Figure 5. FXII influences DCs in the CNS.

(a) EAE development is shown in WT mice after adoptive transfer of CD4⁺ lymphocytes and CD11c⁺ cells isolated from WT or Cd87^−/− mice on day 12 post immunization and restimulated with 10 μg ml⁻¹ MOG_35–55 and 0.5 ng ml⁻¹ IL-12 with or without 60 nM FXII for 72 h. Mean clinical and mean cumulative scores±s.e.m. over time of three independent experiments are given (non-parametric Mann–Whitney U-test). (b) BM chimeras were created by transferring WT and Cd87^−/− BM into WT and Cd87^−/− recipient mice after radiation. Mean clinical and mean cumulative scores±s.e.m. of EAE from three independent experiments are shown (non-parametric Mann–Whitney U-test). (c,d) Brain-infiltrating leukocytes (BILs) were separated into cDC⁺ and cDC⁻ by sorting via flow cytometry based on indicated surface markers at d_max after EAE induction. Both subsets from WT or F12^–/– mice were analysed for Il-6 expression by real-time reverse transcription–PCR using 18s rRNA for normalization. Data are given as mean±s.e.m. of two experiments, each experiment generated from pooled brain and spinal-cord-derived cells of n=6–7 mice per group and presented as fold change in normalized gene expression relative to WT controls. (e) Flow cytometric analysis of BILs from WT and F12^−/− animals at d_max after EAE induction determined the expression of CD80, CD86 and MHC-II in cDCs that were pre-gated for CD45^highCD11b⁺CD11c⁺ cells. Data are representative of two independent experiments with four mice per genotype. (f) Histological analysis of spinal cord sections stained for the nucleus (4,6-diamidino-2-phenylindole (DAPI), blue), FXII (red) and CD11c (green) from the lumbar region of EAE WT animals at d_max. Scale bar, 100 μm. *P<0.05.