Fig. 2. Effects of rIL-10 and B cells on microglial activation.

Primary MG, isolated and cultured from naïve WT male mice, were harvested after 21 days in vitro (at confluency) and cultured in GM-CSF-free medium for 5 days. MG were stimulated with 10ng/ml LPS for 4 hours. Supernatants were discarded after 4 hours and one of the following treatments was given in 1mL of fresh culture medium: no treatment, 20ng recombinant IL-10 (rIL-10), IL-10⁺ B cells at a 1:1 ratio or at a 1:2 ratio with MG. The MG cells were incubated with mentioned treatments at 37°C and 5% CO₂ for 24 hours and proinflammatory cytokines A. TNF-α and B. IL-β and C. chemokine CCL3 (MIP-1α) were determined by flow cytometry. Values are given as mean ± S.E.M. Data presented are representative of n = 5, 3 and 2 separate co-culture set-ups, respectively for TNF-α, IL-1β and CCL3 with each condition done in duplicate for every experimental set-up. Statistical analysis was performed using One way ANOVA with post-hoc Dunnett test. Significant differences between sample means are indicated as * p≤0.05, ** p≤0.01, ***p≤0.001 as compared to the LPS-stimulated condition