Topical treatment with antisense oligonucleotides targeting tumor necrosis factor-alpha in herpetic stromal keratitis.
OBJECTIVE
Tumor necrosis factor (TNF)-alpha is a pleiotropic factor that is critical for the development of inflammation. The authors investigated whether topical application of TNF-alpha-antagonizing molecules, antisense oligonucleotides (ASON), might be an effective way of modifying the course of immune-mediated herpetic stromal keratitis (HSK).


METHODS
ASON targeting TNF-alpha mRNA were examined for their efficiency in interfering with the production of this cytokine in vitro. In vivo uptake was determined by FITC-labeled ASON. HSV-1 corneally infected mice were injected three times with ASON. Mice from the control groups received unrelated control oligonucleotides (CON) or buffer. The clinical course of HSK, the delayed-type hypersensitivity (DTH) reaction, the uptake of [(3)H]thymidine from cells derived from the spleen, virus-neutralizing antibody titers in the serum, and viral replication in the infected eyes were determined. The eyes were examined histologically. The corneal TNF-alpha content was measured by ELISA.


RESULTS
The TNF-alpha ASON reduced the lymphocytic cytokine expression in vitro. In vivo, the FITC-labeled molecules were detected in the cornea even after 10 days. In the TNF-alpha ASON mice the incidence of HSK decreased, and the severity of the disease was diminished. The corneal content of TNF-alpha was reduced, and the number of inflammatory cells was decreased. The other investigated parameters were not significantly altered by TNF-alpha ASON treatment.


CONCLUSIONS
The data suggest that TNF-alpha ASON diminishes the release of TNF-alpha from cultured lymphocytes and from lymphocytes in the HSV-1-infected cornea. This topical treatment mitigates the course of HSK, whereas the systemic antiviral effector functions were not impaired.
Wasmuth
Susanne
S
Department of Ophthalmology, University of Essen, Essen, Germany.
Bauer
Dirk
D
Yang
Yanning
Y
Steuhl
Klaus-Peter
KP
Heiligenhaus
Arnd
A
D000287:Administration, Topical; D000818:Animals; D003319:Corneal Stroma / Q000378:metabolism / Q000821:virology*; D016503:Drug Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D018259:Herpesvirus 1, Human / Q000502:physiology; D006968:Hypersensitivity, Delayed / Q000276:immunology; D016849:Keratitis, Herpetic / Q000188:drug therapy* / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha