Intranasal Treatment with a Recombinant Hypoallergenic Derivative of the Major Birch Pollen Allergen Bet v 1 Prevents Allergic Sensitization and Airway Inflammation in Mice

Wiedermann, Ursula; Herz, Udo; Baier, K.; Vrtala, Susanne; Neuhaus-Steinmetz, Ulrich; Bohle, Barbara; Dekan, Gerhard; Renz, Harald; Ebner, Christof; Valenta, Rudolf; Kraft, Dietrich

doi:10.1159/000049496

Cited by 52 publications

(63 citation statements)

References 34 publications

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“…In comparison to spleen cells from naive mice ( n = 13) stimulated with rBet v 1 (SI = 1·12 ± 0·11 SEM), proliferative responses of mice from all three experimental groups could be considered significant ( P < 0·05). No differences between the groups were found in epitope mapping experiments, revealing a major T cell epitope at the C terminus of the Bet v 1 molecule as previously described [22,24].…”

Section: Allergen Specific Stimulation Of Splenocytes Proliferative Rsupporting

confidence: 72%

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Repa

Wild²,

Hufnagl³

et al. 2004

Clinical and Experimental Immunology

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SUMMARYThe pathophysiological and immunological characteristics of allergic immune responses are controlled by a variety of factors. We have studied the extent to which the route of sensitization influences allergenspecific IgE synthesis and local airway inflammation using a mouse model of allergic sensitization to the major birch pollen allergen Bet v 1. Sensitization of BALB/c mice with recombinant (r)Bet v 1 was performed using intraperitoneal (IP), subcutaneous (SC) or aerosol (AS) sensitization protocols. Mice were analysed for allergen-specific serum antibodies by ELISA and IgE-dependent basophil degranulation. Proliferative responses and cytokine production of splenocytes were measured upon Bet v 1 stimulation in vitro . Bronchoalveolar lavages were performed after airway challenge with aerosolized birch pollen extract for assessment of eosinophilic airway inflammation and local cytokine production in vivo. Highest allergen specific IgE levels and IgE-dependent basophil degranulation were achieved using the SC route. High IL-5 production by spleen and lung cells was associated with pronounced eosinophilia in bronchoalveolar lavages. After IP sensitization, despite giving the highest IgG levels, only low IgE levels, basophil degranulation and IL-5 production were seen. On the other hand, AS sensitization, resulting in the lowest systemic IgE and IL-5 levels, led to a comparably strong airway inflammation as the SC route. Our finding that the route of sensitization can result in a dissociation of local and systemic immune responses may contribute to a better understanding of the pathogenesis of allergic diseases and help to develop new treatment strategies.Keywords type I allergy murine model sensitization route local immune response systemic immune response

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Section: Allergen Specific Stimulation Of Splenocytes Proliferative Rsupporting

confidence: 72%

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Repa

Wild²,

Hufnagl³

et al. 2004

Clinical and Experimental Immunology

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“…Although the immunological mechanisms of SIT are not fully understood, successful SIT requires preserved T-cell epitopes of the allergens. Therefore, low IgE-binding proteins, such as allergen mutants or fragments with retained T-cell epitopes, are interesting candidates for vaccines for a new strategy of specific immunotherapy with reduced anaphylactic side effects [15,48], and residues involved in IgE binding are obviously promising targets for mutation to generate such hypo-allergenic variants. However, results of tests in vivo with hypo-allergenic variants of food allergens have not been published so far.…”

Section: Discussionmentioning

confidence: 99%

“…In addition to those 2174 NOE distance, 68 backbone torsion angle, and 68 hydrogen bond distance restraints used for the structure calculation of Pru av 1 wt [7], which did not involve Glu 45 , Gly 46 , Asp 47 , Gly 48 , Gly 51 , Thr 52 or Ile 53 , another 147 distance restraints could be derived from the 2D and 3D NOESY spectra in an iterative procedure and another 3 backbone torsion angle restraints from the 3 J HNHα scalar coupling constants measured. These experimental restraints served as an input for the calculation of 60 structures, as described previously [7].…”

