Fig. 2.

Treatment with Ind-Cl suppresses cytokine production by peripheral immune cells and reduces CNS inflammation and infiltration. (A) Cytokine production by MOG_35–55-stimulated splenocytes was assessed from EAE mice killed on postinduction day 34. Pretreatment with DPN or Ind-Cl was initiated at day 0, and posttreatment with Ind-Cl began on day 21. DPN-treated (blue) and vehicle-treated (red) animals displayed similar cytokine levels. Pretreatment (light purple) with 5 mg/kg/d Ind-Cl resulted in significant reduction of all measured cytokines. Posttreatment with Ind-Cl (dark purple) significantly decreased TNFα levels. Data are representative of experiments repeated twice. n = 4–6 mice/group; *P < 0.05, t test. (B) CNS inflammation was assessed using immunohistochemistry. Asterisks within the representative 4× image of vehicle-treated EAE spinal cord indicate lesions and areas of infiltration and demyelination, unlike Ind-Cl-treated mice (i and ii). Ten times and 40× images of the DC (area delineated by the white dashed box) in i shows decreased infiltration by peripheral CD45⁺ immune cells with Ind-Cl treatment (ii and iii) and decreased CD3⁺ T-cell (red) numbers (iv). Pretreatment with Ind-Cl resulted in decreased GFAP⁺ (red) intensity and GS+ (red) numbers, and a trend toward this effect was observed with posttreatment (v and vi). n = 10 mice/group; *P < 0.05, **P < 0.01, ANOVA.