Expression of a major plant allergen as membrane-anchored and secreted protein in human cells with preserved T cell and B cell epitopes.
| Autor: | Baranyi U; Division of Transplantation, Department of Surgery, Medical University of Vienna, Vienna, Austria., Gattringer M, Boehm A, Marth K, Focke-Tejkl M, Bohle B, Blatt K, Valent P, Valenta R, Wekerle T |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | International archives of allergy and immunology [Int Arch Allergy Immunol] 2011; Vol. 156 (3), pp. 259-66. Date of Electronic Publication: 2011 Jun 29. |
| DOI: | 10.1159/000323733 |
| Abstrakt: | Background: Expression of allergens in human cells is a prerequisite for the development of antigen-specific cell therapy in IgE-mediated allergy. We developed a strategy how the clinically relevant major grass pollen allergen Phl p 5 can be efficiently secreted or expressed on the surface of human cells with preserved allergenic activity. Methods: The cDNA of Phl p 5 was fused to a leader peptide with or without a transmembrane domain and both constructs were ligated into a mammalian expression vector. Transfection of these plasmids into human cells resulted in a membrane-anchored or secreted version of Phl p 5, respectively, as determined by ELISA or flow cytometric analysis. Results: Both the secreted and membrane-anchored Phl p 5 proteins bound IgE from allergic patients in an immunoblot assay and induced specific histamine release and CD203c upregulation in basophils of grass pollen-allergic patients. Proliferation of peripheral blood mononuclear cells from Phl p 5-allergic individuals was induced upon stimulation with both variants of Phl p 5 expressed in human cells similar to recombinant Phl p 5. Conclusions: Secreted and membrane-anchored Phl p 5 expressed in human cells preserved B cell as well as T cell epitopes and may be used to develop and test various cell-based strategies for allergen-specific immunomodulation and to delineate the tolerance mechanisms involved therein. (Copyright © 2011 S. Karger AG, Basel.) |
| Databáze: | MEDLINE |
| Externí odkaz: |
|