Figure 5.

Tuftsin, alone and together with benztropine, dampens inflammation in EAE. The lumbar region of the spinal cord was isolated from control or treated mice at day 21 and 28 post-MOG immunization. Iba1 immunostaining (green) was performed to identify microglia/macrophages in the ventral white matter. To identify whether the microglia/macrophage cells were pro-inflammatory, iNOS (red) immunoreactivity was examined and to identify anti-inflammatory cells, Arg1 (red) was used. Nuclei were identified using DAPI (blue) (A). Number of iNOS⁺Iba1⁺ or Arg1⁺Iba1⁺ cells were counted and divided by the total number of Iba1⁺ cells in the field to calculate a percentage (B,C). Pro- to anti-inflammatory ratios were calculated by dividing the percent of iNOS1+ microglia by the percent of Arg1⁺ microglia (D). Linear regression and comparison of slopes was performed on the day 21 and 28 averages to examine temporal regulation of microglia/macrophage polarization by the treatment groups (E,F). Data are mean ± SEM. n = 4–6, *p < 0.05, **p < 0.01. ***p < 0.001 compared to control. Scale bar: 20 um.