Adenovirus type 37 keratitis in the C57BL/6J mouse.
OBJECTIVE
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.
Chintakuntlawar
Ashish V
AV
Molecular Pathogenesis of Eye Infection Research Center, Dean A. McGee Eye Institute--OUHSC, Oklahoma City, OK 73104, USA.
Astley
Roger
R
Chodosh
James
J
D000257:Adenoviridae Infections / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D017500:Adenovirus E1A Proteins / Q000235:genetics; D017501:Adenovirus E1B Proteins / Q000235:genetics; D000260:Adenoviruses, Human / Q000235:genetics / Q000472:pathogenicity*; D000818:Animals; D018932:Chemokine CCL2 / Q000378:metabolism; D018925:Chemokines / Q000235:genetics; D003315:Cornea / Q000378:metabolism / Q000473:pathology / Q000821:virology; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D015828:Eye Infections, Viral / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D007634:Keratitis / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009195:Peroxidase / Q000378:metabolism; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017500:Adenovirus E1A Proteins; D017501:Adenovirus E1B Proteins; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D018925:Chemokines; D012333:RNA, Messenger; D012367:RNA, Viral; D009195:Peroxidase