title: LcrV delivered via type III secretion system of live attenuated Yersinia pseudotuberculosis enhances immunogenicity against pneumonic plague.
abstract: Here, we constructed a Yersinia pseudotuberculosis mutant strain with arabinose-dependent regulated and delayed shutoff of crp expression (araC P(BAD) crp) and replacement of the msbB gene with the Escherichia coli msbB gene to attenuate it. Then, we inserted the asd mutation into this construction to form χ10057 [Δasd-206 ΔmsbB868::P(msbB) msbB(EC) ΔP(crp21)::TT araC P(BAD) crp] for use with a balanced-lethal Asd-positive (Asd(+)) plasmid to facilitate antigen synthesis. A hybrid protein composed of YopE (amino acids [aa]1 to 138) fused with full-length LcrV (YopE(Nt138)-LcrV) was synthesized in χ10057 harboring an Asd(+) plasmid (pYA5199, yopE(Nt138)-lcrV) and could be secreted through a type III secretion system (T3SS) in vitro and in vivo. Animal studies indicated that mice orally immunized with χ10057(pYA5199) developed titers of IgG response to whole-cell lysates of Y. pestis (YpL) and subunit LcrV similar to those seen with χ10057(pYA3332) (χ10057 plus an empty plasmid). However, only immunization of mice with χ10057(pYA5199) resulted in a significant secretory IgA response to LcrV. χ10057(pYA5199) induced a higher level of protection (80% survival) against intranasal (i.n.) challenge with ~240 median lethal doses (LD50) (2.4 × 10(4) CFU) of Y. pestis KIM6+(pCD1Ap) than χ10057(pYA3332) (40% survival). Splenocytes from mice vaccinated with χ10057(pYA5199) produced significant levels of gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin-17 (IL-17) after restimulation with LcrV and YpL antigens. Our results suggest that it is possible to use an attenuated Y. pseudotuberculosis strain delivering the LcrV antigen via the T3SS as a potential vaccine candidate against pneumonic plague.
authors.affiliation: Center for Infectious Disease and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, Arizona, USA wei.sun.1@asu.edu.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.affiliation: Center for Infectious Disease and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, Arizona, USA.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Sanapala
authors.affiliation: The Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas, USA.
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Henderson
authors.affiliation: Center for Infectious Disease and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, Arizona, USA.
authors.forename: Shandiin
authors.initials: S
authors.lastname: Sam
authors.affiliation: Center for Infectious Disease and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, Arizona, USA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Olinzock
authors.affiliation: The Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas, USA.
authors.forename: M Stephen
authors.initials: MS
authors.lastname: Trent
authors.affiliation: Center for Infectious Disease and Vaccinology, The Biodesign Institute, Arizona State University, Tempe, Arizona, USA School of Life Sciences, Arizona State University, Tempe, Arizona, USA.
authors.forename: Roy
authors.initials: R
authors.lastname: Curtiss
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D007071:Immunoglobulin A, Secretory / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000276:immunology; D051379:Mice; D010930:Plague / Q000276:immunology / Q000517:prevention & control*; D010931:Plague Vaccine / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics / Q000276:immunology*; D013154:Spleen / Q000276:immunology; D016019:Survival Analysis; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D016207:Cytokines; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; C043908:LcrV protein, Yersinia; D010931:Plague Vaccine; D052899:Pore Forming Cytotoxic Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic
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