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...
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2021-06-01
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| author | Gerlof P. Bosman Sergio Oliveira Peter J. Simons Javier Sastre Torano Govert W. Somsen Leon M. J. Knippels Rob Haselberg Roland J. Pieters Johan Garssen Karen Knipping |
| author_facet | Gerlof P. Bosman Sergio Oliveira Peter J. Simons Javier Sastre Torano Govert W. Somsen Leon M. J. Knippels Rob Haselberg Roland J. Pieters Johan Garssen Karen Knipping |
| author_sort | Gerlof P. Bosman |
| collection | DOAJ |
| 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 (<i>p</i> = 0.0001) as well as degranulation of anti-BLG IgE-sensitized RBL-hεIa-2B12 cells (<i>p</i> < 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. |
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| language | English |
| last_indexed | 2024-03-10T10:24:10Z |
| publishDate | 2021-06-01 |
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| spelling | doaj.art-31b0977593ed45d4b29fb83ded87455e2023-11-22T00:07:35ZengMDPI AGNutrients2072-66432021-06-01136204110.3390/nu13062041Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast CellsGerlof P. Bosman0Sergio Oliveira1Peter J. Simons2Javier Sastre Torano3Govert W. Somsen4Leon M. J. Knippels5Rob Haselberg6Roland J. Pieters7Johan Garssen8Karen Knipping9Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Universiteitsweg 99, 3584 CG Utrecht, The NetherlandsDanone Nutricia Research, Uppsalalaan 12, 3584 CT Utrecht, The NetherlandsPolpharma Biologics BV, Yalelaan 46, 3584 CM Utrecht, The NetherlandsDepartment of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Universiteitsweg 99, 3584 CG Utrecht, The NetherlandsDivision of Bioanalytical Chemistry, Amsterdam Institute for Molecules, Medicines and Life Sciences, Vrije Universiteit Amsterdam, de Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsDanone Nutricia Research, Uppsalalaan 12, 3584 CT Utrecht, The NetherlandsDivision of Bioanalytical Chemistry, Amsterdam Institute for Molecules, Medicines and Life Sciences, Vrije Universiteit Amsterdam, de Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsDepartment of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Universiteitsweg 99, 3584 CG Utrecht, The NetherlandsDanone Nutricia Research, Uppsalalaan 12, 3584 CT Utrecht, The NetherlandsDanone Nutricia Research, Uppsalalaan 12, 3584 CT Utrecht, The NetherlandsBackground: 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 (<i>p</i> = 0.0001) as well as degranulation of anti-BLG IgE-sensitized RBL-hεIa-2B12 cells (<i>p</i> < 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.https://www.mdpi.com/2072-6643/13/6/2041beta-lactoglobulinMaillard reactionlactosylationcow’s milk allergyantigenicitymast cell degranulation |
| spellingShingle | Gerlof P. Bosman Sergio Oliveira Peter J. Simons Javier Sastre Torano Govert W. Somsen Leon M. J. Knippels Rob Haselberg Roland J. Pieters Johan Garssen Karen Knipping Limited Lactosylation of Beta-Lactoglobulin from Cow’s Milk Exerts Strong Influence on Antigenicity and Degranulation of Mast Cells Nutrients beta-lactoglobulin Maillard reaction lactosylation cow’s milk allergy antigenicity mast cell degranulation |
| 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 | beta-lactoglobulin Maillard reaction lactosylation cow’s milk allergy antigenicity mast cell degranulation |
| url | https://www.mdpi.com/2072-6643/13/6/2041 |
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