Fig. 8

Effect of the local activation of highly polarized Th1, Th2, and Th17 CD4⁺ T cells on the corneal epithelium and nerves of T cell-reconstituted mice. T cell-deficient mice were reconstituted with in vitro polarized CD4⁺ T cells. Transferred mice were later given OVA or saline (PBS) eye drops daily for 4 weeks as detailed in the previous figure. A group of non-transferred (–) littermates was also included. A Representative micrographs of corneal whole mounts stained with E-cadherin (red) and tubulin β3 (green) from immunized wt mice. B Representative micrographs and C pooled data of corneal dextran-fluorescein uptake in transferred mice. D Corneal mechanical sensitivity thresholds of reconstituted mice given OVA eye drops over 4 weeks. The dotted reference line corresponds to the average baseline measurements of all the mice in the experiment. E Schematic of the levels at which nerve morphology was analyzed in corneal whole mounts stained with tubulin β3 (green). F Quantification (cumulative data) of intraepithelial nerve terminals imaged en face beneath the apical epithelial squamous cells (subapical section) and in cross-section at the mid-epithelial level (count of nerve endings/field) as they run perpendicularly to the surface, and of corneal neural complexity at the subbasal level (sum of intersections, Sholl analysis). G Representative micrographs of corneal intraepithelial nerves at the three different levels. 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 was used in B and C (treatment and time) and one-way ANOVA with Dunnett's post hoc test was used in (F). *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant