Figure 5

Peripheral and central immune cell phenotypes are not altered following anti-TNFR1 treatment. MNCs, isolated from the CNS (pooled brain and spinal cord; (A), and splenocytes (C) were collected on days 5 and 6 of acute EAE from mice treated with either ATROSAB or control IgG. Following gating for CD45⁺ CD4⁺ cells, FACS analysis was performed to determine the percentage of those co-expressing markers of either TH1 (IFNγ), TH17 (IL-17), or Tregs (FoxP3). In addition, following gating for CD45⁺ cells from the CNS (B) and spleen (D) on days 5 and 6 of acute EAE, cells were assessed for markers of monocytes (CD11b). No differences could be seen between cells from the two treatment groups (n = 4 mice per group). In addition, immunohistochemistry was performed to determine the percentage of CD3⁺ spinal cord infiltrates (black) co-expressing FoxP3 (brown), in order to identify Tregs. In comparison to control-treated animals, no difference was seen in the percentage of CD3⁺FoxP3⁺ cells in the ATROSAB-treated groups at either the acute (F–G) or chronic (H–J) disease stages. n = 6 per group. ATR, ATROSAB; CON, control IgG. Scale bar = 200 µm.