T cell epitope-containing hypoallergenic recombinant fragments of the major birch pollen allergen, Bet v 1, induce blocking antibodies

J Immunol. 2000 Dec 1;165(11):6653-9. doi: 10.4049/jimmunol.165.11.6653.

Abstract

Allergen-specific immunotherapy represents one of the few curative approaches toward type I allergy. Up to 25% of allergic patients are sensitized against the major birch pollen allergen, Bet v 1. By genetic engineering we produced two recombinant (r) Bet v 1 fragments comprising aa 1-74 and aa 75-160 of Bet v 1, which, due to a loss of their native-like fold, failed to bind IgE Abs and had reduced allergenic activity. Here we show that both fragments covering the full Bet v 1 sequence induced human lymphoproliferative responses similar to rBet v 1 wild type. The C-terminal rBet v 1 fragment induced higher lymphoproliferative responses than the N-terminal fragment and represented a Th1-stimulating segment with high IFN-gamma production, whereas the N-terminal fragment induced higher IL-4, IL-5, and IL-13 secretion. Immunization of mice and rabbits with rBet v 1 fragments induced IgG Abs, which cross-reacted with complete Bet v 1 and Bet v 1-related plant allergens and strongly inhibited the IgE binding of allergic patients to these allergens. Thus, our results demonstrate that hypoallergenic T cell epitope-containing rBet v 1 fragments, despite lacking IgE epitopes, can induce Abs in vivo that prevent the IgE binding of allergic patients to the wild-type allergen. The overall demonstration of the immunogenic features of the hypoallergenic rBet v 1 fragments will now enable clinical studies for safer and more efficient specific immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / genetics
  • Allergens / immunology*
  • Allergens / metabolism
  • Animals
  • Antibodies, Blocking / biosynthesis*
  • Antibodies, Blocking / metabolism
  • Antigens, Plant
  • Binding Sites, Antibody / genetics
  • Binding, Competitive / genetics
  • Binding, Competitive / immunology
  • Cells, Cultured
  • Cross Reactions
  • Cytokines / metabolism
  • Desensitization, Immunologic
  • Epitopes, T-Lymphocyte / administration & dosage
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology*
  • Female
  • Humans
  • Immunoglobulin E / metabolism
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / blood
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / metabolism
  • Lymphocyte Activation / genetics
  • Lymphocyte Activation / immunology
  • Mice
  • Mice, Inbred BALB C
  • Peptide Fragments / administration & dosage
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology*
  • Plant Proteins / genetics
  • Plant Proteins / immunology*
  • Plant Proteins / metabolism
  • Pollen / immunology
  • Rabbits
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / immunology*
  • Rhinitis, Allergic, Seasonal / immunology
  • Sequence Homology, Amino Acid
  • Trees

Substances

  • Allergens
  • Antibodies, Blocking
  • Antigens, Plant
  • Cytokines
  • Epitopes, T-Lymphocyte
  • Immunoglobulin G
  • Peptide Fragments
  • Plant Proteins
  • Recombinant Proteins
  • Bet v 1 allergen, Betula
  • Immunoglobulin E