Assessment of the sensitizing capacity and allergenicity of enzymatic cross-linked arginine kinase, the crab allergen

• Published in: Molecular nutrition & food research 2016-07, Vol.60 (7), p.1707-1718
• Main Authors: Fei, Dan-Xia, Liu, Qing-Mei, Chen, Feng, Yang, Yang, Chen, Zhong-Wei, Cao, Min-Jie, Liu, Guang-Ming
• Format: Article
• Language: English
• Subjects: Adult; Allergenicity; Allergens - analysis; Allergens - immunology; Animals; Arginine kinase; Arginine Kinase - immunology; Arginine Kinase - metabolism; Brachyura - immunology; Cambaridae; Cell Line, Tumor; Decapoda; Digestion; Disease Models, Animal; Enzymatic cross-linking; Female; Humans; Immunoglobulin E - immunology; Infant; Injections, Intraperitoneal; Male; Mice; Mice, Inbred BALB C; Middle Aged; Rats; Risk Factors; Shellfish; Shellfish Hypersensitivity - immunology; Shellfish Hypersensitivity - prevention & control; Thermal polymerization; Tyrosinase; Young Adult
• ISSN: 1613-4125; 1613-4133
• DOI: 10.1002/mnfr.201500936

Scope The enzymatic cross‐linking of an allergen by food processing may alter its sensitization potential. In this study, the IgE‐binding activity and allergenicity of cross‐linked thermal polymerized arginine kinase (CL‐pAK) were investigated. Methods and results The IgE‐binding activity and stability of CL‐pAK were analyzed by immunological and proteomics methods. The sensitization and potency to induce oral tolerance of CL‐pAK were tested using in vivo assays and a cell model. According to the results of inhibition of ELISA, the half inhibitory concentration of AK after cross‐linking changed from 1.13 to 228.36 μg/mL. The results of in vitro digestion demonstrated that CL‐pAK showed more resistance to gastrointestinal digestion than native AK. Low allergenicity and capacity to induce oral tolerance in mice were shown by the sera levels of AK‐specific antibodies and T‐cell cytokine production. Exposure of RBL‐2H3 cells to CL‐pAK compared with AK, resulted in lower levels of mast degranulation and histamine. Conclusion Enzymatic cross‐linking with thermal polymerization of AK by tyrosinase and caffeic acid had high potential in mitigating IgE‐binding activity and allergenicity, which were influenced by altering the molecular and immunological features of the shellfish protein. Arginine kinase (AK) is a major allergen of crab, shrimp, and crayfish, and is likely to be found in allergic and cross‐activity reactions in different species. In this study, the polymerized AK is effectively cross‐linked by the tyrosinase and caffeic acid. The resulting CL‐pAK has high potential in mitigating IgE‐binding activity and a low sensitization capacity to CL‐pAK. We infer that cross‐linking could reduce the IgE‐binding activity of AK due to its antigen epitopes covered. This suggests that CL‐pAK may reduce the risk of shellfish allergy in humans, although clinical trials are required for validation.