title: Pulmonary immunization of guinea pigs with diphtheria CRM-197 antigen as nanoparticle aggregate dry powders enhance local and systemic immune responses.
abstract: This study establishes the immune response elicited in guinea pigs after pulmonary and parenteral immunizations with diphtheria CRM-197 antigen (CrmAg). Several spray-dried powders of formalin-treated/untreated CrmAg nanoaggregates with L-leucine were delivered to the lungs of guinea pigs. A control group consisting of alum with adsorbed CrmAg in saline was administered by intramuscular injection. Animals received three doses of powder vaccines containing 20 or 40 μg of CrmAg. The serum IgG titers were measured for 16 weeks after the initial immunization; IgA titers were measured at the time of sacrifice in the broncho-alveolar lavage fluid. Further, toxin neutralization tests in naïve guinea pigs were performed for a few select serum samples. Histopathology of the lung tissues was conducted to evaluate inflammation or injury to the lung tissues. While the highest titer of serum IgG antibody was observed in guinea pigs immunized by the intramuscular route, those animals immunized with dry powder formulation by the pulmonary route, and without the adjuvant alum, exhibited high IgA titers. A pulmonary administered dry powder, compared to parenteral immunization, conferred complete protection in the toxin neutralization test. Mild inflammation was observed in lung tissues of animals receiving dry powder vaccines by the pulmonary route. Thus, administering novel CrmAg as dry powders to the lungs may be able to overcome some of the disadvantages observed with the existing diphtheria vaccine which is administered by the parenteral route. In addition, these powders will have the advantage of eliciting a high mucosal immune response in the lungs without using traditional adjuvants.
authors.affiliation: University of North Carolina, Chapel Hill, 27599-7571, USA.
authors.forename: Pavan
authors.initials: P
authors.lastname: Muttil
authors.forename: Brian
authors.initials: B
authors.lastname: Pulliam
authors.forename: Lucila
authors.initials: L
authors.lastname: Garcia-Contreras
authors.forename: John Kevin
authors.initials: JK
authors.lastname: Fallon
authors.forename: Chenchen
authors.initials: C
authors.lastname: Wang
authors.forename: Anthony James
authors.initials: AJ
authors.lastname: Hickey
authors.forename: David A
authors.initials: DA
authors.lastname: Edwards
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology; D001992:Bronchoalveolar Lavage Fluid; D004167:Diphtheria Toxin / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D007111:Immunity, Cellular; D019344:Lactic Acid; D008168:Lung; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles; D009500:Neutralization Tests; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011208:Powders
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D004167:Diphtheria Toxin; D011208:Powders; C028581:CRM197 (non-toxic variant of diphtheria toxin); D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid