Supplementary data for article: Apostolovic, D.; Krstic, M.; Mihailovic, J.; Starkhammar, M.; Cirkovic Velickovic, T.; Hamsten, C.; Van Hage, M. Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-Reactive Peptides. Scientific Reports 2017, 7 (1). https://doi.org/10.1038/s41598-017-05355-4

Supplementary material for: [https://doi.org/10.1038/s41598-017-05355-4]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2486

Supplementary data for article: Apostolovic, D.; Tran, T. A. T.; Sánchez-Vidaurre, S.; Cirkovic Velickovic, T.; Starkhammar, M.; Hamsten, C.; Van Hage, M. Red Meat Allergic Patients Have a Selective IgE Response to the α-Gal Glycan. Allergy: European Journal of Allergy and Clinical Immunology 2015, 70 (11), 1497–1500. https://doi.org/10.1111/all.12695

Supporting information for: [https://doi.org/10.1111/all.12695]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1982

Supplementary data for the article: Ristivojević, M. K.; Grundström, J.; Tran, T. A. T.; Apostolovic, D.; Radoi, V.; Starkhammar, M.; Vukojević, V.; Ćirković Veličković, T.; Hamsten, C.; van Hage, M. α-Gal on the Protein Surface Affects Uptake and Degradation in Immature Monocyte Derived Dendritic Cells. Scientific Reports 2018, 8 (1). https://doi.org/10.1038/s41598-018-30887-8

Supplementary material for: [https://doi.org/10.1038/s41598-018-30887-8]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2207

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

Red meat allergy is characterized by an IgE response against the carbohydrate galactose-alpha-1,3-galactose (alpha-Gal), which is abundantly expressed on glycoproteins from non-primate mammals. The mechanisms of how alpha-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 alpha-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 alpha-Gal carrying proteins over time in iMDDCs by flow cytometric analysis, which was independent of the donor allergic status. The uptake of alpha-Gal carrying proteins was significantly higher than the uptake of non-alpha-Gal carrying proteins. Confocal microscopy revealed alpha-Gal carrying proteins scattered around the cytoplasm in most iMDDCs while detection of proteins not carrying alpha-Gal was negligible. Fluorescent detection of protein on SDS-PAGE showed that degradation of alpha-Gal carrying proteins was slower than degradation of non-alpha-Gal carrying proteins. Thus, the presence of alpha-Gal on the protein surface affects both uptake and degradation of the protein, and the results add new knowledge of alpha-Gal as a clinically relevant food allergen.Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3073

Supplementary data for the article: Apostolovic, D.; Sánchez-Vidaurre, S.; Waden, K.; Curin, M.; Grundström, J.; Gafvelin, G.; Cirkovic Velickovic, T.; Grönlund, H.; Thomas, W. R.; Valenta, R.; et al. The Cat Lipocalin Fel d 7 and Its Cross-Reactivity with the Dog Lipocalin Can f 1. Allergy: European Journal of Allergy and Clinical Immunology 2016, 71 (10), 1490–1495. https://doi.org/10.1111/all.12955

Supplementary material for: [https://doi.org/10.1111/all.12955]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2337]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3650

Cross-reactivity between tick and wasp venom can contribute to frequent wasp sensitization in patients with the α-Gal syndrome

Background α-Gal syndrome (AGS) is a food allergy with severe delayed allergic reactions, mediated by IgE-reactivity to galactose-α1,3-galactose (α-Gal). AGS is strongly associated with tick bites. An increased incidence of venom sensitization has been found in AGS patients. Here, we evaluated the frequency of wasp sensitization in Swedish AGS patients and the possible cross-reactivity between wasp venom and tick proteins. Methods Sera from 136 Swedish AGS patients and 29 wasp-positive non-AGS control sera were analyzed for IgE-reactivity against wasp venom (Vespula spp.), the European tick Ixodes ricinus (Streptavidin ImmunoCAP), α-Gal and total IgE by ImmunoCAP. The presence of α-Gal on wasp venom proteins (Vespula vulgaris) was investigated by western blot (WB), and possible cross-reactivity between wasp venom and tick proteins by enzyme-linked immunosorbent assay and WB. Involvement of cross-reactive carbohydrate domains (CCDs) was also assessed. Results Wasp sensitization was present in 54% of AGS patients, although the IgE levels were low. Wasp sensitized patients had higher IgE levels to α-Gal and total IgE levels compared to non-wasp sensitized AGS patients. α-Gal was not detected in wasp venom, but cross-reactivity between wasp and tick proteins was demonstrated which was not dependent on CCDs. The same cross-reactivity was also observed in the control sera. Furthermore, 17 putative cross-reactive peptides were identified using an in silico approach. Conclusions For the first time, cross-reactivity between wasp venom and tick proteins has been described. This may be a reason why the majority of Swedish AGS patients, who have all been tick bitten, are also sensitized against wasp

Supplementary data for article: Apostolovic, D.; Krstic, M.; Mihailovic, J.; Starkhammar, M.; Cirkovic Velickovic, T.; Hamsten, C.; Van Hage, M. Peptidomics of an in Vitro Digested α-Gal Carrying Protein Revealed IgE-Reactive Peptides. Scientific Reports 2017, 7 (1). https://doi.org/10.1038/s41598-017-05355-4

Supplementary material for: [https://doi.org/10.1038/s41598-017-05355-4]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2486

Th2-skewed T cells correlate with B cell response to α-Gal and tick antigens in α-Gal syndrome

Tick bites have been shown to transmit a novel form of severe food allergy, the galactose-α-1,3-galactose (α-Gal) syndrome (AGS). Cellular responses to α-Gal in patients with AGS have, to date, not been thoroughly scrutinized. Therefore, we investigated T and B cell proliferation, activation, and cytokine profiles in response to tick protein extract (TE) and α-Gal-free TE in patients with AGS and in healthy controls. T and B cells from both patients and controls proliferated in response to TE, but significantly more in patients with AGS. B cell proliferation, but not T cell proliferation, in patients with AGS was reduced by removing α-Gal from the TE. In addition, TE induced a clear Th2 cytokine profile in patients with AGS. Expression of CD23 by B cells correlated only to T cell proliferation. However, both B cell proliferation and CD23 expression were reduced when CD40L and IL-4 were blocked. A large portion of the IgG1 and IgE antibodies binding TE in patients with AGS were directed against the α-Gal epitope. We have, for what we believe to be the first time, investigated T and B cell responses to α-Gal carrying tick proteins in patients with AGS, which will be essential for the understanding of the immune response against an allergenic carbohydrate transmitted by ticks

Supplementary data for article: Apostolovic, D.; Tran, T. A. T.; Sánchez-Vidaurre, S.; Cirkovic Velickovic, T.; Starkhammar, M.; Hamsten, C.; Van Hage, M. Red Meat Allergic Patients Have a Selective IgE Response to the α-Gal Glycan. Allergy: European Journal of Allergy and Clinical Immunology 2015, 70 (11), 1497–1500. https://doi.org/10.1111/all.12695

Supporting information for: [https://doi.org/10.1111/all.12695]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1982