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Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells
Background: beta-lactoglobulin (BLG) is one of the major cow’s milk proteins and the most abundant allergen in whey. Heating is a common technologic treatment applied during milk transformational processes. Maillardation of BLG in the presence of reducing sugars and elevated temperatures may influen...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8232271/ https://www.ncbi.nlm.nih.gov/pubmed/34203636 http://dx.doi.org/10.3390/nu13062041 |
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author | Bosman, Gerlof P. Oliveira, Sergio Simons, Peter J. Sastre Torano, Javier Somsen, Govert W. Knippels, Leon M. J. Haselberg, Rob Pieters, Roland J. Garssen, Johan Knipping, Karen |
author_facet | Bosman, Gerlof P. Oliveira, Sergio Simons, Peter J. Sastre Torano, Javier Somsen, Govert W. Knippels, Leon M. J. Haselberg, Rob Pieters, Roland J. Garssen, Johan Knipping, Karen |
author_sort | Bosman, Gerlof P. |
collection | PubMed |
description | Background: beta-lactoglobulin (BLG) is one of the major cow’s milk proteins and the most abundant allergen in whey. Heating is a common technologic treatment applied during milk transformational processes. Maillardation of BLG in the presence of reducing sugars and elevated temperatures may influence its antigenicity and allergenicity. Primary objective: to analyze and identify lactosylation sites by capillary electrophoresis mass spectrometry (CE-MS). Secondary objective: to assess the effect of lactosylated BLG on antigenicity and degranulation of mast cells. Methods: BLG was lactosylated at pH 7, a water activity (aw) of 0.43, and a temperature of 65 °C using a molar ratio BLG:lactose of 1:1 by incubating for 0, 3, 8, 16 or 24 h. For the determination of the effect on antibody-binding capacity of lactosylated BLG, an ELISA was performed. For the assessment of degranulation of the cell-line RBL-hεIa-2B12 transfected with the human α-chain, Fcε receptor type 1 (FcεRI) was used. Results: BLG showed saturated lactosylation between 8 and 16 incubation hours in our experimental setup. Initial stage lactosylation sites L1 (N-terminus)—K47, K60, K75, K77, K91, K138 and K141—have been identified using CE-MS. Lactosylated BLG showed a significant reduction of both the IgG binding (p = 0.0001) as well as degranulation of anti-BLG IgE-sensitized RBL-hεIa-2B12 cells (p < 0.0001). Conclusions and clinical relevance: this study shows that lactosylation of BLG decreases both the antigenicity and degranulation of mast cells and can therefore be a promising approach for reducing allergenicity of cow’s milk allergens provided that the process is well-controlled. |
format | Online Article Text |
id | pubmed-8232271 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-82322712021-06-26 Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells Bosman, Gerlof P. Oliveira, Sergio Simons, Peter J. Sastre Torano, Javier Somsen, Govert W. Knippels, Leon M. J. Haselberg, Rob Pieters, Roland J. Garssen, Johan Knipping, Karen Nutrients Article Background: beta-lactoglobulin (BLG) is one of the major cow’s milk proteins and the most abundant allergen in whey. Heating is a common technologic treatment applied during milk transformational processes. Maillardation of BLG in the presence of reducing sugars and elevated temperatures may influence its antigenicity and allergenicity. Primary objective: to analyze and identify lactosylation sites by capillary electrophoresis mass spectrometry (CE-MS). Secondary objective: to assess the effect of lactosylated BLG on antigenicity and degranulation of mast cells. Methods: BLG was lactosylated at pH 7, a water activity (aw) of 0.43, and a temperature of 65 °C using a molar ratio BLG:lactose of 1:1 by incubating for 0, 3, 8, 16 or 24 h. For the determination of the effect on antibody-binding capacity of lactosylated BLG, an ELISA was performed. For the assessment of degranulation of the cell-line RBL-hεIa-2B12 transfected with the human α-chain, Fcε receptor type 1 (FcεRI) was used. Results: BLG showed saturated lactosylation between 8 and 16 incubation hours in our experimental setup. Initial stage lactosylation sites L1 (N-terminus)—K47, K60, K75, K77, K91, K138 and K141—have been identified using CE-MS. Lactosylated BLG showed a significant reduction of both the IgG binding (p = 0.0001) as well as degranulation of anti-BLG IgE-sensitized RBL-hεIa-2B12 cells (p < 0.0001). Conclusions and clinical relevance: this study shows that lactosylation of BLG decreases both the antigenicity and degranulation of mast cells and can therefore be a promising approach for reducing allergenicity of cow’s milk allergens provided that the process is well-controlled. MDPI 2021-06-15 /pmc/articles/PMC8232271/ /pubmed/34203636 http://dx.doi.org/10.3390/nu13062041 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bosman, Gerlof P. Oliveira, Sergio Simons, Peter J. Sastre Torano, Javier Somsen, Govert W. Knippels, Leon M. J. Haselberg, Rob Pieters, Roland J. Garssen, Johan Knipping, Karen Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title | Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title_full | Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title_fullStr | Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title_full_unstemmed | Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title_short | Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells |
title_sort | limited lactosylation of beta-lactoglobulin from cow’s milk exerts strong influence on antigenicity and degranulation of mast cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8232271/ https://www.ncbi.nlm.nih.gov/pubmed/34203636 http://dx.doi.org/10.3390/nu13062041 |
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