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* Vaccine Research Group, Division of Microbiology, Gesellschaft fuer Biotechnologische Forschung-German Research Centre for Biotechnology, Braunschweig, Germany;
Institute of Microbiology and Genetics, Section Microbiology and Biotechnology, University of Vienna, Wien, Austria;
Biotech Innovation Research Development and Consulting (BIRD-C) GmbH and CoKEG, Vienna, Austria; and
Cancer Research Institute, Laboratory of Tumor Cell Biology, Bratislava, Slovakia
Mass implementation of DNA vaccines is hindered by the requirement of high plasmid dosages and poor immunogenicity. We evaluated the capacity of Mannheimia haemolytica ghosts as delivery system for DNA vaccines. In vitro studies showed that bacterial ghosts loaded with a plasmid carrying the green fluorescent protein-encoding gene (pEGFP-N1) are efficiently taken up by APC, thereby leading to high transfection rates (52–60%). Vaccination studies demonstrated that ghost-mediated delivery by intradermal or i.m. route of a eukaryotic expression plasmid containing the gene coding for 
-galactosidase under the control of the CMV immediate early gene promoter (pCMV) stimulates more efficient Ag-specific humoral and cellular (CD4+ and CD8+) immune responses than naked DNA in BALB/c mice. The use of ghosts also allows modulating the major Th response from a mixed Th1/Th2 to a more dominant Th2 pattern. Intravenous immunization with dendritic cells loaded ex vivo with pCMV-containing ghosts also resulted in the elicitation of -galactosidase-specific responses. This suggests that dendritic cells play an important role in the stimulation of immune responses when bacterial ghosts are used as a DNA delivery system. Bacterial ghosts not only target the DNA vaccine construct to APC, but also provide a strong danger signal, acting as natural adjuvants, thereby promoting efficient maturation and activation of dendritic cells. Thus, bacterial ghosts constitute a promising technology platform for the development of more efficient DNA vaccines.
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