Dataset Information

Preparation, Identification and Application of β-Lactoglobulin Hydrolysates with Oral Immune Tolerance.

ABSTRACT: To reveal, for the first time, the mechanism of T cell epitope release from β-lactoglobulin that induces oral immune tolerance, a strategy for the prediction, preparation, identification and application of β-lactoglobulin hydrolysate with oral immune tolerance was established using the bioinformatics method, hydrolysis, mass spectrometry, T cell proliferation assays and animal experiments. Some T cell epitope peptides of β-lactoglobulin were identified for the first time. The hydrolysates of trypsin, protamex and papain showed oral tolerance, among which the hydrolysates of protamex and papain have been reported for the first time. Although the neutral protease hydrolysate contained T cell epitopes, it still had allergenicity. The mechanism behind oral immune tolerance induction by T cell epitopes needs to be further revealed. In addition, the trypsin hydrolysate with abundant T cell epitopes was added to whey protein to prepare the product for oral immune tolerance. Overall, this study provides insights into the development of new anti-allergic milk-based products and their application in the clinical treatment of milk allergies.

SUBMITTER: Tian L

PROVIDER: S-EPMC9857568 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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Preparation, Identification and Application of β-Lactoglobulin Hydrolysates with Oral Immune Tolerance.

Tian Linghan L Zhang Qianqian Q Cong Yanjun Y Yan Wenjie W

Foods (Basel, Switzerland) 20230109 2

To reveal, for the first time, the mechanism of T cell epitope release from β-lactoglobulin that induces oral immune tolerance, a strategy for the prediction, preparation, identification and application of β-lactoglobulin hydrolysate with oral immune tolerance was established using the bioinformatics method, hydrolysis, mass spectrometry, T cell proliferation assays and animal experiments. Some T cell epitope peptides of β-lactoglobulin were identified for the first time. The hydrolysates of try ...[more]

PMID: 36673400

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Project description:BackgroundCow's milk-derived whey hydrolysates are nutritional substitutes for allergic infants. Safety or residual allergenicity assessment of these whey hydrolysates is crucial. Currently, rat basophilic leukemia RBL-2H3 cells expressing the human IgE receptor α-chain (huFcεRIα-RBL-2H3), sensitized with serum IgE from cow's milk allergic children, are being employed to assess in vitro residual allergenicity of these whey hydrolysates. However, limited availability and inter-lot variation of these allergic sera impede standardization of whey hydrolysate safety testing in degranulation assays.ObjectiveAn oligoclonal pool of chimeric human (chu)IgE antibodies against bovine β-lactoglobulin (a major allergen in whey) was generated to increase sensitivity, specificity, and reproducibility of existing degranulation assays.MethodsMice were immunized with bovine β-lactoglobulin, and subsequently the variable domains of dissimilar anti-β-lactoglobulin mouse IgG antibodies were cloned and sequenced. Six chimeric antibodies were generated comprising mouse variable domains and human constant IgE/κ domains.ResultsAfter sensitization with this pool of anti-β-lactoglobulin chuIgEs, huFcεRIα-expressing RBL-2H3 cells demonstrated degranulation upon cross-linking with whey, native 18 kDa β-lactoglobulin, and 5-10 kDa whey hydrolysates, whereas a 3 kDa whey hydrolysate and cow's milk powder (mainly casein) showed no degranulation. In parallel, allergic serum IgEs were less sensitive. In addition, our pool anti-β-lactoglobulin chuIgEs recognized multiple allergenic immunodominant regions on β-lactoglobulin, which were also recognized by serum IgEs from cow's milk allergic children.ConclusionUsage of our 'unlimited' source and well-defined pool of β-lactoglobulin-specific recombinant chuIgEs to sensitize huFcεRIα on RBL-2H3 cells showed to be a relevant and sensitive alternative for serum IgEs from cow's milk allergic patients to assess safety of whey-based non-allergic hydrolyzed formula.

Dataset Information

Preparation, Identification and Application of β-Lactoglobulin Hydrolysates with Oral Immune Tolerance.

Publications

Preparation, Identification and Application of β-Lactoglobulin Hydrolysates with Oral Immune Tolerance.

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