57BL6/J and PHIL mice (n = 5 per group). (E) Relative numbers of CD45⁺ cells isolated from the CNS (brains and spinal cords combined) of WT and PHIL mice. (F) Frequencies of CD11b⁺ and CD4⁺ cells within CD45⁺ cells from the CNS of WT and PHIL mice. (G) Frequencies of Th1 (interferon γ⁺ [IFN-γ⁺] IL-17A), Th17 (IL-17A⁺IFN-γ⁻), Th1/17 (IFN-γ⁺IL-17A⁺), regulatory T cell (Treg) (FoxP3⁺), and GM-CSF⁺ T helper cells from the CNS of WT and PHIL mice. (H) Proliferation assay. Mononuclear cells isolated from the CNS of WT and PHIL mice were stimulated for 18 hours with 25 μg/mL MOG_35-55 (immunizing peptide) and 1 μCurie per well of [³H]adenine was added for additional 18 hours. Cells were then harvested, and counts per minute (C.P.M.) were measured using beta counter. (I) Clinical scores for adoptive EAE in WT, PHIL, 1638 (IL-5tg), and 1638/PHIL mice. Abs, antibodies; Freq., frequency.