Projekte

Only alpha-Gal bound to lipids, but not to proteins, is transported across enterocytes as an IgE-reactive molecule that can induce effector cell activation

Autor(en)
Patricia Roman-Carrasco, Barbara Lieder, Veronika Somoza, Marta Ponce, Zsolt Szepfalusi, Diana Martin, Wolfgang Hemmer, Ines Swoboda
Abstrakt

Background The oligosaccharide galactose-alpha-1,3-galactose (alpha-Gal), present in mammalian proteins and lipids, causes an unusual delayed allergic reaction 3 to 6 hours after ingestion of mammalian meat in individuals with IgE antibodies against alpha-Gal. To better understand the delayed onset of allergic symptoms and investigate whether protein-bound or lipid-bound alpha-Gal causes these symptoms, we analyzed the capacity of alpha-Gal conjugated proteins and lipids to cross a monolayer of intestinal cells. Methods Extracts of proteins and lipids from beef were prepared, subjected to in vitro digestions, and added to Caco-2 cells grown on permeable supports. The presence of alpha-Gal in the basolateral medium was investigated by immunoblotting, thin-layer chromatography with immunostaining and ELISA, and its allergenic activity was analyzed in a basophil activation test. Results After addition of beef proteins to the apical side of Caco-2 cells, alpha-Gal containing peptides were not detected in the basolateral medium. Those peptides that crossed the Caco-2 monolayer did not activate basophils from an alpha-Gal allergic patient. Instead, when Caco-2 cells were incubated with lipids extracted from beef, alpha-Gal was detected in the basolateral medium. Furthermore, these alpha-Gal lipids were able to activate the basophils of an alpha-Gal allergic patient in a dose-dependent manner. Conclusion Only alpha-Gal bound to lipids, but not to proteins, is able to cross the intestinal monolayer and trigger an allergic reaction. This suggests that the slower digestion and absorption of lipids might be responsible for the unusual delayed allergic reactions in alpha-Gal allergic patients and identifies glycolipids as potential allergenic molecules.

Organisation(en)
Institut für Physiologische Chemie, Department für Mikrobiologie und Ökosystemforschung
Externe Organisation(en)
FH Campus Wien, Medizinische Universität Wien, CSIC UAM, Consejo Superior de Investigaciones Cientificas (CSIC), CSIC-UAM - Instituto de Investigacion en Ciencias de la Alimentacion (CIAL), Inst Invest Ciencias Alimentac CIAL, Dept Prod & Caracterizac Nuevos Alimentos, FAZ Floridsdorf Allergy Ctr
Journal
Allergy: European journal of allergy and immunology
Band
74
Seiten
1956-1968
ISSN
0105-4538
DOI
https://doi.org/10.1111/all.13873
Publikationsdatum
10-2019
Peer-reviewed
Ja
ÖFOS 2012
Allergologie, Immunologie
Schlagwörter
Link zum Portal
https://ucris.univie.ac.at/portal/de/publications/only-alphagal-bound-to-lipids-but-not-to-proteins-is-transported-across-enterocytes-as-an-igereactive-molecule-that-can-induce-effector-cell-activation(86ba8bef-6f93-413a-b6a9-965278222ad5).html

Publikationen

Only alpha-Gal bound to lipids, but not to proteins, is transported across enterocytes as an IgE-reactive molecule that can induce effector cell activation

Autor(en)
Patricia Roman-Carrasco, Barbara Lieder, Veronika Somoza, Marta Ponce, Zsolt Szepfalusi, Diana Martin, Wolfgang Hemmer, Ines Swoboda
Abstrakt

Background The oligosaccharide galactose-alpha-1,3-galactose (alpha-Gal), present in mammalian proteins and lipids, causes an unusual delayed allergic reaction 3 to 6 hours after ingestion of mammalian meat in individuals with IgE antibodies against alpha-Gal. To better understand the delayed onset of allergic symptoms and investigate whether protein-bound or lipid-bound alpha-Gal causes these symptoms, we analyzed the capacity of alpha-Gal conjugated proteins and lipids to cross a monolayer of intestinal cells. Methods Extracts of proteins and lipids from beef were prepared, subjected to in vitro digestions, and added to Caco-2 cells grown on permeable supports. The presence of alpha-Gal in the basolateral medium was investigated by immunoblotting, thin-layer chromatography with immunostaining and ELISA, and its allergenic activity was analyzed in a basophil activation test. Results After addition of beef proteins to the apical side of Caco-2 cells, alpha-Gal containing peptides were not detected in the basolateral medium. Those peptides that crossed the Caco-2 monolayer did not activate basophils from an alpha-Gal allergic patient. Instead, when Caco-2 cells were incubated with lipids extracted from beef, alpha-Gal was detected in the basolateral medium. Furthermore, these alpha-Gal lipids were able to activate the basophils of an alpha-Gal allergic patient in a dose-dependent manner. Conclusion Only alpha-Gal bound to lipids, but not to proteins, is able to cross the intestinal monolayer and trigger an allergic reaction. This suggests that the slower digestion and absorption of lipids might be responsible for the unusual delayed allergic reactions in alpha-Gal allergic patients and identifies glycolipids as potential allergenic molecules.

