A recombinant allergen chimer as novel mucosal vaccine candidate for prevention of multi-sensitivities.
BACKGROUND
As conventional immunotherapy is less efficacious in patients with allergic multi-sensitivities compared with mono-sensitized subjects, new intervention strategies are needed. Therefore, an allergen chimer was genetically engineered for treatment of multi-sensitization with birch and grass pollen on the basis of mucosal tolerance induction.


METHODS
The major birch pollen allergen Bet v 1 served as a scaffold for N- and C-terminal linkage of the immunodominant peptides of the grass pollen allergens Phl p 1 and Phl p 5 and this new construct was cloned and expressed in Escherichia coli. After purification, physicochemical and immunological characterization the chimer was used for intranasal tolerance induction prior to poly-sensitization with Bet v 1, Phl p 1 and Phl p 5.


RESULTS
The immunological characterization revealed that the conformation of Bet v 1 within the chimer was comparable to that of natural as well as recombinant Bet v 1. The chimer was immunogenic in mice for T and B cell responses to the three allergens. Intranasal application of the chimer prior to poly-sensitization significantly suppressed humoral and cellular allergen-specific Th2 responses and prevented development of airway inflammation upon allergen challenge. Moreover, local allergen-specific IgA antibodies were induced by the chimer. The mechanisms of poly-tolerance induction seemed to be mediated by regulatory cytokines, since TGF-beta and IL-10 mRNA in splenocytes were upregulated and tolerance was transferable with these cells.


CONCLUSIONS
The data indicate that such allergen chimers harboring several unrelated allergens or allergen peptides could serve as mucosal polyvalent vaccines for prevention of multi-sensitivities.
Wild
C
C
Department of Specific Prophylaxis and Tropical Medicine, Center for Physiology and Pathophysiology, Medical University of Vienna, Vienna, Austria.
Wallner
M
M
Hufnagl
K
K
Fuchs
H
H
Hoffmann-Sommergruber
K
K
Breiteneder
H
H
Scheiner
O
O
Ferreira
F
F
Wiedermann
U
U
D000281:Administration, Intranasal; D000485:Allergens / Q000008:administration & dosage / Q000627:therapeutic use*; D000818:Animals; D029662:Betula / Q000276:immunology; D003888:Desensitization, Immunologic; D005260:Female; D006967:Hypersensitivity / Q000517:prevention & control*; D018928:Immunity, Mucosal; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D006109:Poaceae / Q000276:immunology; D011058:Pollen / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000627:therapeutic use*; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000138:chemical synthesis / Q000627:therapeutic use
D000485:Allergens; D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit