title: RNA interference-mediated in vivo silencing of fas ligand as a strategy for the enhancement of DNA vaccine potency.
abstract: Intradermal administration of DNA vaccines encoding luciferase represents a convenient method to assess gene expression in vivo. Gene silencing by intradermal gene gun administration of DNA encoding short hairpin RNA (shRNA) may represent an effective technique for the specific knockdown of gene expression in vivo. In the current study, we characterized luciferase gene expression over time in vivo by noninvasive bioluminescence imaging. Furthermore, we characterized in vivo luciferase gene silencing with DNA encoding shRNA targeting luciferase. We also characterized human papillomavirus type 16 (HPV-16) E7-specific CD8(+) T cell immune responses in mice immunized with E7 DNA and DNA encoding shRNA targeting Fas ligand (FasL), a key proapoptotic signaling protein. Our results indicated that coadministration of DNA encoding shRNA targeting luciferase significantly reduced luciferase expression in mice intradermally administered luciferase DNA. Furthermore, we observed that mice vaccinated with E7-expressing DNA coadministered with DNA encoding shRNA targeting FasL generated significantly enhanced E7-specific CD8(+) cytotoxic T cell responses as well as potent therapeutic antitumor effects against E7-expressing tumors. Thus, intradermal administration of DNA encoding shRNA represents a plausible approach to silence genes in vivo and a potentially useful tool to enhance DNA vaccine potency.
authors.affiliation: Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD 21231, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Huang
authors.forename: Chih-Ping
authors.initials: CP
authors.lastname: Mao
authors.forename: Shiwen
authors.initials: S
authors.lastname: Peng
authors.forename: Chien-Fu
authors.initials: CF
authors.lastname: Hung
authors.forename: T-C
authors.initials: TC
authors.lastname: Wu
mesh_terms: D000818:Animals; D019470:Biolistics; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology; D053222:Fas Ligand Protein / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D052162:Human papillomavirus 16 / Q000276:immunology; D006801:Humans; D021621:Imaging, Three-Dimensional; D049409:Luciferases, Firefly / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000276:immunology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014162:Transfection; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*
chemical_list: D019496:Cancer Vaccines; D053222:Fas Ligand Protein; D050725:Papillomavirus E7 Proteins; D034741:RNA, Small Interfering; D019444:Vaccines, DNA; D049409:Luciferases, Firefly