title: Generation of hypoallergenic DNA vaccines by forced ubiquitination: preventive and therapeutic effects in a mouse model of allergy.
abstract: BACKGROUND
  Hypoallergenic immunotherapy of type I allergies aims at inducing T-cell immunity while avoiding cross-linking of pre-existing IgE. DNA-based immunotherapy depends on the recruitment of antigen-specific T(H)1 cells and therefore has to provide the whole repertoire of T-cell epitopes. Ubiquitination offers a general approach for the production of hypoallergenic DNA vaccines.
  
  
  OBJECTIVE
  A DNA-based vaccine encoding the major birch pollen allergen Bet v 1 stably linked to ubiquitin was evaluated for its antiallergic potential in a BALB/c mouse model of allergy.
  
  
  METHODS
  Plasmid DNA was applied to mice before (preventive) or after (therapeutic) sensitization with recombinant Bet v 1. In the preventive setting, mice were exposed to aerosolized allergen in addition. Cytokine production was monitored via ELISPOT and Luminex. IgG(1), IgG(2a), and IgE subclass antibody titers were determined by ELISA. In vitro antigen-specific cross-linking of IgE was measured in a degranulation assay. Bronchoalveolar lavages were analyzed for leukocyte subsets as well as for IFN-gamma and IL-5, and paraffin sections of lungs were examined for mucus production and endothelial damage.
  
  
  RESULTS
  Prevaccination with ubiquitinated Bet v 1-stimulated T(H)1-biased immune responses with concomitant suppression of functional IgE, reduction of eosinophil counts in bronchoalveolar lavages, and alleviation of lung pathology, and could also suppress an ongoing IgE response in a therapeutic setting.
  
  
  CONCLUSIONS
  The data clearly demonstrate that hypoallergenic DNA vaccines encoding ubiquitin fusion constructs induce effective antiallergic immune responses.
  
  
  CONCLUSIONS
  Ubiquitination of allergen gene vaccines eliminates the risk of IgE cross-linking, thereby meeting the safety requirements for clinical applications.
authors.affiliation: Department of Molecular Biology, Division of Allergy and Immunology, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.
authors.forename: Roman
authors.initials: R
authors.lastname: Bauer
authors.forename: Sandra
authors.initials: S
authors.lastname: Scheiblhofer
authors.forename: Kerstin
authors.initials: K
authors.lastname: Kern
authors.forename: Christina
authors.initials: C
authors.lastname: Gruber
authors.forename: Tatjana
authors.initials: T
authors.lastname: Stepanoska
authors.forename: Theresa
authors.initials: T
authors.lastname: Thalhamer
authors.forename: Cornelia
authors.initials: C
authors.lastname: Hauser-Kronberger
authors.forename: Beate
authors.initials: B
authors.lastname: Alinger
authors.forename: Thomas
authors.initials: T
authors.lastname: Zoegg
authors.forename: Maximilian
authors.initials: M
authors.lastname: Gabler
authors.forename: Fatima
authors.initials: F
authors.lastname: Ferreira
authors.forename: Arnulf
authors.initials: A
authors.lastname: Hartl
authors.forename: Josef
authors.initials: J
authors.lastname: Thalhamer
authors.forename: Richard
authors.initials: R
authors.lastname: Weiss
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology*; D000818:Animals; D052179:Antigens, Plant; D004195:Disease Models, Animal; D005260:Female; D006967:Hypersensitivity / Q000628:therapy*; D007073:Immunoglobulin E / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D018417:Th1 Cells / Q000276:immunology*; D025801:Ubiquitin / Q000378:metabolism*; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D025801:Ubiquitin; D019444:Vaccines, DNA; D007073:Immunoglobulin E