Adenovirus type 37 keratitis in the C57BL/6J mouse

Abstract

Purpose: To develop a mouse model of adenoviral keratitis that will allow further study of viral and host pathogenic mechanisms.

Methods: Corneas of C57BL/6J mice were injected with adenovirus type 37 (Ad37) or virus-free dialysis buffer by a gas-powered microinjection system coupled to a glass micropipette needle. Mouse corneas were examined for signs of inflammation, by clinical examination, immunohistochemistry, and confocal microscopy; assayed for viral and chemokine mRNA expression by real-time PCR; titered to assess viral replication; and subjected to ELISA for chemokine and myeloperoxidase (MPO) protein expression.

Results: C57BL/6J mice corneas injected with 10(5) TCID (tissue culture infective dose) Ad37 showed stromal opacification and inflammation beginning from 1 day after injection and continuing for several months, while buffer-injected corneas showed no signs of inflammation. Ad37-injected corneas expressed adenoviral E1A 10S and E1B 19k mRNA but not IIIa, and viral titers had fallen two logs by day 4 after injection. When compared to untouched and buffer-injected corneas, Ad37-injected corneas expressed significantly higher levels of IL-6, KC, and MCP-1 mRNA at 4 hours after injection (P < 0.05). By ELISA, KC protein was significantly elevated in Ad37-injected corneas at 8 and 16 hours, and MCP-1 protein at 16 hours after injection (P < 0.05). Ad37-injected corneas showed elevated levels of MPO (P = 0.0024) at 4 days after injection consistent with immunohistochemical evidence for a predominance of neutrophils in the corneal stroma.

Conclusions: Ad37 induces an acute immunopathologic response in the C57BL/6J mouse cornea, despite an absence of viral replication. This new animal model of Ad37 keratitis will facilitate studies of the molecular pathogenesis of the disorder.