Figure 5
IFN-γ, IL-10 and nitrite production by splenocytes from control, EAE and treated animals. Splenocytes (8 × 105 cells per well) were obtained from Control, EAE, and EAE + NAC treated rats when they were showing maximum clinical signs, and were stimulated in vitro with PHA (10 μg/ml, a & b), MBP (20 μg/ml, a & b) or LPS (1 μg/ml, c) for 48 hrs. Each treatment was performed in triplicate for each animal group (n = 6). a and b: The levels (pg/ml) of IFN-γ and IL-10 in culture supernatants were measured using ELISA kits. There was a significant increase in IFN-γ and decrease in IL-10 in splenocytes from untreated EAE animals stimulated with PHA and MBP. EAE + NAC splenocytes showed reduced IFN-γ production whereas IL-10 production was increased. c: Culture supernatants were collected and accumulated nitrite, a stable product of NO production, was measured using Griess reagent. LPS-stimulated EAE splenocytes showed significantly higher levels of nitrite as compared to control and this was reduced with NAC treatment.