Efficacy of intracellular activated promoters for generation of Salmonella-based vaccines

Infect Immun. 2010 Nov;78(11):4828-38. doi: 10.1128/IAI.00298-10. Epub 2010 Aug 23.

Abstract

Salmonella enterica is a versatile vaccine carrier for heterologous antigens. One strategy for vaccine antigen delivery is the use of live attenuated S. enterica strains that translocate heterologous antigens into antigen-presenting cells by means of type III secretion systems (T3SS). The feasibility of this approach has been demonstrated in various experimental vaccination studies. The efficacy of recombinant live vaccines is critically influenced by the optimal level of attenuation and many other factors. For the rational design of approaches involving translocation by T3SS, additional parameters are the level of expression of the heterologous antigens and the selection of carrier proteins for the delivery of antigens to desirable subcellular compartments of the target cell. We deployed the Salmonella pathogenicity island 2 (SPI2)-encoded T3SS for antigen delivery. The SPI2-T3SS and effector proteins are encoded by members of the large SsrAB regulon, including promoters with highly variable strength of expression. We investigated the effect of various in vivo-activated promoters of the SsrAB regulon on the efficacy of recombinant Salmonella vaccines. We observed that the use of promoters with higher strength results in greater synthesis of recombinant antigens and greater stimulation of T-cell responses in cell culture assays for the stimulation of T cells by the model antigen ovalbumin. In contrast, in vaccination experiments, promoters with a low level of expression resulted in the induction of higher amounts of T cells reactive to the model antigen listeriolysin. These results demonstrate that high-level expression of heterologous antigens does not necessarily result in optimal stimulation of immune responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens / genetics
  • Antigens / immunology
  • Antigens / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Bone Marrow Cells
  • Cells, Cultured
  • Dendritic Cells / microbiology*
  • Gene Expression Regulation*
  • Macrophages / microbiology*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Ovalbumin / genetics
  • Ovalbumin / immunology
  • Plasmids
  • Promoter Regions, Genetic*
  • RNA, Bacterial / genetics
  • RNA, Bacterial / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism*
  • Regulon
  • Salmonella typhimurium / genetics*
  • Salmonella typhimurium / metabolism
  • Salmonella typhimurium / pathogenicity
  • T-Lymphocytes / immunology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Vaccination
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology
  • Vaccines, Synthetic / metabolism*

Substances

  • Antigens
  • Bacterial Proteins
  • Membrane Proteins
  • RNA, Bacterial
  • Recombinant Proteins
  • SPI-2 protein, Salmonella
  • SsrB protein, Salmonella typhimurium
  • Transcription Factors
  • Vaccines, Synthetic
  • tmRNA
  • Ovalbumin