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Baranyi, U; Gattringer, M; Boehm, A; Marth, K; Focke-Tejkl, M; Bohle, B; Blatt, K; Valent, P; Valenta, R; Wekerle, T.
Expression of a major plant allergen as membrane-anchored and secreted protein in human cells with preserved T cell and B cell epitopes.
Int Arch Allergy Immunol. 2011; 156(3):259-266
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Authors Med Uni Graz:
Baranyi Ulrike
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Abstract:
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.
Find related publications in this database (using NLM MeSH Indexing)
Allergens - biosynthesis Allergens - genetics Allergens - immunology
Antigens, Plant -
Antigens, Surface - biosynthesis Antigens, Surface - genetics Antigens, Surface - immunology
Epitopes, B-Lymphocyte - immunology
Epitopes, T-Lymphocyte - immunology
Genetic Vectors -
HEK293 Cells -
Humans -
Hypersensitivity, Immediate - immunology
Immunoglobulin E - immunology Immunoglobulin E - metabolism
Leukocytes, Mononuclear - immunology Leukocytes, Mononuclear - metabolism
Membrane Proteins - immunology
Plant Proteins - biosynthesis Plant Proteins - genetics Plant Proteins - immunology
Plants - immunology Plants - metabolism
Poaceae - immunology
Pollen - chemistry Pollen - immunology Pollen - metabolism
Ribonucleases - biosynthesis Ribonucleases - genetics Ribonucleases - immunology
Transfection -

Find related publications in this database (Keywords)
IgE-mediated allergy
Recombinant allergen Phl p 5
Cell-based therapy
Allergenic activity
Surface protein expression
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