title: Hypoallergenic derivatives of the major birch pollen allergen Bet v 1 obtained by rational sequence reassembly.
abstract: BACKGROUND
  At least 100 million patients suffer from birch pollen allergy.
  
  
  OBJECTIVE
  Rational design of recombinant derivatives of the major birch pollen allergen, Bet v 1, characterized by reduced IgE reactivity, preservation of sequences relevant for the induction of allergen-specific blocking IgG, and maintenance of T-cell epitopes for immunotherapy of birch pollen allergy.
  
  
  METHODS
  Three recombinant mosaic proteins derived from Bet v 1 were generated by reassembly of codon-optimized genes coding for Bet v 1 fragments containing the elements for the induction of allergen-specific blocking IgG antibodies and the major T-cell epitopes. The proteins were expressed in Escherichia coli as recombinant mosaic molecules and compared with the Bet v 1 wild-type protein by chemical and structural methods, regarding IgE-binding and IgG-binding capacity, in basophil activation assays and tested for the in vivo induction of IgG responses.
  
  
  RESULTS
  Three recombinant Bet v 1 (rBet v 1) mosaic proteins with strongly reduced IgE reactivity and allergenic activity were expressed and purified. Immunization with the recombinant hypoallergens induced IgG antibodies that inhibited IgE reactivity of patients with allergy to Bet v 1 comparable to those induced with the rBet v 1 wild-type allergen.
  
  
  CONCLUSIONS
  We report the generation and preclinical characterization of 3 hypoallergenic rBet v 1 derivatives with suitable properties for immunotherapy of birch pollen allergy.
authors.affiliation: Division of Immunopathology, Department of Pathophysiology, Center of Pathophysiology, Infectiology and Immunology, Vienna General Hospital (AKH), Medical University of Vienna, Vienna, Austria.
authors.forename: Raffaela
authors.initials: R
authors.lastname: Campana
authors.forename: Susanne
authors.initials: S
authors.lastname: Vrtala
authors.forename: Bernhard
authors.initials: B
authors.lastname: Maderegger
authors.forename: Peter
authors.initials: P
authors.lastname: Jertschin
authors.forename: Gottfried
authors.initials: G
authors.lastname: Stegfellner
authors.forename: Ines
authors.initials: I
authors.lastname: Swoboda
authors.forename: Margarete
authors.initials: M
authors.lastname: Focke-Tejkl
authors.forename: Katharina
authors.initials: K
authors.lastname: Blatt
authors.forename: Anna
authors.initials: A
authors.lastname: Gieras
authors.forename: Domen
authors.initials: D
authors.lastname: Zafred
authors.forename: Angela
authors.initials: A
authors.lastname: Neubauer
authors.forename: Peter
authors.initials: P
authors.lastname: Valent
authors.forename: Walter
authors.initials: W
authors.lastname: Keller
authors.forename: Susanne
authors.initials: S
authors.lastname: Spitzauer
authors.forename: Rudolf
authors.initials: R
authors.lastname: Valenta
mesh_terms: D000818:Animals; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology*; D029662:Betula / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D010940:Plant Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D011058:Pollen / Q000276:immunology; D011817:Rabbits; D011994:Recombinant Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology
chemical_list: D052179:Antigens, Plant; D018984:Epitopes, T-Lymphocyte; D010940:Plant Proteins; D011994:Recombinant Proteins; C061312:Bet v 1 allergen, Betula