title: Ultrastructure of adenovirus keratitis.
abstract: OBJECTIVE
  We determined the ultrastructure of mouse adenovirus keratitis, a model for human adenovirus keratitis.
  
  
  METHODS
  Adenovirus keratitis was induced in C57Bl/6j mice by intrastromal injection of human adenovirus species D type 37 (HAdV-D37) with a heat-pulled, glass, micropipette needle under compressed air. At select time points after infection, mice were euthanized and their corneas removed, fixed, and sectioned at 70-nm thickness for electron microscopy.
  
  
  RESULTS
  Injection of HAdV-D37 into the mouse corneal stroma placed virus predominantly in the pericellular corneal stromal matrix. Virus was seen bound to and entering stromal cells at 1 and 2 hours after infection, respectively. Cell membrane transit by virus was seen to involve two distinct structures resembling caveolae and macropinosomes. However, later during infection intracellular virus was not seen within membrane-bound organelles. By 8 hours after infection, intracellular virus had accumulated into densely packed, perinuclear arrays. Virus disassembly was not obvious at any time point after infection. Infiltrating neutrophils seen by one day after infection had engulfed degraded stromal cells by 4 days after infection.
  
  
  CONCLUSIONS
  By transmission electron microscopy, injected HAdV-D37 readily enters stromal cells in the C57Bl/6j mouse cornea and induces stromal inflammation, as was shown previously by light microscopy. However, electron microscopy also revealed dense, static arrays of intracytoplasmic virus, suggesting a block in viral capsid disassembly and viral DNA nuclear entry. These findings may explain why human adenoviruses do not replicate in the mouse corneal stroma.
authors.affiliation: Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Howe Laboratory, Harvard Medical School, Boston, Massachusetts, United States.
authors.forename: Santanu
authors.initials: S
authors.lastname: Mukherjee
authors.affiliation: Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Howe Laboratory, Harvard Medical School, Boston, Massachusetts, United States.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Howe Laboratory, Harvard Medical School, Boston, Massachusetts, United States.
authors.forename: Jaya
authors.initials: J
authors.lastname: Rajaiya
authors.affiliation: Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Howe Laboratory, Harvard Medical School, Boston, Massachusetts, United States.
authors.forename: James
authors.initials: J
authors.lastname: Chodosh
mesh_terms: D000256:Adenoviridae / Q000302:isolation & purification*; D000258:Adenovirus Infections, Human / Q000473:pathology* / Q000821:virology; D000818:Animals; D003319:Corneal Stroma / Q000648:ultrastructure* / Q000821:virology; D004195:Disease Models, Animal; D015828:Eye Infections, Viral / Q000473:pathology* / Q000821:virology; D005260:Female; D007634:Keratitis / Q000473:pathology* / Q000821:virology; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission / Q000379:methods
keywords: adenovirus; electron microscopy; keratitis; mouse