Methods Of Dna Vaccine Delivery 31 Electroporation

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Elecroporation is a common in vitro transfection method. Through the application of electric pulses, the cellular membrane is temporarily disrupted. This, combined with physical translocation of ionic plasmid DNA (ionophoresis), can result in efficient gene transfection. In vivo, the application of electroporation involves the use of probes or clamp electrodes to the site of plasmid administration. This method has been shown to induce long-term expression of reporter gene in vivo. Adapting the process parameters from low voltage, long pulses to high voltage, short pulses resulted in a 500x increase in expression in muscle (20).

Studies on the delivery of TAA antigens using electroporation have yielded promising results. Mendiratta et al. reported vaccination using electroporation with both plasmid encoded human GP100 and mouse TPR2 antigen elicited complete protection from melanoma challenge (21). Lohr et al. demonstrated that introduction of plasmid encoding IL-2 and IL-12 (inflammatory cytokine signals) by electroporation at tumor sites caused transduction and inhibition of murine melanoma without the systemic cytokine levels experienced after adenoviral gene transfer (22). Further investigations have shown that electroporation can be used to facilitate the discovery of novel antigen encoded plasmid constructs. For example, Kalat et al. used electroporation methods to optimize tyrosinase related protein-2 antigens to elicit CD8+ responses and inhibition of melanoma growth in two challenge models (23). This same group later demonstrated that electroporation was capable of inducing immune responses comparable to that of viral infection (24).

It has been suggested that increase in gene transfection is responsible for the amplification of immune responses observed, but it is also possible that tissue damage which occurs at the immunization site may recruit APCs, effectively increasing immuno-genicity (25). Unfortunately, electroporation can be destructive to tissues, and some have reported pain in patients during clinical trials (26).

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