title: Live attenuated Salmonella vaccines displaying regulated delayed lysis and delayed antigen synthesis to confer protection against Mycobacterium tuberculosis.
abstract: Live recombinant attenuated Salmonella vaccine (RASV) strains have great potential to induce protective immunity against Mycobacterium tuberculosis by delivering M. tuberculosis antigens. Recently, we reported that, in orally immunized mice, RASV strains delivering the M. tuberculosis early secreted antigenic target 6-kDa (ESAT-6) protein and culture filtrate protein 10 (CFP-10) antigens via the Salmonella type III secretion system (SopE amino-terminal region residues 1 to 80 with two copies of ESAT-6 and one copy of CFP-10 [SopE(Nt80)-E2C]) afforded protection against aerosol challenge with M. tuberculosis. Here, we constructed and evaluated an improved Salmonella vaccine against M. tuberculosis. We constructed translational fusions for the synthesis of two copies of ESAT-6 plus CFP-10 fused to the OmpC signal sequence (OmpC(SS)-E2C) and amino acids 44 to 338 of antigen 85A (Ag85A(294)) flanked by the signal sequence (SS) and C-terminal peptide (CT) of β-lactamase (Bla(SS)-Ag85A(294)-Bla(CT)) to enable delivery via the Salmonella type II secretion system. The genes expressing these proteins were cloned as an operon transcribed from P(trc) into isogenic Asd(+)/MurA(+) pYA3681 lysis vector derivatives with different replication origins (pBR, p15A, pSC101), resulting in pYA4890, pYA4891, and pYA4892 for SopE(Nt80)-E2C/Ag85A(294) synthesis and pYA4893 and pYA4894 for OmpC(SS)-E2C/Ag85A(294) synthesis. Mice orally immunized with the RASV χ11021 strain engineered to display regulated delayed lysis and regulated delayed antigen synthesis in vivo and harboring pYA4891, pYA4893, or pYA4894 elicited significantly greater humoral and cellular immune responses, and the RASV χ11021 strain afforded a greater degree of protection against M. tuberculosis aerosol challenge in mice than RASVs harboring any other Asd(+)/MurA(+) lysis plasmid and immunization with M. bovis BCG, demonstrating that RASV strains displaying regulated delayed lysis with delayed antigen synthesis resulted in highly immunogenic delivery vectors for oral vaccination against M. tuberculosis infection.
authors.affiliation: Center for Infectious Diseases and Vaccinology at Biodesign Institute, Arizona State University, Tempe, Arizona, USA.
authors.forename: María Dolores
authors.initials: MD
authors.lastname: Juárez-Rodríguez
authors.forename: Jiseon
authors.initials: J
authors.lastname: Yang
authors.forename: Rebin
authors.initials: R
authors.lastname: Kader
authors.forename: Praveen
authors.initials: P
authors.lastname: Alamuri
authors.forename: Roy
authors.initials: R
authors.lastname: Curtiss
authors.forename: Josephine E
authors.initials: JE
authors.lastname: Clark-Curtiss
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005260:Female; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D008168:Lung / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D022562:Salmonella Vaccines / Q000276:immunology*; D014376:Tuberculosis / Q000517:prevention & control; D032581:Tuberculosis Vaccines / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D022562:Salmonella Vaccines; D032581:Tuberculosis Vaccines