A plasmid construct encoding murine interferon beta antagonizes the replication of herpes simplex virus type I in vitro and in vivo.
In the present study, we employed a plasmid DNA encoding murine interferon (IFN)-beta to assess its antiviral efficacy in an in vitro transfection-infection assay and in an ocular HSV-1 infection model of mice. In the in vitro assay, transfection of mouse fibroblasts with the IFN-beta transgene resulted in a 17-fold or greater reduction in the viral load of HSV-1 at a multiplicity of infection (MOI) of 1 compared to that of those mice treated with the plasmid control. RT-PCR analysis of representative immediate early (ICP27), early (thymidine kinase, TK) and late (VP16) viral genes found no changes in the level of expression comparing the IFN-beta transgene- to the vector-treated control group, suggesting that the IFN-beta transgene may act at the post-transcriptional level of viral replication. In the ocular HSV-1 infection model, topical application of the plasmid DNA encoding murine IFN-beta onto mouse cornea enhanced cumulative survival and significantly reduced the viral load of HSV-1 in the eyes and trigeminal ganglia of mice at both day 3 and 6 post-infection compared with mice treated with the plasmid vector control or normal saline. Neutralizing antibody to IFN-beta blocked the protective effect elicited by the IFN-beta transgene. Unlike the in vitro experiment, viral gene expression was reduced in the trigeminal ganglion of mice pre-treated 24 h with the IFN-beta transgene day 3 (ICP27 and VP16) and day 6 (ICP27, TK, DNA polymerase, and VP16) post-infection in comparison to mice treated with the plasmid vector control as determined by semi-quantitative RT-PCR.
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Department of Microbiology, Immunology, and Parasitology, Louisiana State University Health Sciences Center, New Orleans, LA, USA.
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D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage / Q000276:immunology; D002460:Cell Line; D003315:Cornea / Q000821:virology; D005260:Female; D005347:Fibroblasts; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics; D006561:Herpes Simplex / Q000235:genetics / Q000276:immunology / Q000628:therapy* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000276:immunology / Q000502:physiology*; D016899:Interferon-beta / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology / Q000502:physiology*; D051379:Mice; D008813:Mice, Inbred ICR; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics*; D015996:Survival Rate; D014162:Transfection; D019076:Transgenes / Q000235:genetics; D012668:Trigeminal Ganglion / Q000821:virology; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D019562:Viral Load; D014779:Virus Replication
D000998:Antiviral Agents; D019444:Vaccines, DNA; D016899:Interferon-beta