FIG 4.

Impact of RAG-1 deficiency on cytokine and chemokine mRNA levels 24 h after induction of meningitis in wild-type C57BL/6 mice and RAG-1-deficient mice. Quantitative real-time PCR was performed using hypoxanthinguanine-phosphoribosyltransferase (A) or beta-actin (B to G) as housekeeping genes. (A) Expression of interleukin-10 (IL-10), gamma interferon (IFN-γ), transforming growth factor β (TGF-β), forkhead box P3 (Foxp3), and tumor necrosis factor alpha-related ligand (TRAIL) mRNAs in cerebellar tissue lysates. Regulation is shown relative to nonstimulated controls of the same genotype using the ΔΔ_CT method. (B to D) Activated microglia/macrophages were isolated from cerebral tissue as CD11b⁺ CD45^high cells using FACS and studied for the expression of IL-10 mRNA (B), as well as for the regulation of TGF-β mRNA (C) and CCL2 mRNA (D). (E to G) CD11b⁺ myeloid cells were isolated from the CSF and studied for the regulation of IL-10 mRNA (E), CCL2 mRNA (F), and CXCL1 mRNA (G) relative to controls. Graphs depict mean values and standard deviations. Statistical significance was examined using Student t tests. The detection limit was set at 40 PCR cycles. In panels B and G, negative results are marked with an “n.d.” (not detected), and expression levels are shown as 1/Δ_CT.