Allergy multivaccines created by DNA shuffling of tree pollen allergens.
BACKGROUND
The major allergens of trees belonging to the Fagales order are collectively known as the Bet v 1 family. Members of the Fagales order have distinct geographic distribution, and it is expected that depending on the exposure pattern of the individual, inclusion of other Bet v 1 family members might increase the efficacy of the treatment.


OBJECTIVE
We aimed to generate molecules that are suitable for specific immunotherapy not only against birch pollen allergy but also against allergies caused by other cross-reactive tree pollens.


METHODS
Fourteen genes of the Bet v 1 family were randomly recombined in vitro by means of DNA shuffling. This library of chimeric proteins was screened for molecules displaying low capacity to induce release of inflammatory mediators but with T-cell immunogenicity higher than that of the parental allergens.


RESULTS
Two chimeric proteins were selected from the library of shuffled clones displaying low allergenicity and high immunogenicity, as determined in in vitro assays using human and animal cells and antibodies, as well as in vivo in animal models of allergy.


CONCLUSIONS
Our results show that it is possible to randomly recombine in vitro T- and B-cell epitopes of a family of related allergens and to select chimeric proteins that perfectly match the criteria presently thought to be relevant for improving allergen-specific immunotherapy.


CONCLUSIONS
The hypoallergenic chimeras described here recombine epitopes of the major Fagales pollen allergens and thus can efficiently substitute a mixture of extracts used for treating patients with tree pollen-induced spring pollinosis worldwide.
Wallner
Michael
M
Christian Doppler Laboratory of Allergy Diagnosis and Therapy, University of Salzburg, Austria.
Stöcklinger
Angelika
A
Thalhamer
Theresa
T
Bohle
Barbara
B
Vogel
Lothar
L
Briza
Peter
P
Breiteneder
Heimo
H
Vieths
Stefan
S
Hartl
Arnulf
A
Mari
Adriano
A
Ebner
Christof
C
Lackner
Peter
P
Hammerl
Peter
P
Thalhamer
Josef
J
Ferreira
Fatima
F
D000485:Allergens / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D029662:Betula / Q000276:immunology; D002460:Cell Line; D032621:DNA Shuffling; D018744:DNA, Plant; D000939:Epitopes; D005260:Female; D015723:Gene Library; D006801:Humans; D006967:Hypersensitivity / Q000517:prevention & control*; D007114:Immunization; D007073:Immunoglobulin E / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009000:Monocytes / Q000276:immunology; D011058:Pollen / Q000276:immunology*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014197:Trees / Q000276:immunology*; D017778:Vaccines, Combined / Q000138:chemical synthesis*
D000485:Allergens; D018744:DNA, Plant; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D017778:Vaccines, Combined; D007073:Immunoglobulin E