Allergy Asthma Immunol Res.  2018 Jul;10(4):363-369. 10.4168/aair.2018.10.4.363.

Immunoglobulin E-binding Proteins of Cooked Walnuts in Korean Children

Affiliations
  • 1Department of Pediatrics, Ajou University School of Medicine, Suwon, Korea. jsjs87@ajou.ac.kr

Abstract

PURPOSE
The immunological characteristics of young Korean children with walnut (WN) allergy and the influence of different cooking methods on WN proteins have not been evaluated to date. This study aimed to evaluate the major WN allergens identified among Korean children, together with changes in WN antigenicity caused by common cooking methods.
METHODS
We enrolled children under the age of 13 years with WN serum-specific immunoglobulin (Ig) E concentrations. The protein fractions of dry-fried and boiled WN extracts were compared with those of raw WNs using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), 2-dimentional gel electrophoresis (2DE) and a proteomic analysis using electrospray ionization (liquid chromatography-mass spectrometry [LC-MS]). An immunoblotting analysis was conducted to examine IgE reactivity toward raw WNs using serum samples from 6 children with a clinical WN allergy. To determine the processed WN proteins with IgE-binding capacity, a 2D-immunoblotting analysis was performed using the pooled sera of 20 WN-sensitized children.
RESULTS
Protein bands from raw WNs were identified at 9, 16, 28, 52, 58, and 64 kDa via SDS-PAGE. The 9- and 16-kDa protein bands were enhanced by boiling, whereas the 52- and 64-kDa bands were considerably diminished. On LC-MS analysis, of the 66 IgE-binding proteins present in raw WNs, 57 were found in dry-fried WNs, but only 4 in boiled WNs. The sera of 5 out of 6 participants reacted with the 52-kDa protein bands and those of 4 out of 6 participants reacted with the 16- and 28-kDa protein bands, respectively. Meanwhile, a 2D-immunoblotting result confirmed the presence of different binding patterns among children who consumed cooked WNs.
CONCLUSIONS
The protein profile of boiled WNs is substantially different from that of raw WNs. However, 4 proteins including prolamins remained stable after dry-frying or boiling. Further studies are needed to evaluate the clinical relevance of these findings.

Keyword

Antigenicity; cooking; walnut

MeSH Terms

Allergens
Child*
Cooking
Electrophoresis
Electrophoresis, Polyacrylamide Gel
Humans
Hypersensitivity
Immunoblotting
Immunoglobulin E
Immunoglobulins*
Juglans*
Prolamins
Sodium Dodecyl Sulfate
Spectrum Analysis
Allergens
Immunoglobulin E
Immunoglobulins
Prolamins
Sodium Dodecyl Sulfate

Figure

  • Fig. 1 Overview of the methodology used in this study to characterize the proteome of processed walnut.

  • Fig. 2 SDS-PAGE analysis of processed walnut extract (M, marker; 1, raw; 2, dry-frying; 3, boiling). Concentration of extracts: raw 4.4 mg/mL; dry-frying 3.4 mg/mL; boiling 17.0 mg/mL. IgE bands identified: 9, 16, 28, 52, 58, 64 kDa.

  • Fig. 3 2DE analysis of processed walnut extract (1, raw; 2, dry-frying; 3, boiling; M.W, molecular weight).

  • Fig. 4 Immunoblot analysis of raw walnut extract using the sera of patients with clinical walnut allergy (MW, molecular weight; B, BSA; N, negative control; P, patient).

  • Fig. 5 2D-Immunoblot analysis of processed walnut extract using walnut specific IgE positive pooled sera of patients (1, raw; 2, dry-frying; 3, boiling; M.W, molecular weight).

  • Fig. 6 Overview of the number of protein bands observed in processed walnut extracts via the proteomic approach. Out of the 66 proteins present in raw WNs, only 4 were observed in boiled WNs. GI, GenInfo; NCBI, National Center for Biotechnology Information non-redundant database.


Cited by  1 articles

Is There Any Necessity to Prescribe Consumption of Walnuts Cooked by Different Processing Techniques to Patients With Walnut Allergy?
Jae-Won Oh
Allergy Asthma Immunol Res. 2018;10(4):287-289.    doi: 10.4168/aair.2018.10.4.287.


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