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DNA vaccines have advantages over traditional vaccine modalities; however the relatively low immunogenicity restrains its translation into clinical use. Further optimizations are needed to get the immunogenicity of DNA vaccine closer to the level required for human use. Here we show that intramuscularly inoculating into a different limb each time significantly improves the immunogenicities of both DNA and recombinant vaccinia vaccines during multiple vaccinations, compared to repeated vaccination on the same limb. We term this strategy successive site translocating inoculation ( SSTI ).  SSTI  could work in synergy with genetic adjuvant and DNA prime-recombinant vaccinia boost regimen. By comparing  in vivo  antigen expression, we found that  SSTI  avoided the specific inhibition of  in vivo  antigen expression, which was observed in the limbs being repeatedly inoculated. Employing  in vivo  T cell depletion and passive IgG transfer, we delineated that the inhibition was not mediated by CD8 +  T cells but by specific antibodies. Finally, by using C3 −/−  mouse model and  in vivo  NK cells depletion, we identified that specific antibodies negatively regulated the  in vivo  antigen expression primarily in a complement depended way.

Successive site translocating inoculation potentiates DNA/recombinant vaccinia vaccination