title: Synthetic delivery system for tuberculosis vaccines: immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable PLG microparticles.
abstract: Tuberculosis remains a major public health burden which could be ameliorated by effective and well-defined subunit vaccines, particularly because the protective efficacy of current M. bovis BCG vaccines is both unpredictable and variable. The immunodominant 38 kDa antigen from Mycobacterium tuberculosis was entrapped in biodegradable poly (DL-lactide co-glycolide) (PLG) microparticles which served as a delivery system. Both cellular and humoral immune responses were assessed and compared with those obtained after immunization with the 38 kDa protein emulsified in incomplete Freund's adjuvant (IFA). Vaccination of mice with a single dose of antigen-loaded microparticles resulted in specific IgG titres peaking after five weeks comparable to those achieved after vaccination with protein emulsified in incomplete Freund's adjuvant (IFA). T-cell responses were found to be superior to those induced with antigen/IFA. The T- and B-cell epitope specificities ad judged with synthetic peptides were identical following immunization with antigen in microparticles or IFA. Differences in adjuvanticity were revealed by measuring antigen-specific IgG1, IgG2a and antigen-induced IFN-gamma secretion in vitro: substantially higher titres of IgG2a were observed following immunization with antigen/microparticles than with 38 kDa protein/IFA. This was paralleled by a tenfold higher secretion of IFN-gamma in mice injected with antigen/microparticles. Reduction in colony-forming units was not consistent in mice immunized with 38 kDa protein entrapped in microparticles which were subsequently infected with live tubercle bacilli. Taken together these results indicate that biodegradable PLG microparticles constitute a favorable candidate vaccine delivery system worthy of further assessment in the quest to develop better and defined agents protecting against tuberculosis.
authors.affiliation: MRC Clinical Sciences Centre, Hammersmith Hospital, London, UK.
authors.forename: H M
authors.initials: HM
authors.lastname: Vordermeier
authors.forename: A G
authors.initials: AG
authors.lastname: Coombes
authors.forename: P
authors.initials: P
authors.lastname: Jenkins
authors.forename: J P
authors.initials: JP
authors.lastname: McGee
authors.forename: D T
authors.initials: DT
authors.lastname: O'Hagan
authors.forename: S S
authors.initials: SS
authors.lastname: Davis
authors.forename: M
authors.initials: M
authors.lastname: Singh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial / Q000276:immunology*; D001500:BCG Vaccine / Q000008:administration & dosage* / Q000276:immunology; D002214:Capsules; D005069:Evaluation Studies as Topic; D005260:Female; D007132:Immunoglobulin Isotypes / Q000032:analysis / Q000378:metabolism; D019344:Lactic Acid; D008213:Lymphocyte Activation / Q000187:drug effects; D008222:Lymphokines / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010316:Particle Size; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000008:administration & dosage*; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014376:Tuberculosis / Q000517:prevention & control
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001500:BCG Vaccine; D002214:Capsules; D007132:Immunoglobulin Isotypes; D008222:Lymphokines; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid