Premium
Reducing allergenicity by altering allergen fold: a mosaic protein of Phl p 1 for allergy vaccination
Author(s) -
Ball T.,
Linhart B.,
Sonneck K.,
Blatt K.,
Herrmann H.,
Valent P.,
Stoecklinger A.,
Lupinek C.,
Thalhamer J.,
Fedorov A. A.,
Almo S. C.,
Valenta R.
Publication year - 2009
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1111/j.1398-9995.2008.01910.x
Subject(s) - basophil activation , allergen , immunoglobulin e , immunology , allergy , phleum , basophil , antibody , epitope , medicine , biology , agronomy
Background: The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95% of grass‐pollen‐allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy. Methods: A mosaic protein was generated by PCR‐based re‐assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild‐type by circular dichroism analysis, immunoglobulin E (IgE)‐binding capacity, basophil activation assays and enzyme‐linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice. Results: Grass‐pollen‐allergic patients exhibited greater than an 85% reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1‐specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients’ IgE‐binding to the wild‐type allergen and Phl p 1‐induced degranulation of basophils. Conclusion: We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.