PURPOSE. The purpose of this study was to investigate the effectiveness of tumor necrosis factor (TNF)-a blocker for treatment of dry eye (DE)-induced inflammation and determine a more effective method to suppress lacrimal gland inflammation. Efficacy of topical versus systemic administration of TNF-a blockers was determined using a murine dry eye (DE) model. METHODS.The TNF-a blocker HL036 was developed by modification of the TNF receptor I. Protein purity, binding affinity, and clearance of TNF-a was compared with etanercept. Using DE-induced C57BL/6 mice, corneal erosion and goblet cell counts were measured after subcutaneous or topical treatment with etanercept or HL036. Inflammatory cytokines in cornea and lacrimal glands were determined by quantitative RT-PCR and ELISA.RESULTS. HL036 showed TNF-a binding affinity comparable to etanercept, as measured by surface plasmon resonance. HL036 concentration was significantly higher in cornea and anterior segment than etanercept and effectively eliminated TNF-a on ocular surfaces. Etanercept was preferentially concentrated in the posterior segment. Corneal erosion and goblet cell counts were improved only with topically applied etanercept and HL036. Ocular surface IFN-c, IL-6, and IL-21 were significantly decreased by topical HL036. DE-induced lacrimal gland IFN-c and IL-6 expression was effectively suppressed by topical etanercept and HL036.CONCLUSIONS. Topical TNF-a blockers effectively suppressed lacrimal gland and corneal inflammation by suppressing IFN-c, IL-21, and IL-6. Differences in TNF-a affinity, clearance, and local concentration of blockers may account for the anti-inflammatory effects in different ocular regions.

Although corneal allotransplantation is performed in the immune-privileged cornea, many grafts are still rejected after transplantation. This study examined the role of chemokine receptor D6 expression in a corneal allograft rejection, investigated the modulation of D6 expression in cells, and determined the effect of D6 on graft survival. Interestingly, D6 was highly expressed in CD45 -: cells and the corneal epithelium of accepted corneal allografts. From the mouse corneal allograft model, TGF-β was found to play a key role in D6 up-regulation, leading to reduced CCL2, CCL5, and CCL3. To modulate D6 chemokine binding, a D6MT was developed and showed effective chemokine trapping through SPR and FACS assays. By treating corneal allografts with D6MT, the allograft survival rate was improved, and (lymph) angiogenesis was reduced. Direct allosensitization and DC LN homing was drastically reduced in the mouse corneal allograft model. These findings suggest that TGF-β is a positive regulator of D6 expression, and it is a potential therapeutic target to enhance the survival of corneal allografts.