Fig. 6
GITRL-competitive mouse GITR blocking antibody is protective in an imiquimod skin inflammation model. (A) Schematic of Imiquimod model. 5% imiquimod cream was applied on the left and right ear of 10–12 weeks old female C57Bl/6 mice for 5 days. Mice were dosed i.p. on days − 2 and day 2 with anti-mouse GITR antibody (2 mg/kg, or 10 mg/kg), or isotype control (10 mg/kg), and on day 0 and day 3 with anti-mouse IL17A antibody (20 mg/kg). Ear thickness was measured daily (days 0–5) in triplicate. Mice were sacrificed on day 5. Ear and spleen samples were collected for flow cytometry analysis. (B) Mice treated with muGITR-lc-Ab-#5 at 2mpk and 10mpk and IL-17 A antibody showed significantly less ear swelling compared to isotype control; day 5 measurements. (C) Mice treated with muGITR-lc-Ab-#5 at 10mpk and IL-17 A antibody had less CD11b + monocyte in the ear compared to isotype control. (D) Mice treated with muGITR-lc-Ab-#5 at 2mpk and IL-17 A antibody showed significant lower CTLA4 MFI in Foxp3- CD4 effector T cells in spleen. (E) Mice treated with muGITR-lc-Ab-#5 at 2mpk and 10mpk but not IL-17 A antibody showed significant lower % macrophage in CD11b + cells in spleen. (F) Mice treated with muGITR-lc-Ab-#5 at 2mpk and 10mpk and IL-17 A antibody showed significant lower % CD4 cells in CD3 in the spleen. Horizontal bar represents average of the group. Data were compared to isotype control with Å Ãdák’s multiple comparisons test after one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. m mouse, mpk mg per kilogram, i.p. intraperitoneal, lc ligand competitive, d day.
