IgE from latex-allergic patients binds to cloned and expressed B cell epitopes of prohevein.
Prohevein is one of the major allergens associated with latex allergy. In the present study, we identified IgE binding regions of prohevein, and expressed multiple IgE binding epitopes by selective cloning. These truncated polypeptides were then used to demonstrate IgE in the sera of patients. Decapeptides of prohevein were synthesized on derivatized cellulose membrane with an offset of one amino acid. The IgE reactivity of these linear peptides was evaluated separately using pooled sera from latex-allergic health care workers (HCW) and spina bifida (SB) patients. A total of 10 IgE binding epitopes representing unique as well as shared epitopes from both the N- and C-domains of the prohevein were identified. Recombinant polypeptides were constructed based on the identified epitopes, and clones carrying DNA fragments were overexpressed. These recombinant peptides were evaluated for IgE binding with sera from HCW, SB, and normal individuals. Recombinant prohevein, hevein, and the C-domain exhibited IgE binding in 84, 88, and 40% of HCW sera, respectively, as against reactivity of 84% with crude latex allergens. However, only 48% of the sera from SB patients showed IgE binding with recombinant prohevein, while 56 and 28% had reactivity with recombinant N- and C-domains, respectively. Among the three remaining recombinant peptides of the C-domain, only CA44-103 showed IgE binding with SB patients. The results of the present study suggest that linear IgE epitope analysis and construction of recombinant peptides increase the sensitivity and specificity of the immunodiagnosis of latex allergy and provide more information on the immunopathogenesis of hypersensitivity reaction mediated by type I allergy.
Banerjee
B
B
Allergy-Immunology Division of the Medical College of Wisconsin, Milwaukee 53226, USA.
Wang
X
X
Kelly
K J
KJ
Fink
J N
JN
Sussman
G L
GL
Kurup
V P
VP
D000485:Allergens; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001402:B-Lymphocytes / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D005069:Evaluation Studies as Topic; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology*; D007073:Immunoglobulin E / Q000378:metabolism*; D007840:Latex / Q000009:adverse effects*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010940:Plant Proteins / Q000276:immunology*; D011498:Protein Precursors / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D012882:Skin Tests
D000485:Allergens; D052179:Antigens, Plant; D000939:Epitopes; C097982:Hev b 6.01 protein, Hevea brasiliensis; D007840:Latex; D010446:Peptide Fragments; D010940:Plant Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D007073:Immunoglobulin E