Fig. 4

Effect of a local Th1- and Th2-skewed immune response on the corneal epithelium and nerves of wild-type mice. Wild-type (wt) mice were immunized with ovalbumin (OVA) in combination with complete Freund’s adjuvant (CFA) or alum and later given OVA or saline (PBS) eye drops, as detailed in the previous figure. A Representative micrographs of corneal whole mounts stained with E-cadherin (red) and tubulin β3 (green) from immunized wt mice. B Cumulative data and C representative of corneal dextran-fluorescein uptake in immunized wt mice. The dotted reference line corresponds to wt mice with dry eye surgically induced on the same day as the ocular challenge (positive control for corneal epitheliopathy). D Corneal mechanical sensitivity thresholds of immunized mice over the 31-day experiment. The dotted reference line corresponds to the average baseline measurements of all the mice in the experiment. E Schematic of the levels at which corneal nerve morphology was analyzed. F Density of vertical intraepithelial nerve terminals in mid-epithelial corneal sections (representative example). G Representative micrographs of mid-peripheral subbasal sections (tubulin β3 staining) and H pooled data (representative experiment) of corneal neural complexity quantification (sum of intersections, Sholl analysis). All experiments were performed twice or more with 6 mice/group/experiment. For all experiments, mean ± standard error of measurement is shown. To compare means, two-way ANOVA (immunization and ocular challenge) was used. *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant