title: Expression and immunogenicity of the mycobacterial Ag85B/ESAT-6 antigens produced in transgenic plants by elastin-like peptide fusion strategy.
abstract: This study explored a novel system combining plant-based production and the elastin-like peptide (ELP) fusion strategy to produce vaccinal antigens against tuberculosis. Transgenic tobacco plants expressing the mycobacterial antigens Ag85B and ESAT-6 fused to ELP (TBAg-ELP) were generated. Purified TBAg-ELP was obtained by the highly efficient, cost-effective, inverse transition cycling (ICT) method and tested in mice. Furthermore, safety and immunogenicity of the crude tobacco leaf extracts were assessed in piglets. Antibodies recognizing mycobacterial antigens were produced in mice and piglets. A T-cell immune response able to recognize the native mycobacterial antigens was detected in mice. These findings showed that the native Ag85B and ESAT-6 mycobacterial B- and T-cell epitopes were conserved in the plant-expressed TBAg-ELP. This study presents the first results of an efficient plant-expression system, relying on the elastin-like peptide fusion strategy, to produce a safe and immunogenic mycobacterial Ag85B-ESAT-6 fusion protein as a potential vaccine candidate against tuberculosis.
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466 Gatersleben, Germany.
authors.forename: Doreen Manuela
authors.initials: DM
authors.lastname: Floss
authors.forename: Michael
authors.initials: M
authors.lastname: Mockey
authors.forename: Galliano
authors.initials: G
authors.lastname: Zanello
authors.forename: Damien
authors.initials: D
authors.lastname: Brosson
authors.forename: Marie
authors.initials: M
authors.lastname: Diogon
authors.forename: Roger
authors.initials: R
authors.lastname: Frutos
authors.forename: Timothée
authors.initials: T
authors.lastname: Bruel
authors.forename: Valérie
authors.initials: V
authors.lastname: Rodrigues
authors.forename: Edwin
authors.initials: E
authors.lastname: Garzon
authors.forename: Claire
authors.initials: C
authors.lastname: Chevaleyre
authors.forename: Mustapha
authors.initials: M
authors.lastname: Berri
authors.forename: Henri
authors.initials: H
authors.lastname: Salmon
authors.forename: Udo
authors.initials: U
authors.lastname: Conrad
authors.forename: Laurence
authors.initials: L
authors.lastname: Dedieu
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002417:Cattle; D048708:Cell Growth Processes / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D004549:Elastin / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006968:Hypersensitivity, Delayed; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D010455:Peptides / Q000235:genetics; D018515:Plant Leaves / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D013154:Spleen / Q000166:cytology; D013552:Swine; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C093276:ESAT-6 protein, Mycobacterium tuberculosis; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; C065036:antigen 85, Mycobacterium bovis; D004549:Elastin