Suppression of antigen-specific T- and B-cell responses by intranasal or oral administration of recombinant Bet v 1, the major birch pollen allergen, in a murine model of type I allergy,☆☆,

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Abstract

Background: Mucosal (nasal or oral) administration of soluble protein antigens induces a state of antigen-specific immunologic hyporesponsiveness. Several studies have shown that induction of mucosal tolerance can prevent the onset or reduce the severity of certain TH1 -mediated experimental autoimmune diseases. Only a few studies describe similar results for type I allergies, which are caused by excessive TH2 cell activities. Objective: We sought to investigate whether mucosal tolerance induction would also be efficient in preventing an allergic type I immune response. Methods: A murine model of inhalative type I allergy, leading to sensitization to birch pollen and its major allergen Bet v 1 in BALB/c mice, was used. Recombinant Bet v 1 was nasally or orally applied in low doses before sensitization. At the time of death, immediate-type skin tests were performed. Blood was taken, and serum was used for measurement of allergen-specific antibodies. Spleen cell cultures were performed to determine cytokine production (IL-4, IL-5, IL-10, and IFN-γ), as well as levels of TGF-β mRNA. Results: Both nasal and oral administration of minimal doses of recombinant Bet v 1 before aerosol sensitization with birch pollen suppressed the allergen-specific antibody production of all isotypes. Consequently, the in vivo type I skin test responses to the allergen were negative in the tolerized, in contrast to the sensitized, group. Moreover, allergen-specific lymphoproliferative responses and cytokine production in vitro (ie, IFN-γ, IL-4, IL-5, and IL-10) were markedly reduced. In contrast, expression of TGF-β mRNA was markedly increased in spleen cells from nasally tolerized animals, indicating regulatory mechanisms for tolerance induction. Conclusion: We conclude from the present study that nasal, as well as oral, administration of recombinant allergen is an effective way to prevent allergen-specific T- and B-cell responses in a TH2 model. (J Allergy Clin Immunol 1999;103:1202-10.)

Section snippets

Animals

Female, 7-week-old BALB/c mice were obtained from Charles River (Sulzfeld, Germany). All experiments were approved by the Animal Experimentation Ethics Committee of the University of Vienna and the Ministry of Science and Research.

Antigens

Recombinant Bet v 1 (rBet v 1) was obtained from Biomay GesmbH (Linz, Austria). BP (Allergon AB, Engelholm, Sweden) was used for the preparation of a BP extract. Fifty grams of BP was extracted in 500 mL of PBS by overnight stirring at 4°C. After centrifugation at 4000

Antibody responses in tolerized versus sensitized mice

Allergen-specific antibody levels in serum were measured by ELISA 6 days after the last aerosol exposure. Mice, treated intranasally 3 times with 10 μg of rBet v 1 (group 1) and mice fed 3 times with 100 μg of rBet v 1 (group 2) before sensitization, displayed significantly lower IgG1, IgG2a, and IgE antibody levels to r Bet v 1 than the sensitized control animals (group 3) (P < .01). In parallel, the antibody production of all isotypes against the natural BP preparation was abrogated in both

DISCUSSION

In this study we demonstrate that mucosal (particularly intranasal) administration of recombinant Bet v 1 suppressed both T and B cell–derived immune responses and thus prevented sensitization to the allergen. This was documented by an abrogation of allergen-specific T-cell responses and cytokine production in vitro, reduced antibody production, and negative type I skin test reactions in vivo.

Oral/mucosal tolerance has been postulated as one promising concept to control diseases mediated by

Acknowledgements

We thank Renate Steiner-Göltl for excellent technical assistance and Gabriel O’Ríordaín for review and correction of the manuscript.

References (37)

  • H Breiteneder et al.

    The gene coding for the major birch pollen allergen, Bet v 1, is highly homologous to a pea disease resistance response gene

    EMBO J

    (1989)
  • E Jarolim et al.

    IgE and IgG antibodies of patients with allergy to birch pollen as tools to define the allergen profile of Betula verrucosa

    Allergy

    (1989)
  • C Ebner et al.

    Identification of multiple T cell epitopes on Bet v 1, the major birch pollen allergen, using specific T cell clones and overlapping peptides

    J Immunol

    (1993)
  • C Ebner et al.

    Nonallergic individuals recognize the same T cell epitopes of Bet v 1, the major birch pollen allergen, as atopic patients

    J Immunol

    (1995)
  • L Bauer et al.

    Modulation of the allergic immune response in BALB/c mice by subcutaneous injection of high doses of the dominant epitope from the major birch pollen allergen Bet v 1

    Clin Exp Immunol

    (1997)
  • U Wiedermann et al.

    Effects of adjuvants on the immune response to allergens in a murine model of allergen inhalation: cholera toxin induces a Th1-like immune response to Bet v 1, the major birch pollen allergen

    Clin Exp Immunol

    (1998)
  • B Metzler et al.

    Mucosal tolerance in a murine model of experimental autoimmune encephalomyelitis

    Ann NY Acad Sci

    (1996)
  • J Tian et al.

    Nasal administration of glutamate decarboxylase (GAD65) peptides induces Th2 responses and prevents murine insulin-dependent diabetes

    J Exp Med

    (1996)
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    Supported by grants from the Austrian Science Foundation (S06704-MED and S7206-MOB), the European Science Foundation, and ALK (Copenhagen, Denmark). Harald Renz is supported by the Deutsche Forschungsgemeinschaft (Re 737/4-4).

    ☆☆

    Reprint requests: Ursula Wiedermann, MD, PhD, Institute of General and Experimental Pathology, University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.

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