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Visceral leishmaniasis is a major neglected tropical disease, with an estimated 500,000 new cases and more than 50,000 deaths attributable to this disease every year. Drug therapy is available but costly and resistance against several drug classes has evolved. Despite all efforts, no commercial, let alone affordable, vaccine is available to date. Thus, the development of cost effective, needle-independent vaccines is a high priority. Here, we have continued efforts to develop live vaccine carriers based on recombinant  Salmonella . We used an  in silico  approach to select novel  Leishmania  parasite antigens from proteomic data sets, with selection criteria based on protein abundance, conservation across  Leishmania  species and low homology to host species. Five chosen antigens were differentially expressed on the surface or in the cytosol of  Salmonella typhimurium  SL3261. A two-step procedure was developed to select optimal  Salmonella  vaccine strains for each antigen, based on bacterial fitness and antigen expression levels. We show that vaccine strains of  Salmonella  expressing the novel  Leishmania  antigens LinJ08.1190 and LinJ23.0410 significantly reduced visceralisation of  L. major  and enhanced systemic resistance against  L. donovani  in susceptible BALB/c mice. The results show that  Salmonella  are valid vaccine carriers for inducing resistance against visceral leishmaniasis but that their use may not be suitable for all antigens.

Single Dose Novel Salmonella Vaccine Enhances Resistance against Visceralizing L. major and L. donovani Infection in Susceptible BALB/c Mice