title: Early events of electroporation-mediated intramuscular DNA vaccination potentiate Th1-directed immune responses.
abstract: BACKGROUND
  Application of electrical pulses after DNA injection into muscle increases expression of the encoded genes, and is shown to improve antigen-specific immune responses when used for DNA vaccination. In addition, electroporation causes tissue injury and inflammatory reactions. Together with immune stimulatory motifs in the injected DNA these factors may potentiate the immune response by acting as adjuvants for the antigen. Here, we have examined the role of these factors in promoting the efficiency of DNA vaccination.
  
  
  METHODS
  We injected a plasmid DNA vector containing the gene Ag85B from M. tuberculosis into mouse quadriceps muscles followed by electroporation. Ag85B was under control of a Tet-responsive promoter, and was expressed either immediately or up to 28 days later by administrating doxycycline to the mice. Delayed expression was combined with injection of non-coding DNA or saline with or without electroporation to examine the ability of these factors to enhance the Ag85B-specific antibody response in the blood and cellular responses in the spleen. Blood samples were analysed with ELISA, while the number of Ag85B-specific IFN-gamma- and IL-4-producing spleenocytes was analysed with ELISpot.
  
  
  RESULTS
  Delaying Ag85B expression by 5 or 28 days caused lower anti-Ag85B-specific IgG2a levels. In contrast, the IgG1 antibody response was not significantly affected. Injection of non-coding DNA followed by electroporation moderately increased the IgG2a response. Delaying the Ag85B expression by 28 days reduced the average number of Ag85B-specific IFN-gamma-producing spleenocytes by over 60%. No significant change in the number of IL-4-producing Ag85B-specific spleenocytes was observed.
  
  
  CONCLUSIONS
  These results suggest that DNA and electroporation per se may act as good adjuvants in promoting efficient Th1-directed responses during DNA vaccination.
authors.affiliation: Department of Physiology, University of Oslo, Box 1103 Blindern, 0316 Oslo, Norway. eirik.gronevik@gmail.com
authors.forename: Eirik
authors.initials: E
authors.lastname: Grønevik
authors.forename: Iacob
authors.initials: I
authors.lastname: Mathiesen
authors.forename: Terje
authors.initials: T
authors.lastname: Lømo
mesh_terms: D000818:Animals; D002417:Cattle; D018274:Electroporation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D007074:Immunoglobulin G / Q000378:metabolism; D007273:Injections, Intramuscular / Q000379:methods; D007371:Interferon-gamma / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D013154:Spleen / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology*; D014162:Transfection; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D007074:Immunoglobulin G; D019444:Vaccines, DNA; D015847:Interleukin-4; D007371:Interferon-gamma; D008156:Luciferases