The Journal of Allergy and Clinical Immunology
Volume 120, Issue 2 , Pages 374-380, August 2007

Allergy multivaccines created by DNA shuffling of tree pollen allergens

  • Michael Wallner, PhD

      Affiliations

    • From the Christian Doppler Laboratory of Allergy Diagnosis and Therapy
    • ,
  • Angelika Stöcklinger, MSc

      Affiliations

    • From the Christian Doppler Laboratory of Allergy Diagnosis and Therapy
    • ,
  • Theresa Thalhamer, MSc

      Affiliations

    • Department of Molecular Biology, University of Salzburg
    • ,
  • Barbara Bohle, PhD

      Affiliations

    • Department of Pathophysiology, Center of Physiology and Pathophysiology, Medical University of Vienna
    • ,
  • Lothar Vogel, PhD

      Affiliations

    • Division of Allergology, Paul-Ehrlich-Institut, Langen
    • ,
  • Peter Briza, PhD

      Affiliations

    • Department of Molecular Biology, University of Salzburg
    • ,
  • Heimo Breiteneder, PhD

      Affiliations

    • Department of Pathophysiology, Center of Physiology and Pathophysiology, Medical University of Vienna
    • ,
  • Stefan Vieths, PhD

      Affiliations

    • Division of Allergology, Paul-Ehrlich-Institut, Langen
    • ,
  • Arnulf Hartl, PhD

      Affiliations

    • Paracelsus Private Medical University, Salzburg
    • ,
  • Adriano Mari, MD

      Affiliations

    • Center for Clinical and Experimental Allergology, IDI-IRCCS, Rome
    • ,
  • Christof Ebner, MD

      Affiliations

    • Allergieambulatorium, Vienna
    • ,
  • Peter Lackner, PhD

      Affiliations

    • Department of Molecular Biology, University of Salzburg
    • ,
  • Peter Hammerl, PhD

      Affiliations

    • From the Christian Doppler Laboratory of Allergy Diagnosis and Therapy
    • ,
  • Josef Thalhamer, PhD

      Affiliations

    • From the Christian Doppler Laboratory of Allergy Diagnosis and Therapy
    • ,
  • Fatima Ferreira, PhD

      Affiliations

    • From the Christian Doppler Laboratory of Allergy Diagnosis and Therapy
    • Corresponding Author InformationReprint requests: Fatima Ferreira, PhD, Department of Molecular Biology, University of Salzburg, Hellbrunnerstr. 34, A-5020 Salzburg, Austria.

Received 16 February 2007; received in revised form 20 April 2007; accepted 16 May 2007. published online 12 July 2007.

Salzburg and Vienna, Austria, Langen, Germany, and Rome, Italy

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.

Conclusion

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.

Clinical implications

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.

Key words: Gene shuffling, tree pollen allergens, Bet v 1 family, T-cell epitope, IgE epitope, IgE cross-reactivity, genetic immunization, hypoallergen, allergen chimera

Abbreviations used: ELISpot, Enzyme-linked immunosorbent spot, RBL, Rat basophilic leukemia, SIT, Allergen-specific immunotherapy, TCL, T-cell line, TPC, Tree pollen chimera

 

 Supported by “Fonds zur Förderung der Wissenschaftlichen Forschung, FWF” Vienna, Austria, grants P16456-B05, NFN S88 (S8811, S8813), and FSB18 (F1802, F1807).Disclosure of potential conflict of interest: L. Vogel and S. Vieths have received grants/research support from the European Union (EuroPrevall). S. Vieths is also an associate of the Institute for Product Quality in Berlin, Germany, and has received grants/research support from the German Research Society, the Research Fund of the German Food Industry, and Monsanto.

PII: S0091-6749(07)00987-6

doi:10.1016/j.jaci.2007.05.021

The Journal of Allergy and Clinical Immunology
Volume 120, Issue 2 , Pages 374-380, August 2007

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