title: In vivo expression of an interferon-alpha gene by intramuscular injection of naked DNA.
abstract: Acid-stable type I interferons belong to a multigene family. The biologic relevance of each subtype in vivo remains unknown. We have developed an experimental model in which muscles were transfected in situ with naked DNA plasmids encoding an IFN transgene to assess the roles of individual IFN subtypes in vivo. Murine IFN-alpha 9 gene was subcloned into several mammalian expression vectors. Adult C57BL/6 mice were injected bilaterally in regenerating tibialis anterior muscles with naked DNA 5 days after muscle injury to enhance DNA uptake and expression. In the muscles of mice given the IFN-alpha 9 plasmid constructs, acid-stable IFNs were detected by bioassay using reduction in cytopathic effect of encephalomyocarditis virus-infected L929 cells. In these same muscles, IFN-alpha 9 transcripts were identified by RT-PCR, indicating that transcription had occurred. Acid-stable IFNs were detected from days 7 to 28 post-DNA inoculation. Furthermore, these proteins were found in the sera of DNA-inoculated mice. Control groups of mice given the blank expression vectors did not produce detectable IFNs in muscle or sera as determined by bioassay, nor were transcripts detected by RT-PCR. This approach now allows investigation of the effector function of individual subtypes in various murine disease models.
authors.affiliation: Department of Microbiology, University of Western Australia, Queen Elizabeth II Medical Centre, Nedlands, Perth, Western Australia.
authors.forename: C M
authors.initials: CM
authors.lastname: Lawson
authors.forename: W S
authors.initials: WS
authors.lastname: Yeow
authors.forename: C M
authors.initials: CM
authors.lastname: Lee
authors.forename: M W
authors.initials: MW
authors.lastname: Beilharz
mesh_terms: D000818:Animals; D005260:Female; D007273:Injections, Intramuscular; D016898:Interferon-alpha / Q000097:blood / Q000145:classification / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D009132:Muscles / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection
chemical_list: D016898:Interferon-alpha; D012333:RNA, Messenger