Enhanced neurotoxoplasmosis. Immunofluorescence staining of (A) F4/80, (B) CD8α and (C) CD3+ cells in the brains of MyD88−/− and MyD88+/+ mice. Frozen brain sections were stained with rat anti-F4/80, rat anti-CD8 and rat anti-CD8, followed by an Alexa 594-conjugated secondary antibody against rat IgG. Sections were counterstained with Hoechst (x 100). (D) Expression of chemokine mRNA in the brains of MyD88+/+ (open bars) and MyD88−/− (solid bars) BALB/c mice 13 days postinfection evaluated by qRT-PCR. Results are presented as the fold increase expression of mRNA in infected mice versus expression of mRNA in non-infected mice. The results are representative of two different experiments. The total number of tested animals in each group is n = 7. (E) Determination of cytokine levels in supernatants of brain homogenates from MyD88+/+ and MyD88−/− BALB/c mice 13 days postinfection. Supernatants from MyD88+/+ (open bars) and MyD88−/− (solid bars) were collected to determine cytokine levels. Results are representative of two experiments and were analyzed using a Mann–Whitney test. * (P <0.05). qRT-PCR, real-time reverse-transcription polymerase chain reaction.