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Biologic allergen assay for in vivo test allergens with an in vitro model of the murine type I reaction  Andreas Hoffmann, VMD, Stefan Vieths, PhD, Dieter.

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Presentation on theme: "Biologic allergen assay for in vivo test allergens with an in vitro model of the murine type I reaction  Andreas Hoffmann, VMD, Stefan Vieths, PhD, Dieter."— Presentation transcript:

1 Biologic allergen assay for in vivo test allergens with an in vitro model of the murine type I reaction  Andreas Hoffmann, VMD, Stefan Vieths, PhD, Dieter Haustein, PhD  Journal of Allergy and Clinical Immunology  Volume 99, Issue 2, Pages (February 1997) DOI: /S (97) Copyright © 1997 Mosby, Inc. Terms and Conditions

2 FIG. 1 Cross-reactivity between prick test solutions from birch, alder, and hazel pollen. RBL cells were sensitized with IgE from antisera raised against birch pollen. β-Hexosaminidase release was detected after stimulation with tree pollen prick test solution of birch (filled triangles), alder (filled circles), and hazel pollen (filled squares). From each species two batches (dotted or continuous line) were compared. The declared activity of all solutions was 10 HEP. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

3 FIG. 2 Cross-reactivity between isolated allergens or extracts from birch pollen, apple, or celery. RBL cells were sensitized with IgE from antisera raised against birch pollen. Degranulation was measured by β-hexosaminidase release after stimulation with extract from apple (filled circles, continuous line), the apple major allergen Mal d 1 (filled circles, dotted line), and celery (filled squares, dotted line). For comparison the closely related release triggered by natural Bet v 1 (filled triangles, continuous line) and recombinant Bet v 1 (filled triangles, dotted line) is shown. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

4 FIG. 3 Comparison of Fel d 1 concentration of extracts determined by ELISA or RBL assay. The Fel d 1 content from seven cat dander test solutions (filled circles) and 10 cat hair extracts (asterisk) was measured by two-site binding ELISA. The biologic activity detected by RBL assay is also expressed in mass units of Fel d 1 with the same reference. A log/log correlation (r = 0.93) was calculated. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

5 FIG. 4 Comparison of standardized diagnostic allergen solutions of different strengths by RBL assay. A, Cat dander extracts. RBL cells were sensitized with IgE from antisera raised against cat dander. β-Hexosaminidase release was determined after stimulation by serial dilutions of cat dander extracts with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). Fel d 1 (asterisk) reference (50 μg/ml). B, Birch pollen extracts. RBL cells were sensitized with anti-birch pollen serum. Stimulation by birch pollen test solutions with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), or 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). For reference, recombinant Bet v 1 (asterisk), 1 mg/ml, was used. C, Bee venom. RBL cells were sensitized with anti-PLA 2 serum. Mediator release was induced by incubation with serial dilutions of bee venom prick test solutions with a declared content of 300 μg (filled triangles), 100 μg (filled squares), and 10 μg (filled circles) per milliliter. As a reference, we used bee venom PLA 2 (asterisk), 1 mg/ml. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

6 FIG. 4 Comparison of standardized diagnostic allergen solutions of different strengths by RBL assay. A, Cat dander extracts. RBL cells were sensitized with IgE from antisera raised against cat dander. β-Hexosaminidase release was determined after stimulation by serial dilutions of cat dander extracts with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). Fel d 1 (asterisk) reference (50 μg/ml). B, Birch pollen extracts. RBL cells were sensitized with anti-birch pollen serum. Stimulation by birch pollen test solutions with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), or 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). For reference, recombinant Bet v 1 (asterisk), 1 mg/ml, was used. C, Bee venom. RBL cells were sensitized with anti-PLA 2 serum. Mediator release was induced by incubation with serial dilutions of bee venom prick test solutions with a declared content of 300 μg (filled triangles), 100 μg (filled squares), and 10 μg (filled circles) per milliliter. As a reference, we used bee venom PLA 2 (asterisk), 1 mg/ml. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

7 FIG. 4 Comparison of standardized diagnostic allergen solutions of different strengths by RBL assay. A, Cat dander extracts. RBL cells were sensitized with IgE from antisera raised against cat dander. β-Hexosaminidase release was determined after stimulation by serial dilutions of cat dander extracts with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). Fel d 1 (asterisk) reference (50 μg/ml). B, Birch pollen extracts. RBL cells were sensitized with anti-birch pollen serum. Stimulation by birch pollen test solutions with a labeled amount of 10 HEP (filled triangles), 100,000 SQ-U (filled squares), or 1000 SQ-U (filled circles) from each of two batches (dotted or continuous line). For reference, recombinant Bet v 1 (asterisk), 1 mg/ml, was used. C, Bee venom. RBL cells were sensitized with anti-PLA 2 serum. Mediator release was induced by incubation with serial dilutions of bee venom prick test solutions with a declared content of 300 μg (filled triangles), 100 μg (filled squares), and 10 μg (filled circles) per milliliter. As a reference, we used bee venom PLA 2 (asterisk), 1 mg/ml. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

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