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- Title
Recombinant high molecular weight-glutenin subunit-specific IgE detection is useful in identifying wheat-dependent exercise-induced anaphylaxis complementary to recombinant omega-5 gliadin-specific IgE test.
- Authors
Takahashi, H.; Matsuo, H.; Chinuki, Y.; Kohno, K.; Tanaka, A.; Maruyama, N.; Morita, E.
- Abstract
Background Wheat-dependent exercise-induced anaphylaxis ( WDEIA) is a special form of food allergy typically induced by exercise after ingestion of wheat products. We identified wheat omega-5 gliadin and high molecular weight-glutenin subunit ( HMW-glutenin) as major allergens for WDEIA and clarified that simultaneous detection of serum IgE binding to synthetic epitope peptides of these allergens identifies more than 90% of WDEIA patients. However, the short synthetic peptides are not suitable for CAP-fluorescent enzyme-immunoassay ( CAP- FEIA), which is widely utilized for detecting allergen-specific IgE. Objective In this study, we constructed a CAP- FEIA with recombinant HMW-glutenin, and evaluated its usefulness in identifying the patients with WDEIA. Methods Recombinant HMW-glutenin was expressed as histidine-tag protein in E. coli and purified by histidine-tag affinity column. Wheat, gluten, recombinant omega-5 gliadin, epitope peptide of HMW-glutenin, native and recombinant HMW-glutenin specific IgE in the sera from 48 patients with WDEIA, 16 patients with atopic dermatitis ( AD) who had no immediate allergic reaction after wheat ingestion and 12 healthy controls were determined by using CAP- FEIA method. Results In 16 AD patients without wheat allergy 12 of them (75%) had positive results for native HMW-glutenin test in contrast to epitope peptide of HMW-glutenin (12.5%) and recombinant HMW-glutenin test (12.5%). These results indicate the native HMW-glutenin test has low specificity. Sensitivity and specificity of the IgE test with recombinant HMW-glutenin were 16.7% and 92.9%. These are well compatible with results obtained by using epitope peptide of HMW-glutenin. However, sensitivity and specificity reached to 93.8% and 92.9%, when the test was combined to the test with recombinant omega-5 gliadin. Conclusions and Clinical Relevance We demonstrated that recombinant HMW-glutenin is best for CAP- FEIA system in point of stability and specificity and confirmed that detection of specific IgE against recombinant HMW-glutenin is useful for diagnosis of WDEIA when combined with the CAP- FEIA (recombinant omega-5 gliadin) test.
- Subjects
ANAPHYLAXIS; ALLERGIES; FOOD allergy; GLUTELINS; GLIADINS; IMMUNOGLOBULIN E; IMMUNOGLOBULINS
- Publication
Clinical & Experimental Allergy, 2012, Vol 42, Issue 8, p1293
- ISSN
0954-7894
- Publication type
Article
- DOI
10.1111/j.1365-2222.2012.04039.x