TY - JOUR
T1 - Visualization of clustered IgE epitopes on α-lactalbumin
AU - Hochwallner, Heidrun
AU - Schulmeister, Ulrike
AU - Swoboda, Ines
AU - Focke-Tejkl, Margit
AU - Civaj, Vera
AU - Balic, Nadja
AU - Nystrand, Mats
AU - Härlin, Annika
AU - Thalhamer, Josef
AU - Scheiblhofer, Sandra
AU - Keller, Walter
AU - Pavkov, Tea
AU - Zafred, Domen
AU - Niggemann, Bodo
AU - Quirce, Santiago
AU - Mari, Adriano
AU - Pauli, Gabrielle
AU - Ebner, Christof
AU - Papadopoulos, Nikolaos G.
AU - Herz, Udo
AU - van Tol, Eric A F
AU - Valenta, Rudolf
AU - Spitzauer, Susanne
PY - 2010/6
Y1 - 2010/6
N2 - Background: α-Lactalbumin (α-La) is a major cow's milk (CM) allergen responsible for allergic reactions in infants. Objective: We performed molecular, structural, and immunologic characterization of α-La. Methods: Recombinant α-lactalbumin (rα-La) was expressed in Escherichia coli, purified to homogeneity, and characterized by means of mass spectrometry and circular dichroism, and its allergenic activity was studied by using microarray technology, as well as in a basophil histamine release assay. IgE epitope mapping was performed with synthetic peptides. Results: According to circular dichroism analysis, rα-La represented a folded protein with a high thermal stability and refolding capacity. rα-La reacted with IgE antibodies from 57.6% of patients with CM allergy (n = 66) and induced the strongest basophil degranulation with sera from patients with CM allergy who had exhibited gastrointestinal symptoms or severe systemic reactions on CM exposure. rα-La contained sequential and conformational IgE epitopes. Superposition of IgE-reactive peptides onto the 3-dimensional structure of α-La revealed a close vicinity of the N- and C-terminal peptides within a surface-exposed patch. Conclusions: rα-La can be used for the diagnosis of patients with severe allergic reactions to CM and serves as a paradigmatic tool for the development of therapeutic strategies for CM allergy. © 2010 American Academy of Allergy, Asthma & Immunology.
AB - Background: α-Lactalbumin (α-La) is a major cow's milk (CM) allergen responsible for allergic reactions in infants. Objective: We performed molecular, structural, and immunologic characterization of α-La. Methods: Recombinant α-lactalbumin (rα-La) was expressed in Escherichia coli, purified to homogeneity, and characterized by means of mass spectrometry and circular dichroism, and its allergenic activity was studied by using microarray technology, as well as in a basophil histamine release assay. IgE epitope mapping was performed with synthetic peptides. Results: According to circular dichroism analysis, rα-La represented a folded protein with a high thermal stability and refolding capacity. rα-La reacted with IgE antibodies from 57.6% of patients with CM allergy (n = 66) and induced the strongest basophil degranulation with sera from patients with CM allergy who had exhibited gastrointestinal symptoms or severe systemic reactions on CM exposure. rα-La contained sequential and conformational IgE epitopes. Superposition of IgE-reactive peptides onto the 3-dimensional structure of α-La revealed a close vicinity of the N- and C-terminal peptides within a surface-exposed patch. Conclusions: rα-La can be used for the diagnosis of patients with severe allergic reactions to CM and serves as a paradigmatic tool for the development of therapeutic strategies for CM allergy. © 2010 American Academy of Allergy, Asthma & Immunology.
U2 - 10.1016/j.jaci.2010.03.007
DO - 10.1016/j.jaci.2010.03.007
M3 - Article
C2 - 20466413
SN - 0091-6749
VL - 125
SP - 1279-e9
JO - Journal of Allergy and Clinical Immunology
JF - Journal of Allergy and Clinical Immunology
IS - 6
ER -
