Fig. 5.

Meningeal mast cell-derived caspase-1 is necessary for EAE development and Th cell encephalitogenicity in the CNS. The meninges of naïve Kit^W/Wv mice were selectively reconstituted with wild type (W/W^v + WT MCs) or Casp1^−/−(W/W^v + C^−/− MCs) bone marrow-derived mast cells. (a) Reconstitution was confirmed in a subset of mice by toluidine blue staining of the calvarial dura mater. Arrows denote toluidine blue-positive mast cells. (b) Eight weeks post reconstitution, EAE was induced by MOG_35–55 peptide immunization and the disease course was compared to non-reconstituted controls. *p < 0.05 by 2-way ANOVA. n ≥ 12 for each group. 4 independent experiments. Data points are expressed as mean ± SEM. (c–k) Twelve days post immunization, CNS infiltrating leukocytes were isolated from a subset of the immunized mice and analyzed by flow cytometry. Representative analyses of GM-CSF (c), IL-17 (f), and IFNγ (i) production by CD45^hiCD3⁺CD4⁺ cells are shown. FMO negative controls were used to set gates. Histogram colors correspond to experimental groups designated in (b). (d,g,j) Frequency and (e,h,k) numbers of CD45^hi CD3⁺CD4⁺ cells that express cytokines *p < 0.05, **p < 0.01, and ***p < 0.001 by Student’s t-test. n ≥ 8 for each group, 3 independent experiments.