Inhibition of Corneal Inflammation by the TLR4 Antagonist Eritoran Tetrasodium (E5564)

PURPOSE. To investigate the role of the TLR4/MD-2 antagonist eritoran tetrasodium in a murine model of contact lens-associated corneal infiltrates. METHODS. C57BL/6 mouse corneas were abraded and treated with eritoran tetrasodium or placebo, either before or after stimulation with either LPS, the TLR2 ligand Pam(3)Cys, or antibiotic-killed Pseudomonas aeruginosa. A 2-mm punch from a silicon hydrogel contact lens was used to cover the corneal surface throughout the inhibition and stimulation period. Corneal infiltrates were detected by in vivo confocal microscopy and by immunohistochemistry for neutrophils. The effect of eritoran tetrasodium on stimulated human corneal epithelial cells (HCECs), macrophages, and neutrophils was also assessed. RESULTS. Eritoran tetrasodium significantly inhibited CXC chemokine production in the cornea and development of corneal infiltrates, specifically neutrophils, in response to stimulation with LPS (TLR4), but not Pam(3)Cys (TLR2). When the antagonist was applied after LPS stimulation, neutrophil infiltration was also inhibited, although a higher concentration was needed. Furthermore, IL-8 production by TLR4- but not TLR2-stimulated HCECs, macrophages and neutrophils was also significantly reduced. Corneal inflammation induced by P. aeruginosa in the presence of tobramycin was found to be dependent on expression of TLR4 and MD-2 and is inhibited by eritoran tetrasodium. CONCLUSIONS. Eritoran tetrasodium is a highly effective antagonist of TLR4/MD-2-dependent corneal inflammation. (Invest Ophthalmol Vis Sci. 2009; 50: 1247-1254) DOI:10.1167/iovs.08-2628