α-Gal on the protein surface affects uptake and degradation in immature monocyte derived dendritic cells

Scientific Reports
M Krstić RistivojevićM van Hage

Abstract

Red meat allergy is characterized by an IgE response against the carbohydrate galactose-α-1,3-galactose (α-Gal), which is abundantly expressed on glycoproteins from non-primate mammals. The mechanisms of how α-Gal is processed and presented to the immune system to initiate an allergic reaction are still unknown. The aim of this study was to reveal whether the presence of α-Gal epitopes on the protein surface influence antigen uptake and processing in immature monocyte-derived dendritic cells (iMDDCs). Immature MDDCs were prepared from healthy blood donors and red meat allergic patients. We found an increased internalization of α-Gal carrying proteins over time in iMDDCs by flow cytometric analysis, which was independent of the donor allergic status. The uptake of α-Gal carrying proteins was significantly higher than the uptake of non-α-Gal carrying proteins. Confocal microscopy revealed α-Gal carrying proteins scattered around the cytoplasm in most iMDDCs while detection of proteins not carrying α-Gal was negligible. Fluorescent detection of protein on SDS-PAGE showed that degradation of α-Gal carrying proteins was slower than degradation of non-α-Gal carrying proteins. Thus, the presence of α-Gal on the protein surface affects...Continue Reading

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Citations

Jun 17, 2020·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·Krzyszof RutkowskiAnna Moniuszko-Malinowska
Jun 20, 2019·Frontiers in Immunology·Alejandro Cabezas-CruzJosé de la Fuente
Jun 7, 2019·F1000Research·Andrea J LukerRebecca K Martin
Nov 10, 2020·Frontiers in Immunology·Miriam HilsTilo Biedermann

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Methods Mentioned

BETA
glycosylation
protein assay
density gradient centrifugation
Scanning Microscopy

Software Mentioned

BD FACS Diva
GraphPad Prism
FlowJo
GraphPad
Zeiss LSM Image Browser

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