Vaccine efficacy of Salmonella strains expressing glycoprotein 63 with different promoters

Abstract

The development of Salmonella vaccine vectors has been hindered by both the requirement for multiple doses to induce immune responses and a lack of plasmid stability. Direct comparisons of different promoter systems with the same antigen are necessary to address these important issues. We have previously described an AroA- AroD- deletion mutant of Salmonella typhimurium (GID101) which expresses the gene encoding the Leishmania major promastigote surface glycoprotein gp63 (GID101). While this construct provided significant protection against L. major challenge to highly susceptible BALB/c mice, this required at least two oral doses. We report here the use of two different inducible promoters, the nirB and osmC promoters, to improve vaccine efficacy. These constructs (termed GID105 and GID106, respectively) expressed gp63 in vitro under inducible conditions and colonized BALB/c mice after oral administration. GID105 demonstrated greater plasmid stability in vitro and in vivo than did either GID106 or GID101, which expresses gp63 constitutively. Spleen and lymph node cells from mice immunized with a single oral dose of GID105 proliferated in vitro in response to L. major and secreted gamma interferon, whereas cells from mice given the other constructs did not. Mice immunized with a single oral dose of GID1O5 or GID106 developed significantly smaller lesions upon challenge with L. major, whereas mice administered GID101 did not. Mice administered GID105 also showed considerable resistance to Leishmania donovani infection. These data provide a direct comparison of promoter systems and demonstrate that the use of inducible promoters such as the nirB promoter allows a considerable improvement over the previous vaccine construct in terms of protection against infection.