Lipid transfer protein (Ara h 9) as a new peanut allergen relevant for a Mediterranean allergic population Susanne Krause, PhD, Gerald Reese, PhD, Stefanie.

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Presentation transcript:

Lipid transfer protein (Ara h 9) as a new peanut allergen relevant for a Mediterranean allergic population Susanne Krause, PhD, Gerald Reese, PhD, Stefanie Randow, BTA, Danila Zennaro, MD, Donato Quaratino, MD, Paola Palazzo, BD, Maria Antonietta Ciardiello, PhD, Arnd Petersen, PhD, Wolf-Meinhard Becker, PhD, Adriano Mari, MD Journal of Allergy and Clinical Immunology Volume 124, Issue 4, Pages 771-778.e5 (October 2009) DOI: 10.1016/j.jaci.2009.06.008 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Immunoblot analysis of IgE antibody reactivities of subjects' sera to Ara h 9–enriched peanut extract. No. 1-66, Patients' sera; India ink, protein staining; MW, molecular weight; NC, negative control. One strip (∗subject 32) belongs to group I. Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immunoblot analysis of recombinant Ara h 9.0101 (A) as thioredoxin fusion protein and Ara h 9.0201 (B). Lane 1, India ink staining; lanes 2-6 and 10-14, patients' sera reactive with natural Ara h 9 (numbers in brackets correspond to the lane numbers of Fig 1: lanes 2 [6], 3 [16], 4 [22], 5 [24], 6 [50], 10 [10], 11 [14], 12 [20], 13 [60], 14 [65]); lane 7, anti–His-tag antibody-AP; lanes 8 and 15, negative controls, anti-human IgE-AP; lane 9, negative control, anti-mouse IgG + IgM-AP. Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Alignment of cDNAs of LTPs from cherry (AF221501), peach (AY792996), strawberry (DQ066732), hazelnut (AF329829), and lentil (AY793556). The lentil sequence representing positions 303 to 326 (except position 321) was selected as forward primer LTP_hom_for. The part of the DNA sequences revealing a high sequence identity were used to design a primer (indicated by gray shading). Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

IgE reactivity to basic (A) and acidic peanut extract, fraction below 20 kd (B). Lane I, Protein staining by India ink; lanes 1 to 8, patients' sera of subjects with peanut allergy and specific IgE antibodies to Pru p 3 (the order of patients' sera is identical with that of Fig 1). MW, Molecular weight. Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Optimizing extraction conditions of peanut LTP Optimizing extraction conditions of peanut LTP. Reactivity of rabbit anti–Cor a 8 antiserum against basic peanut extract (A), acidic peanut extract (B), acidic peanut fraction below 20 kd (C), and for comparison, basic (D) and acidic hazelnut extract (E). Lane I, protein staining by India ink; lane 1, buffer control; lane 2, rabbit serum control; lane 3, rabbit anti–Cor a 8; only in B and C, lane 4, human serum control; lane 5, IgE-reactive patient serum to Ara h 9. MW, Molecular weight. Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Amino acid sequence alignment of Ara h 9. 0101 and Ara h 9 Amino acid sequence alignment of Ara h 9.0101 and Ara h 9.0201 with the N-terminus of natural Ara h 9 and amino acid sequences of 18 additional isoforms and variants. The signal peptides are shaded in gray. Journal of Allergy and Clinical Immunology 2009 124, 771-778.e5DOI: (10.1016/j.jaci.2009.06.008) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions