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D. Zhu, X. Zhou, J. Chodosh; Characterization of Resident Corneal Leukocytes in Adenovirus Type 37 Infection. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3880.
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To characterize the changes in distribution, morphology, and maturation of resident antigen-presenting cells (APCs) of the corneal stroma in response to human adenovirus type 37 (HAdV-37) infection using a mouse model.
HAdV-37 or Cy3-labeled HAdV-37 was injected into the corneal stroma of C57BL/6J mice using a gas-powered microinjection system and glass micropipette needle. Control corneas were untouched or injected with dialysis buffer control. Mice were sacrificed at 30 min, 4 hr, 16 hr, and 24 hr post infection. Corneas were immunostained for CD11b (myeloid cell), CD11c (dendritic cell), CD86 (costimulatory molecule), and F4/80 (macrophage), and whole-mounted for confocal microscopy. Flow cytometry was also performed using anti-CD86 and CD11c antibodies at 16 hr post infection.
Untouched corneas revealed a large population of round and spindle-shaped CD11b+ and CD11c+ cells in the anterior stroma; a smaller CD11b+ population was seen in the posterior stroma. The density of CD11b+ and CD11c+ cells decreased from the periphery towards the center. Star-shaped CD86+ cells were seen only in the periphery, suggesting the localization of immature myeloid cells (CD11b+CD86-) in the central cornea. At 30 min post infection, Cy3-HAdV-37 was noted in CD11b+/-, CD11c+/-, and CD86+/-, but not F4/80+ cells. As early as 4 hr post-infection, upregulation of CD86 was observed in the corneal periphery indicating the possible maturation of resident APCs. At 16 hr post infection, CD86+ cells were observed in the paracentral cornea. Notably, a large influx of small, multi-lobed, CD11b+CD11c-CD86- cells (neutrophils) were seen throughout the stroma of infected corneas at 16 hr post infection. The observation of CD86 upregulation and neutrophil infiltration was validated by flow cytometry.
These data suggest that the maturation of resident corneal APCs may be a relatively early event in response to adenovirus infection in the cornea. Moreover, HAdV-37 appears capable of infecting all resident corneal cells except for F4/80+ macrophages.
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