Figure 3Figure 3
Microglial activation and motor impairment by AQP4-IgG requires CNS-infiltrating neutrophils. ( A ) Timeline for injecting neutrophil-depleting anti-Ly6G-IgG or isotype control-IgG (100 mg/kg, i.p.), inserting lumbar subarachnoid catheter, infusing AQP4-IgG, and rotarod testing. ( B ) Flow cytometry confirms neutrophil ablation efficiency (percentage CD45 + CD11b + Gr1 + MPO + cells among peripheral CD45 + CD11b + cells). ( C ) (Upper) t-SNE analysis of CD45 + immune cell subtypes from lumbar spinal cords of control and neutrophil-depleted mice. (Lower) Quantification of data in B (3 mice/group). ( D ) Representative confocal images of neutrophils in lungs of mice receiving neutrophil-depleting anti-Ly6G-IgG or isotype control-IgG (3 mice/group). ( E ) Microglial activation, reflected by Cx3cr1GFP signal, in corresponding lumbar cord regions of mice without and with neutrophil ablation (by Ly6G-IgG or isotype control-IgG) after 3 days’ infusion with normal control mouse IgG or AQP