Fig. 6

Effect of a local Th1- and Th2-skewed immune response on the corneal epithelium and nerves of CD4⁺ T cell-reconstituted mice. T cell-deficient mice were reconstituted with CD4⁺ T cells from mice immunized with ovalbumin (OVA) in combination with complete Freund’s adjuvant (CFA) or alum. Transferred mice were later given OVA or saline (PBS) eye drops daily for 4 weeks as detailed in the previous figure. A Representative micrographs and B pooled data of corneal dextran-fluorescein uptake in reconstituted mice challenged with ocular OVA. C Corneal mechanical sensitivity thresholds of reconstituted mice given either PBS or OVA eye drops over 4 weeks. The dotted reference line corresponds to the average baseline measurements of all the mice in the experiment. D Schematic of the levels at which nerve morphology was analyzed in corneal whole mounts stained with tubulin β3 (green). E 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). F 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 A and C (treatment and time), and E (immunization and challenge). *p < 0.05, **p < 0.01, ***p < 0.001, and ns not significant