Fig. 5

TRPV1 signaling in the injured eye and SP release in the contralateral eye mediate sympathetic ocular mucosal tolerance disruption after a unilateral corneal burn. a Experimental design for dissecting the inter-eye neurogenic inflammatory reflex contribution to the contralateral ocular mucosal response to ovalbumin (OVA) in the context of a corneal burn. On day 1, either of two TRPV1 blockers (SB-366791 and BCTC, respectively) were administered repeatedly to the right eye (OD) and/or a substance P blocker (aNK1R) was repeatedly applied on the left eye (OS) of mice 1 h before inducing a corneal burn in the right eye. Antigen (OVA) was instilled on the left eye from days 2 to 5, and then s.c. immunization (Imm) was performed with complete Freund’s adjuvant. The antigen-specific immune response was measured by footpad swelling in a delayed-type hypersensitivity (DTH) challenge. b Antigen-specific swelling measured 48 h after s.c. OVA injection and expressed as the difference with the saline-injected contralateral footpad. Pooled data (mean ± SD) from 4 independent experiments with at least 3 mice/group (two-way ANOVA with Bonferroni’s correction). c Dot plot (left) and representative confocal micrographs (right) of nuclear localization of NF-κB p65 protein (green) relative to individual cell expression (blue: TO-PRO-3, red: phalloidin) in the conjunctival epithelium of mice from experiment a. Both eyes from each mouse (burnt and opposite) were harvested 48 h after initiation of the experiment and analyzed separately. Mean ± SD of at least 100 cells analyzed from samples from 3 mice/group (representative example of 3 independent experiments), analyzed by one-way ANOVA with Bonferroni’s correction. For all panels, asterisk (*) indicates a statistically significant difference by the corresponding test