Reduced splenic expression of genes required for macrophage response to pathogens. (A) Analysis of toll-like receptor (TLR) transcripts show downregulated expression of the majority of tlr genes 1–5 days after middle cerebral artery occlusion (MCAO) in comparison to sham-operated animals (sham n = 2, 1 day and 2 days n = 3, 5 days n = 4). (B) RT-qPCR for Tlr1 confirmed significant downregulation at 2–5 days after MCAO in comparison to sham-operated animals (naïve n = 4, sham n = 4, 1 day n = 4, 2 days n = 9, 5 days n = 5, 7 days n = 7). (C) Expression of transcripts which encode macrophage-associated co-stimulatory proteins important for activating the adaptive immune response show downregulated expression 1–5 days after MCAO in comparison to sham-operated animals (sham n = 2, 1 day and 2 days n = 3, 5 days n = 4). (D) RT-qPCR for Cd40 confirmed significant downregulation at 2–5 days after MCAO in comparison to sham-operated animals (naïve n = 4, sham n = 4, 1 day n = 4, 2 days n = 9, 5 days n = 5, 7 days n = 7). (E) Expression of Tnfa which encodes the cytokine TNFα was downregulated 1–5 days after experimental stroke whereas expression of Il-10 which encodes the cytokine IL-10 remained relatively unchanged (sham n = 2, 1 day and 2 days n = 3, 5 days n = 4). (F) RT-qPCR for Tnfa confirmed downregulation of expression 1–7 days after MCAO in comparison to sham-operated animals (naïve n = 4, sham n = 4, 1 day n = 4, 2 days n = 9, 5 days n = 5, 7 days n = 7). (G) CBA assay of homogenized spleen detected TNFα in the spleens of sham-operated animals and animals recovered 7 days after MCAO; however, this was undetectable in spleens from mice recovered 1–5 days after MCAO. Data show mean ± SD (B,D,F), *P < 0.05; **P < 0.01; one-way analysis of variance with Bonferonni correction.