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The glycosylation pattern of common allergens: the recognition and uptake of Der p 1 by epithelial and dendritic cells is carbohydrate dependent.

Al-Ghouleh A, Johal R, Sharquie IK, Emara M, Harrington H, Shakib F, Ghaemmaghami AM - PLoS ONE (2012)

Bottom Line: Our data showed an increase in TSLP secretion by lung epithelia upon stimulation with natural Der p 1 which was carbohydrate dependent.The deglycosylated preparation of Der p 1 exhibited minimal uptake by DCs compared to the natural and hyperglycosylated recombinant counterparts, with the latter being taken up more readily than the other preparations.Collectively, our data indicate that carbohydrate moieties on allergens play a vital role in their recognition by innate immune cells, implicating them in downstream deleterious Th2 cell activation and IgE production.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Medical Sciences, Division of Immunology, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom.

ABSTRACT
Allergens are initiators of both innate and adaptive immune responses. They are recognised at the site of entry by epithelial and dendritic cells (DCs), both of which activate innate inflammatory circuits that can collectively induce Th2 immune responses. In an attempt to have a better understanding of the role of carbohydrates in the recognition and uptake of allergens by the innate immune system, we defined common glycosylation patterns in major allergens. This was done using labelled lectins and showed that allergens like Der p 1 (Dermatophagoides pteronyssinus group 1), Fel d 1 (Felis domisticus), Ara h 1 (Arachis hypogaea), Der p 2 (Dermatophagoides pteronyssinus group 2), Bla g 2 (Blattella germanica) and Can f 1 (Canis familiaris) are glycosylated and that the main dominant sugars on these allergens are 1-2, 1-3 and 1-6 mannose. These observations are in line with recent reports implicating the mannose receptor (MR) in allergen recognition and uptake by DCs and suggesting a major link between glycosylation and allergen recognition. We then looked at TSLP (Thymic Stromal Lymphopoietin) cytokine secretion by lung epithelia upon encountering natural Der p 1 allergen. TSLP is suggested to drive DC maturation in support of allergic hypersensitivity reactions. Our data showed an increase in TSLP secretion by lung epithelia upon stimulation with natural Der p 1 which was carbohydrate dependent. The deglycosylated preparation of Der p 1 exhibited minimal uptake by DCs compared to the natural and hyperglycosylated recombinant counterparts, with the latter being taken up more readily than the other preparations. Collectively, our data indicate that carbohydrate moieties on allergens play a vital role in their recognition by innate immune cells, implicating them in downstream deleterious Th2 cell activation and IgE production.

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A. Natural and recombinant Der p 1 uptake by immature DCs at 37°C compared to the non-allergen Staphopain B at the same conditions and concentrations. Results presented as MFI ± SDM and all preparations were labelled with FITC. B. Confocal images of the uptake of Staphopain B by immature DCs.
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pone-0033929-g004: A. Natural and recombinant Der p 1 uptake by immature DCs at 37°C compared to the non-allergen Staphopain B at the same conditions and concentrations. Results presented as MFI ± SDM and all preparations were labelled with FITC. B. Confocal images of the uptake of Staphopain B by immature DCs.

Mentions: Both natural and recombinant (hyperglycosylated) Der p 1 preparations are glycosylated albeit to different degrees. Staphopain B is also a cysteine protease protein, like Der p 1, but was shown to be an amannosylated antigen. To investigate the effect of glycosylation on the uptake of Der p 1 by DCs, we incubated all preparations under the same conditions with immature DCs at 37°C. All preparations were labelled by FITC and thus the uptake could be measured comparatively by flow cytometry as MFI readings. The control conditions for these experiments were DCs incubated with allergens at 4°C and DCs only. Levels of allergen uptake for each condition is presented as MFI (Fig. 2) and is also visualised using confocal imaging (Fig. 3). The results suggest that the average mean of uptake for recombinant Der p 1 (hyperglycosylated) is significantly (*P value<0.05) higher than that of natural Der p 1 at any given time point. We also studied the uptake of Staphopain B antigen, which is not known to induce any allergic reactions and is not mannosylated, and the results show minimal uptake of this non-allergen compared to Der p 1 (Fig. 4A & B).


