The Journal of Experimental Medicine
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J. Exp. Med., Volume 189, Number 9, May 3, 1999 1507-1512

Humoral and Cell-mediated Autoimmune Reactions to Human Acidic Ribosomal P2 Protein in Individuals Sensitized to Aspergillus fumigatus P2 Protein

By Christina Mayer,* Ulrich Appenzeller,* Heike Seelbach,Dagger Gernot Achatz,§ Hannes Oberkofler,§ Michael Breitenbach,§ Kurt Blaser,* and Reto Crameri*

From the * Swiss Institute of Allergy and Asthma Research (SIAF), CH-7270 Davos, Switzerland; Dagger  Hochgebirgsklinik, CH-7265 Davos-Wolfgang, Switzerland; and the § Institut für Genetik und Allgemeine Biologie, Universität Salzburg,  A-5020 Salzburg, Austria

A panel of cDNAs encoding allergenic proteins was isolated from an Aspergillus fumigatus cDNA library displayed on the surface of filamentous phage. Solid phase-immobilized serum immunoglobulin E (IgE) from A. fumigatus-allergic individuals was used to enrich phage displaying IgE-binding molecules. One of the cDNAs encoded a 11.1-kD protein that was identified as acidic ribosomal phosphoprotein type 2 (P2 protein). The allergen, formally termed rAsp f 8, shares >62% sequence identity and >84% sequence homology to corresponding eukaryotic P2 proteins, including human P2 protein. The sequences encoding human and fungal P2 protein were subcloned, expressed in Escherichia coli as His6-tagged fusion proteins, and purified by Ni2+-chelate affinity chromatography. Both recombinant P2 proteins were recognized by IgE antibodies from allergic individuals sensitized to the A. fumigatus P2 protein and elicited strong type 1-specific skin reactions in these individuals. Moreover, human and fungal P2 proteins induced proliferative responses in peripheral blood mononuclear cells of A. fumigatus- allergic subjects sensitized to the fungal P2 protein. These data provide strong evidence for in vitro and in vivo humoral and cell-mediated autoreactivity to human P2 protein in patients suffering from chronic A. fumigatus allergy.

Key words: phage display;  cDNA libraries;  IgE;  allergens;  autoimmunity


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