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  • Open Access

Hypoallergenic allergen derivatives of Pru p 3 for immunotherapy of IgE-mediated peach allergy

  • 1,
  • 1,
  • 2,
  • 1,
  • 3,
  • 4 and
  • 1
Clinical and Translational Allergy20133 (Suppl 3) :P177

https://doi.org/10.1186/2045-7022-3-S3-P177

  • Published:

Keywords

  • Lipid Transfer Protein
  • Specific Immunotherapy
  • Vector pET27b
  • Food Hypersensitivity
  • Expression Vector pET27b

Background

IgE-mediated allergy to peach represents a frequent cause of severe anaphylactic reactions in Mediterranean countries. Pru p 3, a nonspecific lipid transfer protein (LTP) from peach, was identified as an important elicitor of IgE-mediated food hypersensitivity reactions to peach. Specific immunotherapy is currently the only allergen-specific and disease-modifying treatment of allergies, but has so far not been applied to food allergic patients due to the high risk of IgE-mediated side effects. Hypoallergenic Pru p 3 derivatives have therefore been constructed in order to minimize side-effects, but these molecules were not immunogenic in animal models.

Methods

Two recombinant Pru p 3-derivatives were developed based on the Pru p 3 amino acid sequence, by introduction of point mutations, or rearrangements of Pru p 3 fragments and oligomerization. Genes encoding the resulting Pru p 3 derivatives were cloned into the expression vector pET27b and expressed in Escherichia coli. The proteins were purified by affinity chromatography and analyzed by SDS-PAGE and mass spectrometry. IgE-reactivity was tested by dot blot analysis and immunogenicity was investigated by immunization of rabbits.

Results

Expression of the two recombinant Pru p 3 derivatives in Escherichia coli yielded high amounts of recombinant proteins. They were purified to homogeneity and exhibited a strongly reduced IgE-reactivity when exposed to sera of peach allergic patients. Upon immunization of rabbits the Pru p 3 derivatives induced a robust IgG antibody response specific for natural Pru p 3.

Conclusion

The reported recombinant hypoallergenic Pru p 3 derivatives are candidates for specific immunotherapy of peach allergic patients.

Disclosure of interest

B Linhart: None declared, A Gstoettner: None declared, C Gamez: None declared, I Swoboda: None declared, A Mari: None declared, N Papadopoulos: None declared, R Valenta: Grant/research support from Biomay AG, Vienna, Austria.

Declarations

Acknowledgement

This work was supported by a research grant from Biomay AG, Vienna, Austria.

Authors’ Affiliations

(1)
Dept. of Pathophysiology and Allergy Research, Medical University of Vienna, Vienna, Austria
(2)
Institute for Medical Research, Foundation Jimenez Diaz, Madrid, Spain
(3)
Center for Molecular Allergology, IDI-IRCCS, Rome, Italy
(4)
Allergy Research Center, 2nd Pediatric Department, National & Kapodistrian University of Athens, Athens, Greece

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