Organisation(en)
Institut für Physiologische Chemie, Department für Mikrobiologie und Ökosystemforschung
Externe Organisation(en)
FH Campus Wien, Medizinische Universität Wien, CSIC UAM, Consejo Superior de Investigaciones Cientificas (CSIC), CSIC-UAM - Instituto de Investigacion en Ciencias de la Alimentacion (CIAL), Inst Invest Ciencias Alimentac CIAL, Dept Prod & Caracterizac Nuevos Alimentos, FAZ Floridsdorf Allergy Ctr
Journal
Allergy: European journal of allergy and immunology
Band
74
Seiten
1956-1968
ISSN
0105-4538
DOI
https://doi.org/10.1111/all.13873
Publikationsdatum
10-2019
Peer-reviewed
Ja
ÖFOS 2012
Allergologie, Immunologie
Schlagwörter
Link zum Portal
https://ucris.univie.ac.at/portal/de/publications/only-alphagal-bound-to-lipids-but-not-to-proteins-is-transported-across-enterocytes-as-an-igereactive-molecule-that-can-induce-effector-cell-activation(86ba8bef-6f93-413a-b6a9-965278222ad5).html

Vortraege

Only alpha-Gal bound to lipids, but not to proteins, is transported across enterocytes as an IgE-reactive molecule that can induce effector cell activation

Autor(en)
Patricia Roman-Carrasco, Barbara Lieder, Veronika Somoza, Marta Ponce, Zsolt Szepfalusi, Diana Martin, Wolfgang Hemmer, Ines Swoboda
Abstrakt

Background The oligosaccharide galactose-alpha-1,3-galactose (alpha-Gal), present in mammalian proteins and lipids, causes an unusual delayed allergic reaction 3 to 6 hours after ingestion of mammalian meat in individuals with IgE antibodies against alpha-Gal. To better understand the delayed onset of allergic symptoms and investigate whether protein-bound or lipid-bound alpha-Gal causes these symptoms, we analyzed the capacity of alpha-Gal conjugated proteins and lipids to cross a monolayer of intestinal cells. Methods Extracts of proteins and lipids from beef were prepared, subjected to in vitro digestions, and added to Caco-2 cells grown on permeable supports. The presence of alpha-Gal in the basolateral medium was investigated by immunoblotting, thin-layer chromatography with immunostaining and ELISA, and its allergenic activity was analyzed in a basophil activation test. Results After addition of beef proteins to the apical side of Caco-2 cells, alpha-Gal containing peptides were not detected in the basolateral medium. Those peptides that crossed the Caco-2 monolayer did not activate basophils from an alpha-Gal allergic patient. Instead, when Caco-2 cells were incubated with lipids extracted from beef, alpha-Gal was detected in the basolateral medium. Furthermore, these alpha-Gal lipids were able to activate the basophils of an alpha-Gal allergic patient in a dose-dependent manner. Conclusion Only alpha-Gal bound to lipids, but not to proteins, is able to cross the intestinal monolayer and trigger an allergic reaction. This suggests that the slower digestion and absorption of lipids might be responsible for the unusual delayed allergic reactions in alpha-Gal allergic patients and identifies glycolipids as potential allergenic molecules.

Organisation(en)
Institut für Physiologische Chemie, Department für Mikrobiologie und Ökosystemforschung
Externe Organisation(en)
FH Campus Wien, Medizinische Universität Wien, CSIC UAM, Consejo Superior de Investigaciones Cientificas (CSIC), CSIC-UAM - Instituto de Investigacion en Ciencias de la Alimentacion (CIAL), Inst Invest Ciencias Alimentac CIAL, Dept Prod & Caracterizac Nuevos Alimentos, FAZ Floridsdorf Allergy Ctr
Journal
Allergy: European journal of allergy and immunology
Band
74
Seiten
1956-1968
ISSN
0105-4538
DOI
https://doi.org/10.1111/all.13873
Publikationsdatum
10-2019
Peer-reviewed
Ja
ÖFOS 2012
Allergologie, Immunologie
Schlagwörter
Link zum Portal
https://ucris.univie.ac.at/portal/de/publications/only-alphagal-bound-to-lipids-but-not-to-proteins-is-transported-across-enterocytes-as-an-igereactive-molecule-that-can-induce-effector-cell-activation(86ba8bef-6f93-413a-b6a9-965278222ad5).html