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The combination of TLR‐9 adjuvantation and electroporation‐mediated delivery enhances in vivo antitumor responses after vaccination with HPV‐16 E7 encoding DNA

Therapeutic DNA vaccination is an attractive adjuvant option to conventional methods in the fight against cancer, like surgery radiotherapy and chemotherapy. Despite strong antitumor effects that were observed in small animals with different antigens, DNA‐based vaccines remain weakly immunogenic in large animals and primates compared to protein‐based vaccines. Here, we sought to enhance the immunogenicity of a therapeutic nontransforming cervical cancer DNA vaccine (HPV‐16 E7SH) by introduction of a highly optimized CpG cassette into the plasmid backbone as well as by an optimized DNA delivery using an advanced electroporation (EP) technology. By integrating the means for agent administration and EP into a single device, this technology enables a simple, one‐step procedure that facilitates reproducibility. We found that highly optimized CpG motifs alone triggers an enhanced IFN‐γ and granzyme B response in Elispot assays as well as stronger tumor regression. Furthermore, these effects could be dramatically enhanced when the CpG cassette containing plasmid was administered via the newly developed EP technology. These data suggest that an optimized application of CpG‐enriched DNA vaccines may be an attractive strategy for the treatment of cancer. Collectively, these results provide a basis for the transfer of preclinical therapeutic DNA‐based immunization studies into successful clinical cancer trials.