title: Pulmonary immunization using antigen 85-B polymeric microparticles to boost tuberculosis immunity.
abstract: This study aims to evaluate immunization with polymeric microparticles containing recombinant antigen 85B (rAg85B) delivered directly to the lungs to protect against tuberculosis. rAg85B was expressed in Escherichia coli and encapsulated in poly(lactic-co-glycolic acid) microparticles (P-rAg85B). These were delivered as dry powders to the lungs of guinea pigs in single or multiple doses of homologous and heterologous antigens. Bacille Calmette-Guérin (BCG) delivered subcutaneously was employed as the positive control and as part of immunization strategies. Immunized animals were challenged with a low-dose aerosol of Mycobacterium tuberculosis (MTB) H37Rv to assess the extent of protection measured as reduction in bacterial burden (CFU) in the lungs and spleens of guinea pigs. Histopathological examination and morphometric analysis of these tissues were also performed. The heterologous strategy of BCG prime-P-rAg85B aerosol boosts appeared to enhance protection from bacterial infection, as indicated by a reduction in CFU in both the lungs and spleens compared with untreated controls. Although the CFU data were not statistically different from the BCG and BCG-BCG groups, the histopathological and morphometric analyses indicated the positive effect of BCG-P-rAg85B in terms of differences in area of tissue affected and number and size of granulomas observed in tissues. P-rAg85B microparticles appeared to be effective in boosting a primary BCG immunization against MTB infection, as indicated by histopathology and morphometric analysis. These encouraging observations are relevant to boosting adults previously immunized with BCG or exposed to MTB, commonly the case in the developing world, and should be followed by further assessment of an appropriate immunization protocol for maximum protection.
authors.affiliation: Molecular Pharmaceutics, School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7571, USA.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Lu
authors.forename: Lucila
authors.initials: L
authors.lastname: Garcia-Contreras
authors.forename: Pavan
authors.initials: P
authors.lastname: Muttil
authors.forename: Danielle
authors.initials: D
authors.lastname: Padilla
authors.forename: Ding
authors.initials: D
authors.lastname: Xu
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.forename: Miriam
authors.initials: M
authors.lastname: Braunstein
authors.forename: David N
authors.initials: DN
authors.lastname: McMurray
authors.forename: Anthony James
authors.initials: AJ
authors.lastname: Hickey
mesh_terms: D000336:Aerosols; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage*; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed; D019344:Lactic Acid / Q000008:administration & dosage*; D008168:Lung; D008863:Microspheres; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D011100:Polyglycolic Acid / Q000008:administration & dosage*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011994:Recombinant Proteins / Q000008:administration & dosage; D014376:Tuberculosis / Q000276:immunology*
chemical_list: D000336:Aerosols; D000942:Antigens, Bacterial; D011994:Recombinant Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid