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J. Lee, E. Jun, J. Sunwoo, H. Lee, E. Kim, J.-H. Kim, J. Kim, M. Kim, Y. Kim, H. Tchah; Ex-vivo Model of Coxsackievirus Conjunctivitis Using Multilayered Human Conjunctival Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3882.
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To establish the ex-vivo model of coxsackievirus conjunctivitis.
Human conjunctival epithelial cells were cultured for two weeks in serum free air-liquid interface system. Expression of CK 19 and MUC5AC in the cultured cells were studied by immunohistochemistry. Coxsackievirus A24 (CA24) was challenged for two hours in the cultured epithelial cells. Virus replication was measured in the cultured cells and media after 24 and 72 hours by TCID50 method. Histological changes were studied by H&E, DAPI and TUNEL staining.
Cultured human conjunctival epithelial cells formed multiple layers. Cultured cells showed positive immunohistochemical staining for CK19 and MUC5AC. Viral replication measured by TCID50 showed 10-fold increase at 72 hours compared to 24 hours. Mild cytopathic effect was found at 24 hours after viral inoculation. Cell damages with apoptotic features were remarkable at 72 hours.
Multilayered cells showed characteristics of conjunctival epithelial cells. Virus replicated in the cultured cells inducing cytopathic effects. This conjunctivitis model would be useful in the future study to investigate pathogenesis of acute hemorrhagic conjunctivitis and the effectiveness of antiviral treatment.
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