Skin electroporation: effects on transgene expression, DNA persistence and local tissue environment.
BACKGROUND
Electrical pulses have been used to enhance uptake of molecules into living cells for decades. This technique, often referred to as electroporation, has become an increasingly popular method to enhance in vivo DNA delivery for both gene therapy applications as well as for delivery of vaccines against both infectious diseases and cancer. In vivo electrovaccination (gene delivery followed by electroporation) is currently being investigated in several clinical trials, including DNA delivery to healthy volunteers. However, the mode of action at molecular level is not yet fully understood.


RESULTS
This study investigates intradermal DNA electrovaccination in detail and describes the effects on expression of the vaccine antigen, plasmid persistence and the local tissue environment. Gene profiling of the vaccination site showed that the combination of DNA and electroporation induced a significant up-regulation of pro-inflammatory genes. In vivo imaging of luciferase activity after electrovaccination demonstrated a rapid onset (minutes) and a long duration (months) of transgene expression. However, when the more immunogenic prostate specific antigen (PSA) was co-administered, PSA-specific T cells were induced and concurrently the luciferase expression became undetectable. Electroporation did not affect the long-term persistence of the PSA-expressing plasmid.


CONCLUSIONS
This study provides important insights to how DNA delivery by intradermal electrovaccination affects the local immunological responses of the skin, transgene expression and clearance of the plasmid. As the described vaccination approach is currently being evaluated in clinical trials, the data provided will be of high significance.
Roos
Anna-Karin
AK
Department of Oncology and Pathology, Cancer Center Karolinska R8:01, Immune and Gene Therapy Laboratory, Karolinska Institute, Stockholm, Sweden. roos.anki@gmail.com
Eriksson
Fredrik
F
Timmons
James A
JA
Gerhardt
Josefine
J
Nyman
Ulrika
U
Gudmundsdotter
Lindvi
L
Bråve
Andreas
A
Wahren
Britta
B
Pisa
Pavel
P
D000818:Animals; D004247:DNA / Q000378:metabolism*; D004594:Electrophysiology / Q000379:methods; D018274:Electroporation; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D007271:Injections, Intradermal / Q000379:methods*; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000378:metabolism; D012867:Skin / Q000378:metabolism*; D019076:Transgenes
D004247:DNA