Figure 4. Anti-CCL22 treatment reduced CNS accumulation of Ly6C^hi macrophages.

(A) CNS-infiltrating CD11b⁺Ly6C^hi macrophages from acute EAE express CCR4. Mice were immunized to develop EAE, and at the peak of acute clinical disease, the CNS mononuclear cells were harvested and assessed for CCR4 expression by flow cytometry. The results are shown for a representative animal (out of a total of five), the data are derived from a CD45^hiCD11b⁺ gate, and the results show CCR4 expression as a function of Ly6C^hi expression. (B) CNS-infiltrating CD11b⁺Ly6C^hi macrophages from relapsing EAE express CCR4. Mice were immunized to develop EAE, and at the peak of relapsing clinical disease, the CNS mononuclear cells were harvested and assessed for CCR4 expression by flow cytometry. The results are shown for a representative animal (out of the remaining five), the data are derived from a CD45^hiCD11b⁺ gate, and the results show CCR4 expression as a function of Ly6C^hi expression. (C) The presence of Ly6C^hiCD11b⁺ macrophages was assessed in the CNS of control Ig-treated mice at peak clinical disease using flow cytometry. The results are displayed from a representative animal (out of a total of three) as the percentage of total CD11b⁺ macrophages out of the CD45⁺ gate (R1=37%) and the percentage of only Ly6C^hi cells from the CD45⁺ gate (R2=18%). (D) The presence of Ly6C^hiCD11b⁺ macrophages was assessed in the CNS of anti-CCL22-treated mice at the time controls were showing peak clinical disease using flow cytometry. The results are displayed from a representative animal (out of a total of three) as the percentage of total CD11b⁺ macrophages out of the CD45⁺ gate (R3=22%) and the percentage of only Ly6C^hi cells from the CD45⁺ gate (R4=5%). The results are representative of two similar experimental replicates.