The glycosylation pattern of common allergens: the recognition and uptake of Der p 1 by epithelial and dendritic cells is carbohydrate dependent.

Al-Ghouleh A, Johal R, Sharquie IK, Emara M, Harrington H, Shakib F, Ghaemmaghami AM - PLoS ONE (2012)

A. Natural and recombinant Der p 1 uptake by immature DCs at 37°C compared to the non-allergen Staphopain B at the same conditions and concentrations. Results presented as MFI ± SDM and all preparations were labelled with FITC. B. Confocal images of the uptake of Staphopain B by immature DCs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316510&req=5

pone-0033929-g004: A. Natural and recombinant Der p 1 uptake by immature DCs at 37°C compared to the non-allergen Staphopain B at the same conditions and concentrations. Results presented as MFI ± SDM and all preparations were labelled with FITC. B. Confocal images of the uptake of Staphopain B by immature DCs.
Mentions: Both natural and recombinant (hyperglycosylated) Der p 1 preparations are glycosylated albeit to different degrees. Staphopain B is also a cysteine protease protein, like Der p 1, but was shown to be an amannosylated antigen. To investigate the effect of glycosylation on the uptake of Der p 1 by DCs, we incubated all preparations under the same conditions with immature DCs at 37°C. All preparations were labelled by FITC and thus the uptake could be measured comparatively by flow cytometry as MFI readings. The control conditions for these experiments were DCs incubated with allergens at 4°C and DCs only. Levels of allergen uptake for each condition is presented as MFI (Fig. 2) and is also visualised using confocal imaging (Fig. 3). The results suggest that the average mean of uptake for recombinant Der p 1 (hyperglycosylated) is significantly (*P value<0.05) higher than that of natural Der p 1 at any given time point. We also studied the uptake of Staphopain B antigen, which is not known to induce any allergic reactions and is not mannosylated, and the results show minimal uptake of this non-allergen compared to Der p 1 (Fig. 4A & B).

Bottom Line: Our data showed an increase in TSLP secretion by lung epithelia upon stimulation with natural Der p 1 which was carbohydrate dependent.The deglycosylated preparation of Der p 1 exhibited minimal uptake by DCs compared to the natural and hyperglycosylated recombinant counterparts, with the latter being taken up more readily than the other preparations.Collectively, our data indicate that carbohydrate moieties on allergens play a vital role in their recognition by innate immune cells, implicating them in downstream deleterious Th2 cell activation and IgE production.

View Article: PubMed Central - PubMed

Affiliation: School of Molecular Medical Sciences, Division of Immunology, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom.

ABSTRACT
Allergens are initiators of both innate and adaptive immune responses. They are recognised at the site of entry by epithelial and dendritic cells (DCs), both of which activate innate inflammatory circuits that can collectively induce Th2 immune responses. In an attempt to have a better understanding of the role of carbohydrates in the recognition and uptake of allergens by the innate immune system, we defined common glycosylation patterns in major allergens. This was done using labelled lectins and showed that allergens like Der p 1 (Dermatophagoides pteronyssinus group 1), Fel d 1 (Felis domisticus), Ara h 1 (Arachis hypogaea), Der p 2 (Dermatophagoides pteronyssinus group 2), Bla g 2 (Blattella germanica) and Can f 1 (Canis familiaris) are glycosylated and that the main dominant sugars on these allergens are 1-2, 1-3 and 1-6 mannose. These observations are in line with recent reports implicating the mannose receptor (MR) in allergen recognition and uptake by DCs and suggesting a major link between glycosylation and allergen recognition. We then looked at TSLP (Thymic Stromal Lymphopoietin) cytokine secretion by lung epithelia upon encountering natural Der p 1 allergen. TSLP is suggested to drive DC maturation in support of allergic hypersensitivity reactions. Our data showed an increase in TSLP secretion by lung epithelia upon stimulation with natural Der p 1 which was carbohydrate dependent. The deglycosylated preparation of Der p 1 exhibited minimal uptake by DCs compared to the natural and hyperglycosylated recombinant counterparts, with the latter being taken up more readily than the other preparations. Collectively, our data indicate that carbohydrate moieties on allergens play a vital role in their recognition by innate immune cells, implicating them in downstream deleterious Th2 cell activation and IgE production.

Show MeSH
Related in: MedlinePlus