Induction of IgE antibodies in mice and rhesus monkeys with recombinant birch pollen allergens: Different allergenicity of Bet v 1 and Bet v 2 Susanne.

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Presentation transcript:

Induction of IgE antibodies in mice and rhesus monkeys with recombinant birch pollen allergens: Different allergenicity of Bet v 1 and Bet v 2 Susanne Vrtala, PhDa, Peter Mayer, VMDb, Fatima Ferreira, PhDc, Markus Susani, PhDd, Alec H. Sehon, PhDe, Dietrich Kraft, MDa, Rudolf Valenta, MDa Journal of Allergy and Clinical Immunology Volume 98, Issue 5, Pages 913-921 (November 1996) DOI: 10.1016/S0091-6749(96)80007-8 Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 1 Coomassie Brilliant Blue–stained SDS-PAGE showing purified recombinant Bet v 1 and recombinant birch profilin (Bet v 2) separated under reducing and nonreducing conditions. Five micrograms of purified recombinant Bet v 1 and 5 mg of purified recombinant Bet v 2 were separated by SDS-PAGE under reducing (+ β-mercaptoethanol) and nonreducing (−β-mercaptoethanol) conditions. Lane M is the molecular weight marker. Monomers (1) and dimers (2) in the nonreduced samples of Bet v 2 are indicated with asterisks. β-Met, β-Mercaptoethanol. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 2 Detection of Bet v 2 polymers with profilin-specific anisera. Recombinant Bet v 2 was separated by SDS-PAGE and blotted onto nitrocellulose under nonreducing conditions and detected with two different anti-profilin antisera (lane 1: RP1; lane 2: RP3). In lane 3 a normal rabbit serum was used as a negative control. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 IgE and lgG reactivity of a representative Bet v 1–immunized rhesus monkey (A) and a Bet v 2–immunized rhesus monkey (B) with nitrocellulose-blotted rBet v 1 and rBet v 2. For IgE detection, sera were diluted 1:10, and for IgG detection, 1:1000. To allow a comparison of signal intensity all blots were exposed for the same time (4 hours). In the first lane the monkeys' preimmune serum was used, whereas in the other lanes, strips were incubated with serum samples obtained 1 week before and after the monthly immunizations. The Bet v 1–immunized monkey had received 1 μg of rBet v 1 per immunization, whereas the Bet v 2–immunized monkey was immunized with 20 μg of rBet v 2 per injection. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 IgE and lgG reactivity of a representative Bet v 1–immunized rhesus monkey (A) and a Bet v 2–immunized rhesus monkey (B) with nitrocellulose-blotted rBet v 1 and rBet v 2. For IgE detection, sera were diluted 1:10, and for IgG detection, 1:1000. To allow a comparison of signal intensity all blots were exposed for the same time (4 hours). In the first lane the monkeys' preimmune serum was used, whereas in the other lanes, strips were incubated with serum samples obtained 1 week before and after the monthly immunizations. The Bet v 1–immunized monkey had received 1 μg of rBet v 1 per immunization, whereas the Bet v 2–immunized monkey was immunized with 20 μg of rBet v 2 per injection. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 IgE and lgG reactivity of a representative Bet v 1–immunized rhesus monkey (A) and a Bet v 2–immunized rhesus monkey (B) with nitrocellulose-blotted rBet v 1 and rBet v 2. For IgE detection, sera were diluted 1:10, and for IgG detection, 1:1000. To allow a comparison of signal intensity all blots were exposed for the same time (4 hours). In the first lane the monkeys' preimmune serum was used, whereas in the other lanes, strips were incubated with serum samples obtained 1 week before and after the monthly immunizations. The Bet v 1–immunized monkey had received 1 μg of rBet v 1 per immunization, whereas the Bet v 2–immunized monkey was immunized with 20 μg of rBet v 2 per injection. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 3 IgE and lgG reactivity of a representative Bet v 1–immunized rhesus monkey (A) and a Bet v 2–immunized rhesus monkey (B) with nitrocellulose-blotted rBet v 1 and rBet v 2. For IgE detection, sera were diluted 1:10, and for IgG detection, 1:1000. To allow a comparison of signal intensity all blots were exposed for the same time (4 hours). In the first lane the monkeys' preimmune serum was used, whereas in the other lanes, strips were incubated with serum samples obtained 1 week before and after the monthly immunizations. The Bet v 1–immunized monkey had received 1 μg of rBet v 1 per immunization, whereas the Bet v 2–immunized monkey was immunized with 20 μg of rBet v 2 per injection. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions

FIG. 4 ELISA measurements of Bet v 1–and Bet v 2–specific IgE antibodies in rhesus monkeys. Levels of Bet v 1– and Bet v 2–specific IgE were measured at serum dilutions of 1:5 in five rBet v 1–immunized rhesus monkeys (1 to 5) and in five rBet v 2–immunized monkeys (6 to 10). Extinctions are displayed as optical densities (OD) on the y axis. Numbers of the animals are displayed on the x axis. Journal of Allergy and Clinical Immunology 1996 98, 913-921DOI: (10.1016/S0091-6749(96)80007-8) Copyright © 1996 Mosby, Inc. Terms and Conditions