Original Articles: Environmental and Occupational DisordersMutational analysis of the IgE epitopes in the latex allergen Hev b 5☆,☆☆
Section snippets
Antibodies
Human sera with anti-Hev b 5 IgE were obtained from 7 health-care workers diagnosed with latex allergy who had positive skin test responses to recombinant (r) Hev b 5 and demonstrated skin test reactivity at 1 ng/mL or less.15 Rabbit anti-Hev b 5 IgG was obtained from a New Zealand white rabbit immunized with 200 μg of maltose-binding protein/rHev b 5 in a 50:50 mixture of TiterMax adjuvant (CytRx, Norcross, Ga). The rabbit was given a booster injection at 3 months and was bled 4 weeks later,
Immunodominant Hev b 5 epitopes
We have previously reported 6 epitopes for human IgE (epitopes 1, 2, 4, 6, 7, and 10) that were identified by using SPOTs analysis with sera pooled from 10 health-care workers with latex allergy.17 An additional IgE-binding epitope (epitope 8) was detected by using sera from an individual patient with latex allergy (unpublished data). We now examine the Hev b 5 sequence by using sera from a second group of 7 patients selected because they had a positive skin prick test response to 1 ng/mL or
Discussion
Patients with latex allergy are often hyperreactive to latex protein, making immunotherapy risky. We have shown that recombinant latex proteins are an important source of purified latex allergens that may serve to improve in vitro and in vivo diagnostic tests.15 The production of recombinant proteins and the potential to modify the cDNA for these proteins allow novel approaches for immunotherapy. It has been proposed that site-directed mutagenesis be used to produce hypoallergenic variants of
Acknowledgements
We thank Maren Nalepa, Colleen Landy, Ben McClintic, and Deidre Blake for excellent technical assistance.
References (23)
- et al.
Hypersensitivity to natural latex
J Allergy Clin Immunol
(1989) - et al.
Latex-associated allergies and anaphylactic reactions
Clin Immunol Immunopathol
(1992) - et al.
Cornstarch powder on latex products is an allergen carrier
J Allergy Clin Immunol
(1994) - et al.
Specific immunotherapy with a standardized latex extract versus placebo in allergic healthcare workers
J Allergy Clin Immunol
(2000) - et al.
A novel acidic allergen, Hev b 5, in latex
J Biol Chem
(1996) - et al.
Identification, cloning, and sequence of a major allergen (Hev b 5) from natural rubber latex (Hevea brasiliensis)
J Biol Chem
(1996) - et al.
Murine B-cell and T-cell epitopes of the allergen Hev b 5 from natural rubber latex
Mol Immunol
(1999) - et al.
Human IgE-binding epitopes of the latex allergen Hev b 5
J Allergy Clin Immunol
(1999) - et al.
Human T-cell epitopes of the latex allergen Hev b 5 in health care workers
J Allergy Clin Immunol
(2000) - et al.
Recombinant allergens for diagnosis and therapy of allergic disease
J Allergy Clin Immunol
(2000)
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ENEA, a peach and apricot IgE-binding protein cross-reacting with the latex major allergen Hev b 5
2019, Molecular ImmunologyCitation Excerpt :However, this difference could be due to the characteristics of the nENEA sample used, that was indeed a mixture of fragments rather than the entire molecule, thus not displaying its overall natural structure. Both nENEA and rENEA were able to partially inhibit the IgE binding to Hev b 5 immobilized on the beads of the ABA system.This result is in line with the observation that the human IgE epitopes identified in Hev b 5 (Beezhold et al., 1999, 2001) are only partially conserved in ENEA. It is conceivable that the most conserved antigenic determinants may be involved in the observed cross-reactivity between ENEA and Hev b 5.
In silico locating the immune-reactive segments of Lepidium draba peroxidase and designing a less immune-reactive enzyme derivative
2017, Computational Biology and ChemistryCitation Excerpt :Intense effects of single amino acid change on the affinity of immunoglobulins to epitops have been reported previously (Chen et al., 2012). For example, The K274A mutation largely abolished IgE-binding ability of Hev b 5 which is an acidic protein and the major latex allergen (Beezhold et al., 2001). The phenomenon is attributed to charge and/or size differences between amino acids.
Mechanisms underlying allergy vaccination with recombinant hypoallergenic allergen derivatives
2012, VaccineCitation Excerpt :The strategies described above allow the production of well-defined molecules (regarding their biochemical and immunological characteristics as well as the production process) in a stable and reproducible way, which cannot be achieved by chemical modifications of the corresponding recombinant wildtype allergen [65,66]. Beside the loss of conformation and therefore IgE reactivity, amino acids, which are directly involved in the allergen–IgE interaction, were targeted by site-directed mutagenesis [67,68]. However, IgE epitopes are difficult to identify, as IgE directed to a conformational epitope normally does not bind to linear peptides [69].
3-D Model of the bee venom acid phosphatase: Insights into allergenicity
2009, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Two of them (res. 153–163 and 336–339) contain the sequence XEEX which is present also in six human IgE binding regions of the allergen Hev b 5 [28]. Moreover, such sequences are present in the rabbit IgG binding epitopes of the same allergen [33].
Hypoallergens for allergen-specific immunotherapy by directed molecular evolution of mite group 2 allergens
2007, Journal of Biological Chemistry
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Supported by the LEAP Testing Service of the Guthrie Research Institute.
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Reprint requests: Donald Beezhold, PhD, Director, Laboratory of Immunobiology, Guthrie Research Institute, 1 Guthrie Square, Sayre, PA 18840.