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Cloning, expression, and clinical significance of the major allergen from ash pollen, Fra e 1  Rodrigo Barderas, BSc, Ashok Purohit, MD, Ioanna Papanikolaou,

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Presentation on theme: "Cloning, expression, and clinical significance of the major allergen from ash pollen, Fra e 1  Rodrigo Barderas, BSc, Ashok Purohit, MD, Ioanna Papanikolaou,"— Presentation transcript:

1 Cloning, expression, and clinical significance of the major allergen from ash pollen, Fra e 1 
Rodrigo Barderas, BSc, Ashok Purohit, MD, Ioanna Papanikolaou, MD, Rosalía Rodríguez, PhD, Gabrielle Pauli, MD, Mayte Villalba, PhD  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 Primary structure of Fra e 1. A, Fra e –nucleotide sequence and its deduced amino acid sequence. Sequences corresponding to the primers used for cloning are underlined. B, Alignment of the 2 deduced amino acid sequences with allergenic members of the Oleaceae family, Ole e 1 (olive), Syr v 1 (lilac), and Lig v 1 (privet). The glycosylation site is underlined. Accession numbers of the Fra e and Fra e sequences are Q7XAV4 and AY652744, respectively. ∗∗ Accession numbers of the nucleotide sequences here reported are Q7XAV4 and AY The new allergen has been submitted to the World Health Organization/International Union of Immunological Societies Allergen Nomenclature Sub-Committee. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

3 Fig 2 Expression and purification of rFra e 1. A, Time course of the recombinant production was analyzed by Coomassie Blue staining after SDS-PAGE at different times. B, Coomassie Blue staining of the purification steps and ConA staining of purified protein transferred to membranes after SDS-PAGE. M, molecular mass markers. C, MS analysis of rFra e 1. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

4 Fig 3 Immunologic relationships between Fra e 1 and Ole e 1. A, Immunodetection of rFra e 1 with 7 mAbs (F1 to F7) specific to Ole e 1. B, Inhibition of the IgG binding of pAb to rFra e 1 by using rFra e 1 and rOle e 1 as inhibitors (2 μg). C, Inhibition of the IgE binding to ash pollen extract by using rFra e 1 as inhibitor (5 μg). Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

5 Fig 4 IgE reactivity profile to ash pollen extract and purified rFra e 1. A, IgE binding to nitrocellulose-blotted samples after SDS-PAGE with the sera from 11 patients and a nonatopic control (–) is shown. B, rFra e 1–specific IgE levels in OD versus ash pollen specific IgE levels (in kU/L) for individual sera from the selected population. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

6 Fig 5 Histamine release assays. The responses of 7 patients allergic to ash pollen and the nonatopic control patient are shown. 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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