Fig. 6

CSF1R antagonism increases CNS infiltration by myeloid cells before neurological symptoms appear in EAE. Single cells isolated from pooled brain and spinal cord tissues were analyzed by flow cytometry for myeloid subsets before onset of neurological deficits (preonset) A Numbers of total cells, and of infiltrating immune cells (CD45hi), which include myeloid cells (CD45hiCD11b+) and lymphocytes (CD45hiCD11b-) were increased while microglia were decreased in the CNS of PD compared to CD mice in preclinical EAE. B Gating strategy for flow cytometric analysis of myeloid cells in the CNS. C Frequencies and absolute numbers of neutrophils (CD45hiCD11b+Ly6G+) were increased in the CNS of PD mice with during preonset EAE compared with CD mice. D No differences in the frequencies of total monocytes (CD45hiCD11b+Ly6G-CD11c-MHCII-), total macrophages (CD45hiCD11b+Ly6G-CD11c-MHCII+) and total DCs (CD45hiCD11b+Ly6G-CD11c+MHCII+) were detected but their numbers were increased in the CNS of PD compared to CD. E Frequencies and numbers of cDCs (CD45hiCD11b+Ly6G-CD11c+MHCII+CD26+CD88-), but not of moDCs (CD45hiCD11b+Ly6G-CD11c+MHCII+CD26+CD88+) were increased in the CNS of PD compared to CD mice. F Numbers of Ly6C+ monocytes, and of both Ly6C+ and Ly6C- macrophages were increased in the CNS of PD compared to CD mice in preclinical EAE. Representative flow plots are shown. Both male and female mice were employed. Data are shown as means ± SD, n = 22; *p < 0.05, ***p < 0.0005, ****p < 0.00005