title: Recombinant Bacillus subtilis spores expressing MPT64 evaluated as a vaccine against tuberculosis in the murine model.
abstract: Recombinant Bacillus subtilis spores expressing a TB antigen, MPT64, were tested for their ability to protect mice against tuberculosis challenge. A chimeric gene consisting of the spore coat gene cotB fused to mpt64 was constructed, and expression of a stable CotB-MPT64 hybrid protein of the spore coat verified. Spores were evaluated as a live vaccine and also formaldehyde inactivated. Mice were given three doses of spores or alternatively used in a prime-boost regimen with BCG. The results showed that inactivated recombinant spores were able to reduce the bacterial burden in the lungs of mice to comparable levels to that of BCG. In the prime-boost regimen, both live and inactivated spores showed a reduction in bacterial load in comparison with BCG. ELISPOT and polyfunctional T-cell analysis were performed to examine cellular responses and showed that antigen-specific secretion of Th1 cytokines was stimulated after immunisation with inactive recombinant spores and BCG. In summary, recombinant spores can elicit Th1 responses, which are important for protection against TB disease.
authors.affiliation: School of Biological Sciences, Royal Holloway, University of London, Egham, UK.
authors.forename: Laura
authors.initials: L
authors.lastname: Sibley
authors.forename: Rajko
authors.initials: R
authors.lastname: Reljic
authors.forename: David S
authors.initials: DS
authors.lastname: Radford
authors.forename: Jen-Min
authors.initials: JM
authors.lastname: Huang
authors.forename: Huynh A
authors.initials: HA
authors.lastname: Hong
authors.forename: Rocky M
authors.initials: RM
authors.lastname: Cranenburgh
authors.forename: Simon M
authors.initials: SM
authors.lastname: Cutting
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001412:Bacillus subtilis / Q000235:genetics*; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000235:genetics; D061505:Cell Surface Display Techniques; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004337:Drug Carriers; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D005822:Genetic Vectors; D008168:Lung / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013171:Spores, Bacterial / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology; D014376:Tuberculosis / Q000276:immunology / Q000517:prevention & control*; D032581:Tuberculosis Vaccines / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D014611:Vaccination / Q000379:methods; D015164:Vaccines, Inactivated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C439051:CotB protein, Bacillus subtilis; D016207:Cytokines; D004337:Drug Carriers; C510537:MPT64 protein, Mycobacterium tuberculosis; D011993:Recombinant Fusion Proteins; D032581:Tuberculosis Vaccines; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic
keywords: TB; mucosal vaccine; tuberculosis; vaccine