Section: Structure Calculationmentioning

confidence: 99%

Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure

Neudecker

Lehmann

Nerkamp

et al. 2003

Biochemical Journal

110

106

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Birch pollinosis is often accompanied by adverse reactions to food due to pollen-allergen specific IgE cross-reacting with homologous food allergens. The tertiary structure of Pru av 1, the major cherry (Prunus avium) allergen, for example, is nearly identical with Bet v 1, the major birch (Betula verrucosa) pollen allergen. In order to define cross-reactive IgE epitopes, we generated and analysed mutants of Pru av 1 and Api g 1.0101, the major celery (Apium graveolens) allergen, by immunoblotting, EAST (enzyme allergosorbent test), CD and NMR spectroscopy. The mutation of Glu 45 to Trp 45 in the P-loop region, a known IgE epitope of Bet v 1, significantly reduced IgE binding to Pru av 1 in a subgroup of cherry-allergic patients. The backbone conformation of Pru av 1 wild-type is conserved in the three-dimensional structure of Pru av 1 Trp 45 , demonstrating that the side chain of Glu 45 is involved in a cross-reactive IgE epitope. Accordingly, for a subgroup of celery-allergic patients, IgE binding to the homologous celery allergen Api g 1.0101 was enhanced by the mutation of Lys 44 to Glu. The almost complete loss of IgE reactivity to the Pru av 1 Pro 112 mutant is due to disruption of its tertiary structure. Neither the mutation Ala 112 nor deletion of the Cterminal residues 155-159 influenced IgE binding to Pru av 1. In conclusion, the structure of the P-loop partially explains the crossreactivity pattern, and modulation of IgE-binding by site-directed mutagenesis is a promising approach to develop hypo-allergenic variants for patient-tailored specific immunotherapy.

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“…Allergen-specific IgE levels in the sera were quantified by the degranulation of rat basophil leukemia (RBL-2H3) cells as previously described. 40 RBL-2H3 cells were plated in 96-well tissue culture plates (4310 4 cells per well) and passively sensitized by incubation with mouse sera at a final dilution of 1 : 30 for 2 h. After washing, Bet v 1 (0.3 mg/ml) was added for 30 min at 37 uC to induce degranulation. Supernatants were incubated with 4-methylumbelliferyl-N-acetyl-b-D-glucosaminide (Sigma-Aldrich, Saint-Louis, MO, USA) for analysis of bhexosaminidase using an Infinite M200 (Tecan Group, Männedorf, Switzerland) fluorescence microplate reader (l ex : 360 nm/l em : 465 nm).…”

Section: Western Blot Analysis Of Zo-1 and Occludinmentioning

confidence: 99%

Colonization of germ-free mice with a mixture of three lactobacillus strains enhances the integrity of gut mucosa and ameliorates allergic sensitization

et al. 2015

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Increasing numbers of clinical trials and animal experiments have shown that probiotic bacteria are promising tools for allergy prevention. Here, we analyzed the immunomodulatory properties of three selected lactobacillus strains and the impact of their mixture on allergic sensitization to Bet v 1 using a gnotobiotic mouse model. We showed that Lactobacillus (L.) rhamnosus LOCK0900, L. rhamnosus LOCK0908 and L. casei LOCK0919 are recognized via Toll-like receptor 2 (TLR2) and nucleotide-binding oligomerization domain-containing protein 2 (NOD2) receptors and stimulate bone marrow-derived dendritic cells to produce cytokines in species-and strain-dependent manners. Colonization of germ-free (GF) mice with a mixture of all three strains (Lmix) improved the intestinal barrier by strengthening the apical junctional complexes of enterocytes and restoring the structures of microfilaments extending into the terminal web. Mice colonized with Lmix and sensitized to the Bet v 1 allergen showed significantly lower levels of allergen-specific IgE, IgG1 and IgG2a and an elevated total IgA level in the sera and intestinal lavages as well as an increased transforming growth factor (TGF)-b level compared with the sensitized GF mice. Splenocytes and mesenteric lymph node cells from the Lmix-colonized mice showed the significant upregulation of TGF-b after in vitro stimulation with Bet v 1. Our results show that Lmix colonization improved the gut epithelial barrier and reduced allergic sensitization to Bet v 1. Furthermore, these findings were accompanied by the increased production of circulating and secretory IgA and the regulatory cytokine TGF-b. Thus, this mixture of three lactobacillus strains shows potential for use in the prevention of increased gut permeability and the onset of allergies in humans. Cellular & Molecular Immunology

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Intranasal Treatment with a Recombinant Hypoallergenic Derivative of the Major Birch Pollen Allergen Bet v 1 Prevents Allergic Sensitization and Airway Inflammation in Mice

Cited by 52 publications

References 34 publications

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy

Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure

Colonization of germ-free mice with a mixture of three lactobacillus strains enhances the integrity of gut mucosa and ameliorates allergic sensitization

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