Identification of common allergenic structures in mugwort and ragweed pollen Reinhold Hirschwehr, MDa, Claudia Heppner, MSca, Susanne Spitzauer, MDb,

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Identification of common allergenic structures in mugwort and ragweed pollen Reinhold Hirschwehr, MDa, Claudia Heppner, MSca, Susanne Spitzauer, MDb, Wolfgang R. Sperr, MDc, Peter Valent, MDc, Uwe Bergerd, Friedrich Horak, MDd, Siegfried Jäger, PhDd, Dietrich Kraft, MDa, Rudolf Valenta, MDa Journal of Allergy and Clinical Immunology Volume 101, Issue 2, Pages 196-206 (February 1998) DOI: 10.1016/S0091-6749(98)70384-7 Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 1 Comparison of the IgE-binding components detected by sera from individuals allergic to mugwort pollen in nitrocellulose-blotted mugwort and ragweed pollen extracts. Nitrocellulose strips containing blotted mugwort pollen extract (A) and ragweed pollen extract (B) were incubated with serum IgE from individuals allergic to mugwort (lanes 1 through 16), serum from a nonallergic individual (lane N), and buffer (lane C). Molecular weights are displayed on right side of each panel. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 2 Effect of reducing conditions on the pattern of IgE-binding components in mugwort and ragweed pollen extracts. Mugwort and ragweed pollen extracts were separated by an analytical SDS-PAGE under reducing and nonreducing conditions (lane 1: mugwort, nonreduced; lane 2: ragweed, nonreduced; lane 3: mugwort, reduced; lane 4: ragweed, reduced) and blotted onto nitrocellulose sheets. Identically prepared sheets were incubated with serum IgE from two different patients allergic to mugwort (A and B), and bound IgE was detected with 125I-labeled anti-human IgE antibodies. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 3 IgE immunoblot inhibition experiment. Sera from seven patients allergic to mugwort (lanes 1 through 7 in blocks A through C) were probed with nitrocellulose-blotted mugwort pollen extract (top) and ragweed pollen extract (bottom). In block D, serum from a nonallergic individual (lane 1) and buffer without addition of serum (lane C) was used. Sera from patients allergic to mugwort were preincubated with ragweed pollen extract (block B, top), mugwort pollen extract (block B, bottom), and dog albumin (block C, top and bottom). Sera in block A (top and bottom) were not preadsorbed. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 4 Detection of profilin as a cross-reactive 14 kd allergen in mugwort and ragweed pollen extract. Nitrocellulose strips containing mugwort pollen extract (A) and ragweed pollen extract (B) were probed with three different rabbit anti-profilin antisera (A, B, and C) in lane 1. Lane C represents buffer control without addition of anti-serum. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 5 Identification of profilin as a cross-reactive 14 kd allergen in mugwort and ragweed pollen extract by inhibition with recombinant birch profilin. Sera from four patients allergic to mugwort (blocks A, B, C, and D) were probed for IgE reactivity with nitrocellulose-blotted mugwort pollen extract (top) and ragweed pollen extract (bottom). In lane 1 sera were not preadsorbed, in lane 2 sera were preincubated with recombinant birch profilin (Bet v 2), and in lane 3 sera were preincubated with recombinant Bet v 1. Bound IgE was detected with 125I-labeled anti-human IgE antibodies. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 6 Basophil histamine release. Granulocytes from an individual allergic to mugwort with profilin-specific IgE were incubated with increasing doses of mugwort pollen extract (open circles), ragweed pollen extract (open diamonds), recombinant birch profilin (open triangles), and anti-human IgE antibodies (filled squares). Percentage of histamine release shown on y-axis reflects means of triplicate determinations. Error bars are displayed. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 7 Measurement of nasal air flow after allergen provocation. Nasal air flow in a volunteer allergic to mugwort pollen (A) and a nonallergic individual (B) was measured after application of isotonic sodium chloride solution (blocks A and C) and after application of mugwort pollen extract (block B) and ragweed pollen extract (block D). Nasal flow was recorded in cubic centimeters per second displayed on the y-axis at the pressure shown on the x-axis during expiration and inspiration. CCM/S, Cubic centimeters per second; PR, pressure in pascals. Journal of Allergy and Clinical Immunology 1998 101, 196-206DOI: (10.1016/S0091-6749(98)70384-7) Copyright © 1998 Mosby, Inc. Terms and Conditions