Figure 6: FXII blockade protects from neuroinflammation and contact hypersensitivity.

(a) Clinical scores of WT mice treated with rHA-Infestin-4 or vehicle starting on day 1 after EAE induction are shown. Representative data from two independent experiments are depicted (Supplementary Table 3). (b,c) Histological analysis of spinal cord sections of WT and rHA-Infestin-4-treated mice at d_max. Lumbar sections were stained with haematoxylin and eosin (HE) (b) to evaluate inflammatory foci or immunostained for Luxol fast blue (LFB) (c) to assess demyelination. Arrows indicate inflammation or demyelinated areas. Scale bars, 100 μm. (d) Cytokines produced by CD4⁺ T lymphocytes and DCs from WT mice treated with rHA-Infestin-4 or vehicle 10 days after immunization. Data from three independent experiments, each performed in duplicate are shown. Clinical scores of (e) MOG_35–55-immunized WT mice and (f) proteolipid protein peptide 139–151-immunized SJL/JRj mice treated with rHA-Infestin-4 or vehicle starting at the first day of neurologic symptoms (arrow; for detailed information, see Supplementary Table 4). (g) Clinical, cumulative EAE scores and (h) disease incidence of Devic mice treated with rHA-Infestin or vehicle starting at postnatal day 20. (i) The motor coordination of Devic mice was assessed using the rotarod. The time the mice remained on the rod was recorded. For each mouse, the average time of three trials followed by 30-min breaks was recorded daily. (j) Number of infiltrating CD4⁺ T cells within the central nervous system of Devic mice was analysed by flow cytometry at postnatal day 40. (k) Ear swelling of WT or F12^−/− mice following induction of contact hypersensitivity. Representative data from three independent experiments are shown. Quantification of 2,4-dinitrobenzenesulfonic acid sodium salt-induced proliferation, and IFN-γ and IL-17A production of LN cells from WT and F12^−/− mice. Representative data in quadruplicate wells from two independent experiments are shown. In a–j, 200 mg kg⁻¹ body weight of rHA-Infestin-4 or vehicle was given once daily. In a–k, data are given as mean±s.e.m. *P<0.05, **P<0.01, ***P<0.001 (for a,d and e–k, non-parametric Mann–Whitney U-test; for b and c, Student’s t-test); NS, not significant.