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A major allergen gene-fusion protein for potential usage in allergen-specific immunotherapy  Fatimah Kussebi, MD, Fariba Karamloo, PhD, Claudio Rhyner,

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Presentation on theme: "A major allergen gene-fusion protein for potential usage in allergen-specific immunotherapy  Fatimah Kussebi, MD, Fariba Karamloo, PhD, Claudio Rhyner,"— Presentation transcript:

1 A major allergen gene-fusion protein for potential usage in allergen-specific immunotherapy 
Fatimah Kussebi, MD, Fariba Karamloo, PhD, Claudio Rhyner, PhD, Peter Schmid-Grendelmeier, MD, Maria Salagianni, PhD, Christian Mannhart, Mübeccel Akdis, MD, PhD, Lyudmilla Soldatova, PhD, Zora Markovic-Housley, PhD, Barbara R. von Beust, PhD, Thomas Kündig, MD, David M. Kemeny, PhD, Kurt Blaser, PhD, Reto Crameri, PhD, Cezmi A. Akdis, MD  Journal of Allergy and Clinical Immunology  Volume 115, Issue 2, Pages (February 2005) DOI: /j.jaci Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

2 Fig 1 Characterization of Api m [1/2] fusion protein. A, Coomassie staining of 0.5 μmol/L recombinant Api m 1, Api m 2, and Api m [1/2] fusion protein. B, Binding of 8 different mouse anti–Api m 2 IgG mAbs to Api m 1 (lane 1), Api m 2 (lane 2), and Api m [1/2] (lane 3). C, Dot blot analysis of human anti–Api m 1 IgG4 mAb binding to Api m 1, Api m 2, and Api m [1/2]. D, Api m [1/2] does not induce IgE and IgG2a against native Api m 1. Api m 1–specific IgE and IgG2a were determined by ELISA. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

3 Fig 2 Low IgE binding to Api m [1/2] fusion protein. A, Serum IgE binding to BV, Api m 1, Api m 2, and the Api m [1/2] fusion protein. B, Serum IgE binding (12 different patients allergic to BV) to equimolar amounts of Api m 1, Api m 2, and Api m [1/2] coated on the solid phase by ELISA. There was no inhibition of serum IgE binding to Api m 1 (C) and Api m 2 (D) coated on a solid phase by Api m [1/2]. BSA was used as control. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

4 Fig 3 Reduced allergenic activity of the Api m [1/2] fusion protein. Induction of leukotriene release (A) and histamine release (B) from basophils of 3 patients allergic to BV stimulated with equimolar amounts of Api m 1, Api m 2, Api m [1/2] fusion protein, and BV. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

5 Fig 4 Induction of T-cell proliferation and cytokine production by Api m [1/2]. PBMCs were stimulated with equimolar amounts of Api m 1, Api m 2, and the Api m [1/2] fusion protein. Thymidine incorporation (3[H] TdR) was measured after 5 days. Background proliferation of unstimulated cultures was between 1570 and 3355 cpm. IL-4, IFN-γ, IL-13, IL-10, and IL-5 were determined in parallel culture supernatants by ELISA. Cytokine production of healthy PBMCs was less than 10% and was omitted from the figure. Horizontal bars represent mean values. SI, Stimulation index. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

6 Fig 5 Api m [1/2] retains T-cell epitopes of Api m 1. Api m 1–specific T-cell clones (BuT 6.1 F10 and BuT 6.6 F4) and Api m 1–nonspecific T-cell clones (BuT 5.1 F12 and BuT 7.3 B4) were stimulated with equimolar amounts of Api m 1, Api m 2, and Api m [1/2] fusion protein for 48 hours. 3[H] thymidine incorporation was measured 12 hours after addition. Ag, Antigen. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

7 Fig 6 Api m [1/2] suppresses antibody development against native allergen. Mice (n=6) were pretreated subcutaneously with Api m [1/2] in saline or saline alone as control 14, 10, and 6 days before intraperitoneal immunization with Api m 1/AL(OH)3. Serum Api m 1–specific IgE, IgG1, and IgG2a were determined. ∗P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

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