Figure 8

Differences in T cell activation translate into differences in IFN-γ production, which affects the microglial expression of complement factors and complement inhibitors. (A) Normalized relative expression of IFN-γ mRNA in relation to the house-keeping gene beta actin (calculated using the following equation: 2^-ΔCt = 2^{-[Ct(GOI)-Ct(HKG)]} (GOI – Gene of interest; HKG – house-keeping gene; [19]) are shown. Statistically significant differences (*, as determined by one-way ANOVA followed by Bonferroni’s post-hoc testing) were observed between MBP-specific T cells and their MOG- or S100β-specific counterparts. (B) Pathways contributing to the complement cascade [20] and alterations in gene expression (encircled in red: upregulation; encircled in green: downregulation) of complement factors and inhibitors by IFN-γ treated microglia. (C-D) Changes in gene expression of complement factors and inhibitors (C) and of Fcgr3 (D) in IFN-γ treated microglia. These cells were treated for 48 hrs with 100 ng/ml IFN-γ. Subsequently, the mRNA of these cells was harvested and subjected to gene expression profiling. Log2-fold changes in gene expression and differences in the normalized signal intensities (nSI) of complement components/factors and Fcgr3 between IFN-γ and vehicle control-treated microglial cultures are shown (2 different, independent samples per treatment group). Genes with elevated expression in the IFN-γ treated group are labeled red, genes with lower expression levels are labeled green.