Fig. 1.

Intrathecal MIS416 is taken up by phagocytic monocytes and neutrophils in CNS. a Representative flow cytometry profile showing the distribution of CD45^high and CD45^dim cell populations in the CNS of control (Ctrl) and MIS416-treated mice at 2, 4 and 24 h. MIS416⁺CD45^high cells were detected in the CNS following intrathecal MIS416 delivery. The MIS416⁺CD11b^highCD45^high cell population included F4/80⁺GR-1^lowCD11c^−/+ (a and c and d) and F4/80⁻Gr-1^highCD11c⁻ cells (b and c). b bar graphs show proportion of CD45^high cells at 2, 4 and 24 h. c Representative flow cytometry profile showing GR-1⁺MIS416⁺ cells to be Ly6c^lowLy6G^high. d Micrographs showing colocalization of MIS416 (green) with CD45⁺ or Ly6G⁺ (red) cells (arrows) located in the extraparenchymal compartment of the CNS that had received MIS416-AF88 by intrathecal injection 4 h before. DAPI (blue) shows nuclear staining. e MIS416⁺GR-1^high and MIS416⁺F4/80⁺ cells were analyzed for the expression of PDL-1 by flow cytometry. Gray histograms represent fluorescence minus one (FMO) staining, and black histograms represent positive staining for PDL-1. Data are presented as mean ± SEM. (n = 5–12 per group). Results were analyzed using the two-tailed Mann-Whitney u-test; * p < 0.05, ** p < 0.01, *** p < 0.001