Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines.
The use of DNA to deliver vaccine antigens offers many advantages, including ease of manufacture and cost. However, most DNA vaccines are plasmids and must be grown in bacterial culture, necessitating elements that are either unnecessary for effective gene delivery (for example, bacterial origins of replication) or undesirable (for example, antibiotic resistance genes). Removing these elements may improve the safety profile of DNA for the delivery of vaccines. Here, we describe a novel, double-stranded, linear DNA construct produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, polyA tail and telomeric ends. We compared these constructs (called 'Doggybones' because of their shape) with conventional plasmid DNA. Using luciferase-expressing constructs, we demonstrated that expression levels were equivalent between Doggybones and plasmids both in vitro and in vivo. When mice were immunized with DNA constructs expressing the HIV envelope protein gp140, equivalent humoral and cellular responses were induced. Immunizations with either construct type expressing hemagluttinin were protective against H1N1 influenza challenge. This is the first example of an effective DNA vaccine, which can be produced on a large scale by enzymatic processes.
Mucosal Infection & Immunity Group, Section of Infectious Diseases, Department of Medicine, Imperial College London, St Mary's Campus, London, UK.
A A
AA
Walters
Mucosal Infection & Immunity Group, Section of Infectious Diseases, Department of Medicine, Imperial College London, St Mary's Campus, London, UK.
E
E
Kinnear
Mucosal Infection & Immunity Group, Section of Infectious Diseases, Department of Medicine, Imperial College London, St Mary's Campus, London, UK.
R J
RJ
Shattock
Mucosal Infection & Immunity Group, Section of Infectious Diseases, Department of Medicine, Imperial College London, St Mary's Campus, London, UK.
J U
JU
McDonald
Touchlight Genetics Ltd., Leatherhead Food Research Institute, Leatherhead, Surrey, UK.
L J
LJ
Caproni
Touchlight Genetics Ltd., Leatherhead Food Research Institute, Leatherhead, Surrey, UK.
N
N
Porter
Mucosal Infection & Immunity Group, Section of Infectious Diseases, Department of Medicine, Imperial College London, St Mary's Campus, London, UK.
J S
0000000180938741
JS
Tregoning
D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D004247:DNA / Q000235:genetics* / Q000276:immunology; D004285:Dogs; D005260:Female; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000276:immunology; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology*; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology / Q000502:physiology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000276:immunology* / Q000821:virology; D010957:Plasmids / Q000235:genetics* / Q000276:immunology; D011061:Poly A / Q000235:genetics; D011401:Promoter Regions, Genetic; D016615:Telomere / Q000235:genetics; D014611:Vaccination; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000276:immunology*
D006389:Hemagglutinins, Viral; D019444:Vaccines, DNA; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1; D011061:Poly A; D004247:DNA