Human IgE-binding epitopes of the latex allergen Hev b 5☆,☆☆,★
Section snippets
Antigens
Latex proteins were isolated from nonammoniated latex (NAL) by centrifugation to remove the rubber particles and particulates. The middle aqueous layer (C serum) was then dialyzed and lyophilized to concentrate the protein.
The cDNA for Hev b 5 was ligated into the expression vector pMAL-c2 (New England BioLabs, Beverly, Mass), which encodes maltose-binding protein (MBP), as previously described.12 The pMAL-c2/Hev b 5 plasmid was inserted into competent XL1 Blue Escherichia coli , and cultures
Anti-Hev b 5 antibodies
Recombinant Hev b 5 was produced by using the pMAL-c2 expression vector, which expresses the recombinant protein with the bacterial MBP. Pooled sera from health care workers allergic to latex strongly recognized the rHev b5/MBP construct (Fig 1).
DISCUSSION
Natural rubber latex is used in the manufacture of many different medical products. The latex is harvested from the H brasiliensis tree and processed into a material suitable for use in manufacturing. Harvesting and processing of the latex alters the native proteins present in latex and may alter their antigenic or allergenic properties.16, 19 Ammoniation and processing of the latex results in degradation of the native proteins and the potential exposure of cryptic epitopes hidden in the native
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Supported in part by a grant from Regent Medical, Norcross, Ga.
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Reprint requests: Donald Beezhold, PhD, Director, Laboratory of Immunobiology, Guthrie Research Institute, 1 Guthrie Square, Sayre, PA 18840.
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