Fig. 4

PD-1 agonism prevents the progression of EAE. A EAE was induced in WT C57Bl/6 J mice. Clinical course was blindly monitored. One day after peak disease, a PD-1 agonist (n = 6) or vehicle control (n = 6) treatment was randomly assigned and injected i.p. for 5 consecutive days. Data are representative of two independent experiments. B–M 25 days post-immunization, mice were sacrificed and the CNS was collected and cryopreserved for IHC analysis. Ventral white matter tracts of the lumbar spinal cord were imaged using confocal microscopy. Spinal cord tissue from (B) vehicle- and (C) PD-1 agonist-treated mice were labeled for GFAP, MBP, CD45, and nuclei were counterstained with DAPI. Tissues from (D, F) vehicle- and (E, G) PD-1 agonist-treated mice were also labeled for CD11b, pSHP2, CD45, and nuclei were counterstained with DAPI. Total (H) GFAP, (I) MBP, (J) lesion area, (K) CD11b, and (L) pSHP2 were quantified using ImageJ. M Colocalization of CD11b with pSHP2 was also assessed. Data represent the mean ± SEM combined from 2 independent experiments and were analyzed using the (A) Mann–Whitney U test for nonparametric data or by (H–M) two-tailed Student’s t test. *P < 0.05, **P < 0.01, ****P < 0.0001