Induction of IgE antibodies with predefined specificity in rhesus monkeys with recombinant birch pollen allergens, Bet v 1 and Bet v 2 Fatima D. Ferreira,

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Presentation transcript:

Induction of IgE antibodies with predefined specificity in rhesus monkeys with recombinant birch pollen allergens, Bet v 1 and Bet v 2 Fatima D. Ferreira, PhDa, Peter Mayer, VMDb, Wolfgang R. Sperr, MDc, Peter Valent, MDc, Susanne Seiberler, MScd, Christof Ebner, PhDd, Ekke Liehl, PhDb, Otto Scheiner, PhDd, Dietrich Kraft, MDd, Rudolf Valenta, MDd Journal of Allergy and Clinical Immunology Volume 97, Issue 1, Pages 95-103 (January 1996) DOI: 10.1016/S0091-6749(96)70287-7 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Allergic monkeys display IgE cross-reactivity with Bet v 1– and Bet v 2–profilin homologous proteins. Serum IgE from two Bet v 1–allergic monkeys (A and B) was used to detect the major hazel pollen allergen, Cor a 1 and the major apple allergen (A). In lane 1 nitrocelluloses were tested with the preimmune sera, whereas in lane 2 immune sera were used. Two of the profilin-allergic monkeys (A and B) were tested for IgE reactivity with timothy grass (Phleum pratense) profilin (B). In lane 1 the preimmune serum was used, and in lane 2 the immune serum was used. Arrows indicate IgE binding to natural allergens. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Allergic monkeys display IgE cross-reactivity with Bet v 1– and Bet v 2–profilin homologous proteins. Serum IgE from two Bet v 1–allergic monkeys (A and B) was used to detect the major hazel pollen allergen, Cor a 1 and the major apple allergen (A). In lane 1 nitrocelluloses were tested with the preimmune sera, whereas in lane 2 immune sera were used. Two of the profilin-allergic monkeys (A and B) were tested for IgE reactivity with timothy grass (Phleum pratense) profilin (B). In lane 1 the preimmune serum was used, and in lane 2 the immune serum was used. Arrows indicate IgE binding to natural allergens. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Blood basophils of Bet v 1 and monkeys allergic to profilin (Bet v 2) release histamine on allergen stimulation. The representative histamine releases demonstrated in three different Bet v 1–allergic monkeys (D18, D9, and C48) and three profilin (Bet v 2)–allergic monkeys (Vi56, Vi32, and Vi62) were dose-dependent. The horizontal axis indicates the concentration of recombinant allergens in micrograms per milliliter, and the vertical axis shows the percentage of histamine release. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Cellular proliferation assay with PBMC from a Bet v 1–allergic monkey (D9) and a birch profilin (Bet v 2)–allergic monkey (Vi56). PBMC were incubated without agonist (PBMC), with interleukin-2 (positive control), and the recombinant allergens (recombinant Bet v 1 or recombinant birch profilin–Bet v 2), respectively. Thymidine incorporation (cpm) after 6 days of culture is displayed on the horizontal axis. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 Cellular proliferation assay with PBMC from a Bet v 1–allergic monkey (D9) and a birch profilin (Bet v 2)–allergic monkey (Vi56). PBMC were incubated without agonist (PBMC), with interleukin-2 (positive control), and the recombinant allergens (recombinant Bet v 1 or recombinant birch profilin–Bet v 2), respectively. Thymidine incorporation (cpm) after 6 days of culture is displayed on the horizontal axis. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 Influence of systemic corticosteroid application on the IgE reactivity to pollen-derived Bet v 1 in rhesus monkeys reimmunized with recombinant Bet v 1 + AI(OH)3. A group of six rhesus monkeys (A through F) was immunized for 6 months in monthly intervals with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 to induce Bet v 1–specific IgE antibodies. The immunization was stopped for approximately 4 months until the Bet v 1–specific IgE antibodies dropped almost beyond the detection limit (lane 1). Two monkeys were then reimmunized with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 without (A, B) and with a parallel intramuscular injection of corticosteroids (Volon A; 40 mg per animal) on the same day (C, D) or 4 days after each immunization (E, F). Lanes 2 through 4 represent blood samples obtained at the second, third, and fourth immunization, respectively. IgE binding to nitrocellulose-blotted natural Bet v 1 is indicated by arrows. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 Influence of systemic corticosteroid application on the IgE reactivity to pollen-derived Bet v 1 in rhesus monkeys reimmunized with recombinant Bet v 1 + AI(OH)3. A group of six rhesus monkeys (A through F) was immunized for 6 months in monthly intervals with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 to induce Bet v 1–specific IgE antibodies. The immunization was stopped for approximately 4 months until the Bet v 1–specific IgE antibodies dropped almost beyond the detection limit (lane 1). Two monkeys were then reimmunized with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 without (A, B) and with a parallel intramuscular injection of corticosteroids (Volon A; 40 mg per animal) on the same day (C, D) or 4 days after each immunization (E, F). Lanes 2 through 4 represent blood samples obtained at the second, third, and fourth immunization, respectively. IgE binding to nitrocellulose-blotted natural Bet v 1 is indicated by arrows. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 Influence of systemic corticosteroid application on the IgE reactivity to pollen-derived Bet v 1 in rhesus monkeys reimmunized with recombinant Bet v 1 + AI(OH)3. A group of six rhesus monkeys (A through F) was immunized for 6 months in monthly intervals with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 to induce Bet v 1–specific IgE antibodies. The immunization was stopped for approximately 4 months until the Bet v 1–specific IgE antibodies dropped almost beyond the detection limit (lane 1). Two monkeys were then reimmunized with 1 μg of recombinant Bet v 1 adsorbed to AI(OH)3 without (A, B) and with a parallel intramuscular injection of corticosteroids (Volon A; 40 mg per animal) on the same day (C, D) or 4 days after each immunization (E, F). Lanes 2 through 4 represent blood samples obtained at the second, third, and fourth immunization, respectively. IgE binding to nitrocellulose-blotted natural Bet v 1 is indicated by arrows. Journal of Allergy and Clinical Immunology 1996 97, 95-103DOI: (10.1016/S0091-6749(96)70287-7) Copyright © 1996 Mosby, Inc. Terms and Conditions