title: Oral immunogenicity of the plant proteinase bromelain.
abstract: Bromelain is a natural mixture of proteolytic enzymes derived from pineapple stem that has been shown to have anti-inflammatory activity when administered orally. Although most proteins given orally without adjuvant (e.g., food) result in tolerance, we previously reported that long-term oral exposure to bromelain stimulated the development of high serum anti-bromelain antibody titers. The purpose of these studies was to further investigate the mechanisms responsible for the immunogenicity of oral bromelain. Results showed that repeated exposure was required for development of anti-bromelain antibodies, with strong antibody responses in all mice that received at least 12 doses of bromelain either orally or intragastrically over 3-6 weeks. Proteolytic activity was required for strong oral immunogenicity in the absence of conventional adjuvant, with strong serum antibody responses generated against proteolytically active bromelain and trypsin, but not against ovalbumin, lysozyme, or inactivated bromelain. Significantly higher anti-bromelain antibody titers were seen in IL-10-deficient versus wild-type mice, suggesting that simultaneous treatments that decrease IL-10 activity may further enhance systemic antibody responses following oral exposure. The antibodies generated did not affect the proteolytic activity of bromelain. The data demonstrate that proteolytically active antigens such as bromelain can stimulate both systemic and mucosal immune responses following repeated oral exposure. Further studies of the mechanisms involved in generation of immune responses following oral exposure to proteolytically active antigens can lead to a better understanding of mechanisms of oral tolerance and to the development of novel adjuvants for oral vaccines.
authors.affiliation: Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA. laura.hale@duke.edu <laura.hale@duke.edu>
authors.forename: Laura P
authors.initials: LP
authors.lastname: Hale
authors.forename: David J
authors.initials: DJ
authors.lastname: Fitzhugh
authors.forename: Herman F
authors.initials: HF
authors.lastname: Staats
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000284:Administration, Oral; D031230:Ananas / Q000201:enzymology*; D000818:Animals; D001963:Bromelains / Q000008:administration & dosage / Q000276:immunology*; D002772:Cholera Toxin / Q000008:administration & dosage / Q000276:immunology; D005243:Feces / Q000737:chemistry; D005260:Female; D018928:Immunity, Mucosal / Q000276:immunology; D007070:Immunoglobulin A / Q000032:analysis / Q000737:chemistry; D007074:Immunoglobulin G / Q000097:blood; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009113:Muramidase / Q000276:immunology; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology; D010447:Peptide Hydrolases / Q000008:administration & dosage / Q000276:immunology; D018547:Plant Stems / Q000201:enzymology; D012463:Saliva / Q000737:chemistry; D014357:Trypsin / Q000008:administration & dosage / Q000276:immunology; D014611:Vaccination / Q000379:methods
chemical_list: D000276:Adjuvants, Immunologic; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D016753:Interleukin-10; D001963:Bromelains; D010047:Ovalbumin; D002772:Cholera Toxin; D009113:Muramidase; D010447:Peptide Hydrolases; D014357:Trypsin