Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties

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Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties Rodrigo Barderas, BS, Mayte Villalba, PhD, Cristina Y. Pascual, MD, PhD, Eva Batanero, PhD, Rosalía Rodríguez, PhD Journal of Allergy and Clinical Immunology Volume 113, Issue 6, Pages 1192-1198 (June 2004) DOI: 10.1016/j.jaci.2003.12.587 Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Antigenic reactivity of chenopod pollen. A, Chenopod-specific IgE levels and antigen-specific IgE values for 104 sera from patients with chenopod allergy. B, Analysis of IgE reactivity for 12 of these sera and for a pool of nonallergic sera (lane N). C, Analysis of IgG reactivity against Ole e 2 (olive profilin)– and Ole e 3 (olive polcalcin)–specific polyclonal antisera (lanes α-Ole e 2 and α-Ole e 3, respectively). Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 SDS-PAGE analysis of purification steps of Che a 2 and Che a 3. A, Protein staining with AgNO3. B, Immunoblotting of proteins assayed with polcalcin- and profilin-specific polyclonal antisera (lanes pAb) or with a pool of sera from patients with chenopod allergy (lanes Sera). Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 Inhibition of the IgE binding of sera from patients with chenopod allergy to the whole pollen extract by the purified Che a 2 and Che a 3. Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 Cross-reactivity between chenopod and olive profilins and chenopod and olive polcalcins analyzed by means of immunoblotting. Strips with purified allergens (left panel, Che a 2; right panel, Che a 3) were incubated with a pool of allergic sera preabsorbed with BSA or the corresponding panallergen from chenopod and olive as inhibitors. Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 Nucleotide sequence of a cDNA encoding Che a 2 and its deduced amino acid sequence (A): the primers used in the cloning are underlined. Comparison of amino acid sequence of Che a 2 with allergenic profilins from other plant sources (B): percentage of identity (%I) and similarity (%S) are referred to the Che a 2.1 clone, and secondary structure of birch profilin23 is indicated. H, α-Helix; black arrow, β-sheet. Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 6 Nucleotide sequence of a cDNA encoding Che a 3 and its deduced amino acid sequence (A): 3 primers used are underlined. Comparison of amino acid sequence of Che a 3 with other allergenic polcalcins (B): percentage of identity (%I) and similarity (%S) are referred to Che a 3, and secondary structure of timothy polcalcin is indicated.32 H, α-Helix; black arrow, β-sheet. Journal of Allergy and Clinical Immunology 2004 113, 1192-1198DOI: (10.1016/j.jaci.2003.12.587) Copyright © 2004 American Academy of Allergy, Asthma and Immunology Terms and Conditions