A Recombinant Attenuated Yersinia pseudotuberculosis Vaccine Delivering a Y. pestis YopENt138-LcrV Fusion Elicits Broad Protection against Plague and Yersiniosis in Mice

Infect Immun. 2019 Sep 19;87(10):e00296-19. doi: 10.1128/IAI.00296-19. Print 2019 Oct.

Abstract

In this study, a novel recombinant attenuated Yersinia pseudotuberculosis PB1+ strain (χ10069) engineered with ΔyopK ΔyopJ Δasd triple mutations was used to deliver a Y. pestis fusion protein, YopE amino acid 1 to 138-LcrV (YopENt138-LcrV), to Swiss Webster mice as a protective antigen against infections by yersiniae. χ10069 bacteria harboring the pYA5199 plasmid constitutively synthesized the YopENt138-LcrV fusion protein and secreted it via the type 3 secretion system (T3SS) at 37°C under calcium-deprived conditions. The attenuated strain χ10069(pYA5199) was manifested by the establishment of controlled infection in different tissues without developing conspicuous signs of disease in histopathological analysis of microtome sections. A single-dose oral immunization of χ10069(pYA5199) induced strong serum antibody titers (log10 mean value, 4.2), secretory IgA in bronchoalveolar lavage (BAL) fluid from immunized mice, and Yersinia-specific CD4+ and CD8+ T cells producing high levels of tumor necrosis factor alpha (TNF-α), gamma interferon (IFN-γ), and interleukin 2 (IL-2), as well as IL-17, in both lungs and spleens of immunized mice, conferring comprehensive Th1- and Th2-mediated immune responses and protection against bubonic and pneumonic plague challenges, with 80% and 90% survival, respectively. Mice immunized with χ10069(pYA5199) also exhibited complete protection against lethal oral infections by Yersinia enterocolitica WA and Y. pseudotuberculosis PB1+. These findings indicated that χ10069(pYA5199) as an oral vaccine induces protective immunity to prevent bubonic and pneumonic plague, as well as yersiniosis, in mice and would be a promising oral vaccine candidate for protection against plague and yersiniosis for human and veterinary applications.

Keywords: Y. pseudotuberculosis; mucosal immunity; oral immunization; plague; systemic immunity; yersiniosis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Oral
  • Animals
  • Antibodies, Bacterial / biosynthesis*
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology
  • CD4-Positive T-Lymphocytes / drug effects
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / microbiology
  • CD8-Positive T-Lymphocytes / drug effects
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / microbiology
  • Cross Protection
  • Female
  • Gene Expression
  • Humans
  • Immunization
  • Immunoglobulin A / biosynthesis*
  • Interferon-gamma / genetics
  • Interferon-gamma / immunology
  • Interleukin-2 / genetics
  • Interleukin-2 / immunology
  • Lung / drug effects
  • Lung / immunology
  • Lung / microbiology
  • Male
  • Mice
  • Plague / immunology
  • Plague / microbiology
  • Plague / mortality
  • Plague / prevention & control*
  • Plague Vaccine / administration & dosage*
  • Plague Vaccine / biosynthesis
  • Plague Vaccine / genetics
  • Plague Vaccine / immunology
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Pore Forming Cytotoxic Proteins / genetics
  • Pore Forming Cytotoxic Proteins / immunology
  • Recombinant Fusion Proteins / administration & dosage*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Survival Analysis
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / immunology
  • Vaccines, Synthetic
  • Yersinia pestis / drug effects*
  • Yersinia pestis / immunology
  • Yersinia pestis / pathogenicity
  • Yersinia pseudotuberculosis / drug effects*
  • Yersinia pseudotuberculosis / immunology
  • Yersinia pseudotuberculosis / pathogenicity
  • Yersinia pseudotuberculosis Infections / immunology
  • Yersinia pseudotuberculosis Infections / microbiology
  • Yersinia pseudotuberculosis Infections / mortality
  • Yersinia pseudotuberculosis Infections / prevention & control*

Substances

  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Bacterial Outer Membrane Proteins
  • Immunoglobulin A
  • Interleukin-2
  • LcrV protein, Yersinia
  • Plague Vaccine
  • Pore Forming Cytotoxic Proteins
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • Vaccines, Synthetic
  • yopE protein, Yersinia
  • Interferon-gamma