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For reasons of efficiency  Escherichia coli  is used today as the microbial factory for production of plasmid DNA vaccines. To avoid hazardous antibiotic resistance genes and endotoxins from plasmid systems used nowadays, we have developed a system based on the food-grade  Lactococcus lactis  and a plasmid without antibiotic resistance genes. We compared the  L. lactis  system to a traditional one in  E. coli  using identical vaccine constructs encoding the gp120 of HIV-1. Transfection studies showed comparable gp120 expression levels using both vector systems. Intramuscular immunization of mice with  L. lactis  vectors developed comparable gp120 antibody titers as mice receiving  E. coli  vectors. In contrast, the induction of the cytolytic response was lower using the  L. lactis  vector. Inclusion of CpG motifs in the plasmids increased T-cell activation more when the  E. coli  rather than the  L. lactis  vector was used. This could be due to the different DNA content of the vector backbones. Interestingly, stimulation of splenocytes showed higher adjuvant effect of the  L. lactis  plasmid. The study suggests the developed  L. lactis  plasmid system as new alternative DNA vaccine system with improved safety features. The different immune inducing properties using similar gene expression units, but different vector backbones and production hosts give information of the adjuvant role of the silent plasmid backbone. The results also show that correlation between the  in vitro  adjuvanticity of plasmid DNA and its capacity to induce cellular and humoral immune responses in mice is not straight forward.

Immunological analysis of a Lactococcus lactis-based DNA vaccine expressing HIV gp120