Immunoreactivity of allergen (Bet v 1) conjugated to crystalline bacterial cell surface layers (S-layers)

Immunotechnology. 1996 Jun;2(2):103-13. doi: 10.1016/1380-2933(96)00041-3.

Abstract

Background: Crystalline cell surface layers (S-layers) from Gram-positive eubacteria had been demonstrated as carrier/adjuvants for chemically synthesized tumor-associated oligosaccharide haptens and capsular polysaccharide antigens of Streptococcus pneumoniae strains.

Objectives: The applicability of S-layers as vaccine carrier for treatment of Type I allergy was investigated.

Study design: Native or cross-linked S-layer self-assembly products and cell wall preparations from Bacillus sphaericus CCM 2177 and Thermoanaerobacter thermohydrosulfuricus L111-69 and L110-69 were used for immobilization of recombinant major birch pollen allergen Bet v 1.

Results and conclusions: Depending on the carrier used, amounts of approximately 20-40 micrograms allergen per mg conjugate could be immobilized. By application of L-glutamic acid dimethyl ester as a spacer, this value could be increased approximately 10-fold. The functionality of the rBet v 1-conjugates was assessed in immunological systems. (i) The presence of intact B-cell epitopes was demonstrated in inhibition experiments using human Bet v 1-specific IgE. (ii) The rBet v 1-S-layer conjugates were immunogenic in mice. (iii) The proliferation of rBet v 1-specific T-cell clones suggested that the peptides created by processing of immobilized Bet v 1 were similar to those derived from natural allergen. (iv) Stimulation of human allergen-specific TH2 lymphocytes with S-layer-conjugated Bet v 1 led to a modulation of the cytokine production pattern from TH2 to TH0/TH1. This study indicates that S-layers may be suitable carriers for few immunotherapeutical vaccines for Type 1 hypersensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adjuvants, Immunologic
  • Allergens / chemistry
  • Allergens / immunology*
  • Animals
  • Antigens, Plant
  • Bacillus / chemistry
  • Bacillus / immunology*
  • Bacterial Proteins / immunology*
  • Cell Wall / immunology
  • Crystallography
  • Cytokines / biosynthesis
  • Epitopes
  • Female
  • Gram-Positive Bacteria / chemistry
  • Gram-Positive Bacteria / immunology
  • Humans
  • Hypersensitivity / therapy
  • Immunoglobulin E / immunology
  • Immunotherapy
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron
  • Plant Proteins / immunology*
  • Pollen / immunology*
  • T-Lymphocytes / immunology
  • Th2 Cells / immunology
  • Trees / immunology
  • Vaccines, Synthetic / chemistry

Substances

  • Adjuvants, Immunologic
  • Allergens
  • Antigens, Plant
  • Bacterial Proteins
  • Cytokines
  • Epitopes
  • Plant Proteins
  • Vaccines, Synthetic
  • Bet v 1 allergen, Betula
  • Immunoglobulin E