Effector activity of peanut allergens: a critical role for Ara h 2, Ara h 6, and their variants.
| Autor: | Porterfield HS; Department of Medicine, University of Colorado at Denver, Aurora, CO 80262, USA., Murray KS, Schlichting DG, Chen X, Hansen KC, Duncan MW, Dreskin SC |
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| Jazyk: | angličtina |
| Zdroj: | Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology [Clin Exp Allergy] 2009 Jul; Vol. 39 (7), pp. 1099-108. Date of Electronic Publication: 2009 May 11. |
| DOI: | 10.1111/j.1365-2222.2009.03273.x |
| Abstrakt: | Rationale: An important property of allergens is their ability to cross-link IgE and activate mast cells and basophils. The effector activity of peanut allergens has not been well characterized. Methods: Crude extracts of fresh peanut flour were fractionated by gel filtration. Effector function was assayed by measuring degranulation of RBL SX-38 cells sensitized with IgE from individual sera and from pools of sera of peanut-allergic donors. Results: Following gel filtration, 75 +/- 7% of the applied protein and 76 +/- 16% (n=3) of the applied activity (assayed with a pool of 11 sera) were recovered in the resultant fractions. The majority (85 +/- 2%; n=3) of the recovered activity resided in a fraction with a theoretical average molecular weight of approximately 20 kDa and a range of 13-25 kDa. When all the individual fractions were recombined, the measured activity was similar to that of the original extract [140 +/- 43% when measured with a pool of serum (n=2) and 66 +/- 7% when measured with individual sera (n=4)]; when all individual fractions excluding the 20 kDa fraction were recombined, the measured activity was only 8 +/- 2% (n=2) of the original extract when assayed with the serum pool and 10 +/- 4% (n=3) when assayed with the individual sera. Two-dimensional gel electrophoresis of this biologically active fraction revealed >60 protein spots. Analysis of 50 of the most prominent spots by matrix-assisted laser-desorption ionization time-of-flight mass spectrometry and of the full mixture by automated tandem mass spectrometry coupled to online capillary liquid chromatography revealed that >97% of the protein mass consisted of Ara h 2.0101, Ara h 2.0201, Ara h 6 isoforms, and variants of these proteins. Conclusions: Ara h 2 and Ara h 6 account for the majority of the effector activity found in a crude peanut extract. |
| Databáze: | MEDLINE |
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