Programm                 "Degeneration und Regeneration– Grundlagen, Diagnostik und Therapie"

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Local Suppression of Tumor Necrosis Factor Alpha Improves Experimental Herpetic Keratitis

Wasmuth S.1,2, Bauer D.1, Yang Y.1, Steuhl K. P.2, Heiligenhaus A.1
1Ophtha-Lab and Department of Ophthalmology at St. Franziskus Hospital, Muenster; 2Department of Ophthalmology, University of Duisburg-Essen

Purpose: In this study we investigated if a local treatment with antisense oligonucleotides (ASON) targeting TNF alpha is useful to improve the course of herpes simplex keratitis (HSK).
Method: Cell suspensions prepared from spleens or regional lymph nodes from herpes simplex virus-1 infected mice were incubated with 0.5 to 20 µM of ASON, unrelated control oligonucleotides (CON) or medium. 24 h later, the cell culture supernatants were examined by ELISA for their content of TNF alpha. FITC-ASON were used to study the uptake of the oligonucleotides into the cells by FACScan analysis. In vivo, FITC-ASON or unbound FITC were injected into the corneas of BALB/c mice. After different periods of time, paraffin-sections of these eyes were examined by fluorescence microscopy. On day 0 mice were infected corneally with 100000 PFU HSV-1. On days -1, 1 and 4 the animals were treated topically with ASON targeting TNF-alpha. Mice from the control groups received CON or buffer. All animals were examined daily.
Results: The in vitro production of TNF alpha by cells from spleens and regional lymph nodes was strongly reduced following ASON treatment. In vivo, after a single injection of

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