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The Journal of Immunology, Vol 156, Issue 4 1618-1625, Copyright © 1996 by American Association of Immunologists


ARTICLES

The main IgE-binding epitope of a major latex allergen, prohevein, is present in its N-terminal 43-amino acid fragment, hevein

H Alenius, N Kalkkinen, T Reunala, K Turjanmaa and T Palosuo
National Public Health Institute, University of Helsinki, Finland.

Polypeptides of natural rubber latex (NRL) that elute from surgeon's gloves and other manufactured rubber products can sensitize exposed individuals and elicit severe hypersensitivity reactions. Previously, we showed that prohevein is a major allergen in NRL, the source material for rubber manufacturing. To analyze which region of the molecule carries the main IgE-binding epitope(s), we purified prohevein and its C-domain from NRL by gel filtration, reverse phase chromatography, and electroelution. In immunoblotting, prohevein bound IgE from 15 of 20 (75%), and the prohevein C-domain from 3 of 20 (15%) latex-allergic patient sera. In ELISA, 36 of 52 (69%) patient sera showed IgE binding to prohevein, whereas 11 of 52 (21%) sera had IgE to prohevein C-domain. We then purified from a brand of highly allergenic surgeon's gloves six hydrophilic peptides that revealed in amino- terminal sequencing 100% identity to the N-terminus of prohevein. In mass spectrometry, all purified peptides gave a molecular mass of 4719 +/- 1.9 daltons, which corresponds to the molecular mass of hevein (4719.1 daltons), a 43-amino acid N-terminal fragment of prohevein. Purified hevein inhibited 72% of IgE binding from pooled sera of NRL- allergic patients to solid phase glove extract and 45% of IgE binding to solid phase NRL. Of the 43 NRL-allergic patient sera tested, 56% showed IgE Abs to purified hevein in ELISA. In skin prick testing, purified hevein elicited positive reactions in three-quarters of the latex-allergic patients tested. These results indicate that the majority of prohevein's IgE-binding ability resides in its N-terminal fragment, known as hevein. In one highly allergenic latex glove examined, the majority of IgE-binding ability was attributable to hevein molecules, suggesting that these peptides can be significant sensitizers in NRL allergy.


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