Development and characterization of a recombinant, hypoallergenic, peptide-based vaccine for grass pollen allergy.

Abstract

BACKGROUND:

Grass pollen is one of the most important sources of respiratory allergies worldwide.

OBJECTIVE:

This study describes the development of a grass pollen allergy vaccine based on recombinant hypoallergenic derivatives of the major timothy grass pollen allergens Phl p 1, Phl p 2, Phl p 5, and Phl p 6 by using a peptide-carrier approach.

METHODS:

Fusion proteins consisting of nonallergenic peptides from the 4 major timothy grass pollen allergens and the PreS protein from hepatitis B virus as a carrier were expressed in Escherichia coli and purified by means of chromatography. Recombinant PreS fusion proteins were tested for allergenic activity and T-cell activation by means of IgE serology, basophil activation testing, T-cell proliferation assays, and xMAP Luminex technology in patients with grass pollen allergy. Rabbits were immunized with PreS fusion proteins to characterize their immunogenicity.

RESULTS:

Ten hypoallergenic PreS fusion proteins were constructed, expressed, and purified. According to immunogenicity and induction of allergen-specific blocking IgG antibodies, 4 hypoallergenic fusion proteins (BM321, BM322, BM325, and BM326) representing Phl p 1, Phl p 2, Phl p 5, and Phl p 6 were included as components in the vaccine termed BM32. BM321, BM322, BM325, and BM326 showed almost completely abolished allergenic activity and induced significantly reduced T-cell proliferation and release of proinflammatory cytokines in patients' PBMCs compared with grass pollen allergens. On immunization, they induced allergen-specific IgG antibodies, which inhibited patients' IgE binding to all 4 major allergens of grass pollen, as well as allergen-induced basophil activation.

CONCLUSION:

A recombinant hypoallergenic grass pollen allergy vaccine (BM32) consisting of 4 recombinant PreS-fused grass pollen allergen peptides was developed for safe immunotherapy of grass pollen allergy.

• Authors: Margarete Focke-Tejkl, Milena Weber, Katarzyna Niespodziana, Angela Neubauer, Hans Huber, Rainer Henning, Gottfried Stegfellner, Bernhard Maderegger, Martina Hauer, Frank Stolz, Verena Niederberger, Katharina Marth, Julia Eckl-Dorna, Richard Weiss, Josef Thalhamer, Katharina Blatt, Peter Valent, Rudolf Valenta
• Journal: The Journal of allergy and clinical immunology (J Allergy Clin Immunol) Vol. 135 Issue 5 Pg. 1207-7.e1-11 (May 2015) ISSN: 1097-6825 [Electronic] United States
• PMID: 25441634 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

Chemical References

• Allergens
• Cytokines
• Hepatitis B Surface Antigens
• Immunoglobulin G
• Inflammation Mediators
• Peptides
• Recombinant Fusion Proteins
• Vaccines, Subunit
• Vaccines, Synthetic
• Immunoglobulin E

Topics

• Allergens (immunology)
• Animals
• Basophils (immunology, metabolism)
• Cytokines (biosynthesis)
• Disease Models, Animal
• Female
• Hepatitis B Surface Antigens (chemistry, genetics)
• Humans
• Immunoglobulin E (blood, immunology)
• Immunoglobulin G (immunology)
• Inflammation Mediators (metabolism)
• Lymphocyte Activation (immunology)
• Lymphocytes (immunology, metabolism)
• Mice
• Peptides (immunology)
• Poaceae
• Pollen (immunology)
• Protein Binding
• Protein Engineering
• Recombinant Fusion Proteins (chemistry, genetics, immunology, isolation & purification)
• Rhinitis, Allergic, Seasonal (prevention & control)
• Vaccines, Subunit (immunology)
• Vaccines, Synthetic (immunology)