A single dose of oral DNA immunization delivered by attenuated Salmonella typhimurium down-regulates transgene expression in HBsAg transgenic mice

Abstract

The efficacy of immunization with Salmonella typhimurium aroA to deliver the plasmid pRc/CMV-HBsAg (i.e. an oral DNA vaccine) was compared with that of intramuscular immunization with the same plasmid DNA, and with recombinant HBsAg protein, in a HBsAg transgenic mouse model. A single dose of oral DNA vaccine evoked vigorous Th1 cell and CTL responses and production of IgG2 subclass of anti-HBs after 2 weeks, and this was accompanied by a transient hepatitic flare with elevated alanine aminotransferase in the first 3 weeks. Concomitantly, the level of HBsAg-mRNA in liver tissues decreased by more than fourfold and viral-antigen expression was curtailed markedly in hepatocytes compared with controls. Hepatitic flare subsided after 3 weeks, but suppression of the transgene expression was continued in the absence of overt liver pathology for the remaining duration of the experiment (i.e. 12 weeks), and possibly beyond. The other vaccines could also break immune tolerance, but this was achieved only after repeated booster doses of the respective vaccines, and they did not affect transgene expression, or induce hepatic flare. We previously showed in non-transgenic mice that immunization by the oral DNA vaccine is achieved by an active intestinal infection with a bacterial carrier that is an adept intracellular parasite, and the immune response to the vaccination is orchestrated by phagocytic APC. Our present findings further implicated that the combined effects of an innate and a specific immune response induced by oral DNA vaccination are crucial in down-regulating HBsAg-transgene expression in hepatocytes.