TIMP-1 role in protection against Pseudomonas aeruginosa-induced corneal destruction

To establish the role of TIMP-1 in protection against Pseudomonas aeruginosa-induced corneal destruction, corneas of adult 8-week-old (resistant) and aged 12-month-old (susceptible) mice were infected with the bacterium. Corneas were analyzed for TIMP-1 protein by immunocytochemistry and Western blotting. Basement membrane (BM) integrity was assessed by immunostaining for type IV collagen. Additionally, resistant 8-week-old mice were treated systemically with neutralizing TIMP-1 polyclonal antibody (pAb) or pre-immune normal rabbit serum (NRS). Ocular and BM integrity as well as MMP-9 expression were examined in these mice. A greater amount of TIMP-1 protein was observed in the cornea of 8-week-old mice. In the cornea, the strongest staining was found in the superficial epithelium, but positive staining also was seen in the basal epithelium and stroma. When type IV collagen was analyzed in the BM of both age groups of mice, a distinct staining pattern was observed in only the young adult mice. Treatment of 8-week-old resistant mice with neutralizing TIMP-1 pAb vs NRS increased the amount of MMP-9 in the cornea of TIMP-1 pAb-treated mice and affected the ability of these mice to deposit BM components. These studies suggest that adequate expression of TIMP-1 protects against BM and stromal degradation via multiple processes.